Ai6

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相关技术服务:质粒构建    基因合成    质粒大提

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载体基本信息

质粒名称: Ai6 
出品公司: Addgene
目录编号: 22798
存储实验室: Hongkui Zeng 

载体骨架基本信息
载体名称: Rosa26 Targeting Vector 
载体出品公司: --
载体抗性: 氨苄青霉素
空载体大小: 9862 bp
完整质粒大小: --
载体修饰: --
载体类型: Mouse Targeting
筛选标记: 新霉素G418
高拷贝/低拷贝: 高拷贝
生长条件: 37 ℃

插入基因信息
插入基因名称: CAG-Floxed ZsGreen
物种来源: n/a
插入大小: 6319 bp
基因突变: --
GenBank编号 --
启动子 --
标签/融合蛋白: --
克隆方法: 限制性内切酶
5' 克隆位点: PacI(未破坏)
3' 克隆位点: AscI (未破坏)
5' 测序引物 n/a
3' 测序引物 n/a

文献引用方法
材料和方法部分: Ai6 was a gift from Hongkui Zeng (Addgene plasmid # 22798 ; http://n2t.net/addgene:22798 ; RRID:Addgene_22798)
参考文献部分: A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Madisen L, Zwingman TA, Sunkin SM, Oh SW, Zariwala HA, Gu H, Ng LL, Palmiter RD, Hawrylycz MJ, Jones AR, Lein ES, Zeng H. Nat Neurosci. 2010 Jan . 13(1):133-40. 10.1038/nn.2467 PubMed 20023653

载体图谱质粒图谱和多克隆位点信息

Ai6质粒图谱

载体简介

载体序列

LOCUS       Ai6              16015 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Ai6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16015)
  AUTHORS   Madisen L, Zwingman TA, Sunkin SM, Oh SW, Zariwala HA, Gu H, Ng LL, 
            Palmiter RD, Hawrylycz MJ, Jones AR, Lein ES, Zeng H
  TITLE     A robust and high-throughput Cre reporting and characterization 
            system for the whole mouse brain.
  JOURNAL   Nat Neurosci. 2010 Jan . 13(1):133-40.
  PUBMED    20023653
REFERENCE   2  (bases 1 to 16015)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..16015
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        947..1250
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1252..1529
                     /label=chicken beta-actin promoter
     intron          1530..2546
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2470..2492
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2554..2573
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    2622..2655
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(2679..2712)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     polyA_signal    2879..3013
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3133..3267
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    3387..3521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(3550..3583)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      3644..3653
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3650..4345
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein"
                     /label=ZsGreen1
                     /note="mammalian codon-optimized"
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     misc_feature    4393..4981
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4446..4466)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4864..4875)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4984..5000)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4985..5001)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    5008..5215
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    5223..5256
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        5278..5777
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     protein_bind    5797..5830
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      5831..5840
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             5837..6640
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5900..5919)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6510..6529
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6650..7111
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     misc_feature    7177..11517
                     /label=Rosa26 right arm
                     /note="right homology arm from intron 1 of the mouse Rosa26
                     gene (Soriano, 1999)"
     promoter        11567..12066
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             12093..12665
                     /codon_start=1
                     /gene="Corynebacterium diphtheriae tox"
                     /product="diphtheria toxin fragment A"
                     /label=DTA
                     /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD
                     DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL
                     GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI
                     NFETRGKRGQDAMYEYMAQACAGNRVRR"
     intron          12742..12807
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     polyA_signal    12881..13105
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(13167..13187)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(13167..13185)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(13206..13222)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13206..13222)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13219..13241)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13230..13246
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(13254..13284)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13299..13320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(13437..13454)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(13608..14196)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(13688..13707)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(14367..15227)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     14990..15009
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15228..15332)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      15359..15814
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(15446..15465)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     15656..15677
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     15940..15962
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     15954..15971
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     15955..15971
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     15981..16000
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        15981..15999
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ccgcggcagg ccctccgagc gtggtggagc cgttctgtga gacagccggg tacgagtcgt
       61 gacgctggaa ggggcaagcg ggtggtgggc aggaatgcgg tccgccctgc agcaaccgga
      121 gggggaggga gaagggagcg gaaaagtctc caccggacgc ggccatggct cggggggggg
      181 gggggggagg gagggtcagc gaaagtagct cgcgcgcgag cggccgccca ccctcccctt
      241 cctctggggg agtcgtttta cccgccgccg gccgggcctc gtcgtctgat tggctctcgg
      301 ggcccagaaa actggccctt gccattggct cgtgttcgtg caagttgagt ccatccgccg
      361 gccagcgggg gcggcgagga ggcgctccca ggttccggcc ctcccctcgg ccccgcgccg
      421 cagagtctgg ccgcgcgccc ctgcgcaacg tggcaggaag cgcgcgctgg gggcggggac
      481 gggcagtagg gctgagcggc tgcggggcgg gtgcaagcac gtttccgact tgagttgcct
      541 caagaggggc gtgctgagcc agacctccat cgcgcactcc ggggagtgga gggaaggagc
      601 gagggctcag ttgggctgtt ttggaggcag gaagcacttg ctctcccaaa gtcgctctga
      661 gttgttatca gtaagggagc tgcagtggag taggcgggga gaaggccgca cccttctccg
      721 gaggggggag gggagtgttg caataccttt ctgggagttc tctgctgcct cctggcttct
      781 gaggaccgcc ctgggcctgg gagaatccct tccccctctt ccctcgtgat ctgcaactcc
      841 agtctttcta gccttaatta accgtttaaa caattctgca ggaatctagt tattaatagt
      901 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      961 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     1021 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     1081 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     1141 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     1201 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga
     1261 gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt
     1321 tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc
     1381 caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag
     1441 ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc
     1501 cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc
     1561 cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac
     1621 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac
     1681 ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt
     1741 gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg
     1801 gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc
     1861 gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg
     1921 aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg
     1981 cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag cacggcccgg
     2041 cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg ggcggggggt
     2101 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg
     2161 aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt
     2221 gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa atctgtgcgg
     2281 agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg
     2341 gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc
     2401 ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg
     2461 gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg
     2521 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc
     2581 atcattttgg caaagaattg atttgatacc gcgggcccta agaagttcct attctctaga
     2641 aagtatagga acttcgtcga catttaaatc atttaaatat aacttcgtat aatgtatgct
     2701 atacgaagtt attcgcgatg aataaatgaa agcttgcaga tctgcgactc tagaggatct
     2761 gcgactctag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     2821 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     2881 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2941 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3001 tcatgtctgg atctgcgact ctagaggatc ataatcagcc ataccacatt tgtagaggtt
     3061 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     3121 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     3181 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     3241 atcaatgtat cttatcatgt ctggatctgc gactctagag gatcataatc agccatacca
     3301 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3361 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3421 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3481 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccccatcaag ctgatccgga
     3541 acccttaata taacttcgta taatgtatgc tatacgaagt tattaggtcc ctcgacctgc
     3601 agcccaagct agatcgaatt cggccggccg atatcggcgc gccgccacca tggcccagtc
     3661 caagcacggc ctgaccaagg agatgaccat gaagtaccgc atggagggct gcgtggacgg
     3721 ccacaagttc gtgatcaccg gcgagggcat cggctacccc ttcaagggca agcaggccat
     3781 caacctgtgc gtggtggagg gcggcccctt gcccttcgcc gaggacatct tgtccgccgc
     3841 cttcatgtac ggcaaccgcg tgttcaccga gtacccccag gacatcgtcg actacttcaa
     3901 gaactcctgc cccgccggct acacctggga ccgctccttc ctgttcgagg acggcgccgt
     3961 gtgcatctgc aacgccgaca tcaccgtgag cgtggaggag aactgcatgt accacgagtc
     4021 caagttctac ggcgtgaact tccccgccga cggccccgtg atgaagaaga tgaccgacaa
     4081 ctgggagccc tcctgcgaga agatcatccc cgtgcccaag cagggcatct tgaagggcga
     4141 cgtgagcatg tacctgctgc tgaaggacgg tggccgcttg cgctgccagt tcgacaccgt
     4201 gtacaaggcc aagtccgtgc cccgcaagat gcccgactgg cacttcatcc agcacaagct
     4261 gacccgcgag gaccgcagcg acgccaagaa ccagaagtgg cacctgaccg agcacgccat
     4321 cgcctccggc tccgccttgc cctgataaga tatcggccgg ccctgcagga attcgatatc
     4381 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     4441 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     4501 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     4561 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     4621 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4681 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4741 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     4801 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4861 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4921 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4981 catcgatacc gtcgacctcg acctcgactg tgccttctag ttgccagcca tctgttgttt
     5041 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     5101 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     5161 tggggcagga cagcaagggg gaggattggg aagacaatgg caggcatgct ggggaactag
     5221 tggtgccagg gcgtgccctt gggctccccg ggcgcggcgg ccgcatcgaa ttctaccggg
     5281 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca
     5341 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc
     5401 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga
     5461 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc
     5521 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg
     5581 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg
     5641 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg
     5701 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct
     5761 catctccggg cctttcgacc tgcaatcgcc gctagcgaag ttcctattct ctagaaagta
     5821 taggaacttc gccaccatgg gatcggccat tgaacaagat ggattgcacg caggttctcc
     5881 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     5941 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     6001 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     6061 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     6121 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     6181 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     6241 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     6301 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     6361 caggctcaag gcgcgcatgc ccgacggcga tgatctcgtc gtgacccatg gcgatgcctg
     6421 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     6481 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     6541 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6601 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ggggatccgc tgtaagtctg
     6661 cagaaattga tgatctatta aacaataaag atgtccacta aaatggaagt ttttcctgtc
     6721 atactttgtt aagaagggtg agaacagagt acctacattt tgaatggaag gattggagct
     6781 acgggggtgg gggtggggtg ggattagata aatgcctgct ctttactgaa ggctctttac
     6841 tattgcttta tgataatgtt tcatagttgg atatcataat ttaaacaagc aaaaccaaat
     6901 taagggccag ctcattcctc ccactcatga tctatagatc tatagatctc tcgtgggatc
     6961 attgtttttc tcttgattcc cactttgtgg ttctaagtac tgtggtttcc aaatgtgtca
     7021 gtttcatagc ctgaagaacg agatcagcag cctctgttcc acatacactt cattctcagt
     7081 attgttttgc caagttctaa ttccatcaga aagcttgcag aagatctccc caactggggt
     7141 aacctttgag ttctctcagt tgggggggcg cgccggctag aagatgggcg ggagtcttct
     7201 gggcaggctt aaaggctaac ctggtgtgtg ggcgttgtcc tgcaggggaa ttgaacaggt
     7261 gtaaaattgg agggacaaga cttcccacag attttcggtt ttgtcgggaa gttttttaat
     7321 aggggcaaat aaggaaaatg ggaggatagg tagtcatctg gggttttatg cagcaaaact
     7381 acaggttatt attgcttgtg atccgcctcg gagtattttc catcgaggta gattaaagac
     7441 atgctcaccc gagttttata ctctcctgct tgagatcctt actacagtat gaaattacag
     7501 tgtcgcgagt tagactatgt aagcagaatt ttaatcattt ttaaagagcc cagtacttca
     7561 tatccatttc tcccgctcct tctgcagcct tatcaaaagg tattttagaa cactcatttt
     7621 agccccattt tcatttatta tactggctta tccaacccct agacagagca ttggcatttt
     7681 ccctttcctg atcttagaag tctgatgact catgaaacca gacagattag ttacatacac
     7741 cacaaatcga ggctgtagct ggggcctcaa cactgcagtt cttttataac tccttagtac
     7801 actttttgtt gatcctttgc cttgatcctt aattttcagt gtctatcacc tctcccgtca
     7861 ggtggtgttc cacatttggg cctattctca gtccagggag ttttacaaca atagatgtat
     7921 tgagaatcca acctaaagct taactttcca ctcccatgaa tgcctctctc ctttttctcc
     7981 atttataaac tgagctatta accattaatg gtttccaggt ggatgtctcc tcccccaata
     8041 ttacctgatg tatcttacat attgccaggc tgatatttta agacattaaa aggtatattt
     8101 cattattgag ccacatggta ttgattactg cttactaaaa ttttgtcatt gtacacatct
     8161 gtaaaaggtg gttccttttg gaatgcaaag ttcaggtgtt tgttgtcttt cctgacctaa
     8221 ggtcttgtga gcttgtattt tttctattta agcagtgctt tctcttggac tggcttgact
     8281 catggcattc tacacgttat tgctggtcta aatgtgattt tgccaagctt cttcaggacc
     8341 tataattttg cttgacttgt agccaaacac aagtaaaatg attaagcaac aaatgtattt
     8401 gtgaagcttg gtttttaggt tgttgtgttg tgtgtgcttg tgctctataa taatactatc
     8461 caggggctgg agaggtggct cggagttcaa gagcacagac tgctcttcca gaagtcctga
     8521 gttcaattcc cagcaaccac atggtggctc acaaccatct gtaatgggat ctgatgccct
     8581 cttctggtgt gtctgaagac cacaagtgta ttcacattaa ataaataaat cctccttctt
     8641 cttctttttt ttttttttaa agagaatact gtctccagta gaatttactg aagtaatgaa
     8701 atactttgtg tttgttccaa tatggtagcc aataatcaaa ttactcttta agcactggaa
     8761 atgttaccaa ggaactaatt tttatttgaa gtgtaactgt ggacagagga gccataactg
     8821 cagacttgtg ggatacagaa gaccaatgca gactttaatg tcttttctct tacactaagc
     8881 aataaagaaa taaaaattga acttctagta tcctatttgt ttaaactgct agctttactt
     8941 aacttttgtg cttcatctat acaaagctga aagctaagtc tgcagccatt actaaacatg
     9001 aaagcaagta atgataattt tggatttcaa aaatgtaggg ccagagttta gccagccagt
     9061 ggtggtgctt gcctttatgc ctttaatccc agcactctgg aggcagagac aggcagatct
     9121 ctgagtttga gcccagcctg gtctacacat caagttctat ctaggatagc caggaataca
     9181 cacagaaacc ctgttgggga ggggggctct gagatttcat aaaattataa ttgaagcatt
     9241 ccctaatgag ccactatgga tgtggctaaa tccgtctacc tttctgatga gatttgggta
     9301 ttattttttc tgtctctgct gttggttggg tcttttgaca ctgtgggctt tctttaaagc
     9361 ctccttcctg ccatgtggtc tcttgtttgc tactaacttc ccatggctta aatggcatgg
     9421 ctttttgcct tctaagggca gctgctgaga tttgcagcct gatttccagg gtggggttgg
     9481 gaaatctttc aaacactaaa attgtccttt aatttttttt ttaaaaaatg ggttatataa
     9541 taaacctcat aaaatagtta tgaggagtga ggtggactaa tattaaatga gtccctcccc
     9601 tataaaagag ctattaaggc tttttgtctt atacttaact ttttttttaa atgtggtatc
     9661 tttagaacca agggtcttag agttttagta tacagaaact gttgcatcgc ttaatcagat
     9721 tttctagttt caaatccaga gaatccaaat tcttcacagc caaagtcaaa ttaagaattt
     9781 ctgactttta atgttaattt gcttactgtg aatataaaaa tgatagcttt tcctgaggca
     9841 gggtctcact atgtatctct gcctgatctg caacaagata tgtagactaa agttctgcct
     9901 gcttttgtct cctgaatact aaggttaaaa tgtagtaata cttttggaac ttgcaggtca
     9961 gattctttta taggggacac actaagggag cttgggtgat agttggtaaa atgtgtttca
    10021 agtgatgaaa acttgaatta ttatcaccgc aacctacttt ttaaaaaaaa aagccaggcc
    10081 tgttagagca tgcttaaggg atccctagga cttgctgagc acacaagagt agttacttgg
    10141 caggctcctg gtgagagcat atttcaaaaa acaaggcaga caaccaagaa actacagtta
    10201 aggttacctg tctttaaacc atctgcatat acacagggat attaaaatat tccaaataat
    10261 atttcattca agttttcccc catcaaattg ggacatggat ttctccggtg aataggcaga
    10321 gttggaaact aaacaaatgt tggttttgtg atttgtgaaa ttgttttcaa gtgatagtta
    10381 aagcccatga gatacagaac aaagctgcta tttcgaggtc tcttggttta tactcagaag
    10441 cacttctttg ggtttccctg cactatcctg atcatgtgct aggcctacct taggctgatt
    10501 gttgttcaaa taaacttaag tttcctgtca ggtgatgtca tatgatttca tatatcaagg
    10561 caaaacatgt tatatatgtt aaacatttgt acttaatgtg aaagttaggt ctttgtgggt
    10621 ttgattttta attttcaaaa cctgagctaa ataagtcatt tttacatgtc ttacatttgg
    10681 tggaattgta taattgtggt ttgcaggcaa gactctctga cctagtaacc ctacctatag
    10741 agcactttgc tgggtcacaa gtctaggagt caagcatttc accttgaagt tgagacgttt
    10801 tgttagtgta tactagttta tatgttggag gacatgttta tccagaagat attcaggact
    10861 atttttgact gggctaagga attgattctg attagcactg ttagtgagca ttgagtggcc
    10921 tttaggcttg aattggagtc acttgtatat ctcaaataat gctggccttt tttaaaaagc
    10981 ccttgttctt tatcaccctg ttttctacat aatttttgtt caaagaaata cttgtttgga
    11041 tctccttttg acaacaatag catgttttca agccatattt tttttccttt tttttttttt
    11101 ttttggtttt tcgagacagg gtttctctgt atagccctgg ctgtcctgga actcactttg
    11161 tagaccaggc tggcctcgaa ctcagaaatc cgcctgcctc tgcctcctga gtgccgggat
    11221 taaaggcgtg caccaccacg cctggctaag ttggatattt tgttatataa ctataaccaa
    11281 tactaactcc actgggtgga tttttaattc agtcagtagt cttaagtggt ctttattggc
    11341 ccttcattaa aatctactgt tcactctaac agaggctgtt ggtactagtg gcacttaagc
    11401 aacttcctac ggatatacta gcagattaag ggtcagggat agaaactagt ctagcgtttt
    11461 gtatacctac cagctttata ctaccttgtt ctgatagaaa tatttcagga catctagctt
    11521 atcgataccg tcgacggtat cgataagctt gatatcgaat tctaccgggt aggggaggcg
    11581 cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac
    11641 acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg
    11701 ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc
    11761 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc
    11821 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa
    11881 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc
    11941 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca
    12001 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc
    12061 ctttcgacct gcaggtcctc gccatggatc ctgatgatgt tgttgattct tctaaatctt
    12121 ttgtgatgga aaacttttct tcgtaccacg ggactaaacc tggttatgta gattccattc
    12181 aaaaaggtat acaaaagcca aaatctggta cacaaggaaa ttatgacgat gattggaaag
    12241 ggttttatag taccgacaat aaatacgacg ctgcgggata ctctgtagat aatgaaaacc
    12301 cgctctctgg aaaagctgga ggcgtggtca aagtgacgta tccaggactg acgaaggttc
    12361 tcgcactaaa agtggataat gccgaaacta ttaagaaaga gttaggttta agtctcactg
    12421 aaccgttgat ggagcaagtc ggaacggaag agtttatcaa aaggttcggt gatggtgctt
    12481 cgcgtgtagt gctcagcctt cccttcgctg aggggagttc tagcgttgaa tatattaata
    12541 actgggaaca ggcgaaagcg ttaagcgtag aacttgagat taattttgaa acccgtggaa
    12601 aacgtggcca agatgcgatg tatgagtata tggctcaagc ctgtgcagga aatcgtgtca
    12661 ggcgatctct ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct
    12721 acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta
    12781 ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag
    12841 tggtggaatg cagatcctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
    12901 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
    12961 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
    13021 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
    13081 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgagg
    13141 gggggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
    13201 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
    13261 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
    13321 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
    13381 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
    13441 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
    13501 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
    13561 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
    13621 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
    13681 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
    13741 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
    13801 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
    13861 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
    13921 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
    13981 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
    14041 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
    14101 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
    14161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
    14221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
    14281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
    14341 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
    14401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
    14461 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
    14521 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
    14581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
    14641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
    14701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
    14761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
    14821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
    14881 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
    14941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
    15001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
    15061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
    15121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
    15181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
    15241 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
    15301 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
    15361 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
    15421 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
    15481 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
    15541 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
    15601 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
    15661 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
    15721 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
    15781 gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact
    15841 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
    15901 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
    15961 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattggag ctcca
//

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