pAAVS1-P-CAG-GFP

价格:1200元

联系方式:I47-825O-882O

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: pAAVS1-P-CAG-GFP 
出品公司: 爱迪基因
目录编号: 80491
存储实验室: Knut Woltjen

载体骨架基本信息
载体名称: pCR-TOPO
载体出品公司: --
载体抗性: 氨苄青霉素卡那霉素
空载体大小: --
完整质粒大小: 3000 bp
载体修饰: --
载体类型: 哺乳细胞表达
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-GFP
物种来源: --
插入大小: 2447 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: AgeI(未破坏)
3' 克隆位点: NotI (未破坏)
5' 测序引物 CMV-F
3' 测序引物 SV40pA-R

文献引用方法
材料和方法部分: pAAVS1-P-CAG-GFP was a gift from Knut Woltjen (Addgene plasmid # 80491 ; http://n2t.net/addgene:80491 ; RRID:Addgene_80491)
参考文献部分: A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. Nodvig CS, Nielsen JB, Kogle ME, Mortensen UH. PLoS One. 2015 Jul 15;10(7):e0133085. doi: 10.1371/journal.pone.0133085. eCollection 2015. PONE-D-15-11561 [pii] PubMed 26177455

载体图谱质粒图谱和多克隆位点信息

pAAVS1-P-CAG-GFP质粒图谱

载体简介

载体序列

LOCUS       pAAVS1-P-CAG-GFP                9608 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  donor vector for AAVS1 targeting (puromycin selection) and 
            constitutive GFP expression.
ACCESSION   .
VERSION     .
KEYWORDS    pAAVS1-P-CAG-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9608)
  AUTHORS   Oceguera-Yanez F, Kim SI, Matsumoto T, Tan GW, Xiang L, Hatani T, 
            Kondo T, Ikeya M, Yoshida Y, Inoue H, Woltjen K
  TITLE     Engineering the AAVS1 locus for consistent and scalable transgene 
            expression in human iPSCs and their differentiated derivatives.
  JOURNAL   Methods. 2015 Dec 18. pii: S1046-2023(15)30181-X. doi: 
            10.1016/j.ymeth.2015.12.012.
  PUBMED    26707206
REFERENCE   2  (bases 1 to 9608)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9608
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        242..260
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    295..1098
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    1105..1130
                     /label=SA
                     /note="splice acceptor site"
     CDS             1154..1207
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             1217..1816
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1217..1236)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1713..1733
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(1848..1865)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1854..2078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        2082..2461
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2464..2739
                     /label=chicken beta-actin promoter
     intron          2740..3748
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3672..3694
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3756..3775
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3803..3812
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3809..4528
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3854..3875)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4115..4134)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4462..4483
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4612..4631)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_site      4677..4732
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(4731..4750)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     misc_feature    5104..5940
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(5980..5999)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5981..5999)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6006..6023)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6006..6022)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6015..6037)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             6160..6462
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     6341..6360
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             6811..7605
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6865..6884)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7475..7494
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             complement(7855..8715)
                     /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     8478..8497
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8839..9427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9328..9347
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9581..9598
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca
      241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc cctttgcttt
      301 ctctgaccag cattctctcc cctgggcctg tgccgctttc tgtctgcagc ttgtggcctg
      361 ggtcacctct acggctggcc cagatccttc cctgccgcct ccttcaggtt ccgtcttcct
      421 ccactccctc ttccccttgc tctctgctgt gttgctgccc aaggatgctc tttccggagc
      481 acttccttct cggcgctgca ccacgtgatg tcctctgagc ggatcctccc cgtgtctggg
      541 tcctctccgg gcatctctcc tccctcaccc aaccccatgc cgtcttcact cgctgggttc
      601 ccttttcctt ctccttctgg ggcctgtgcc atctctcgtt tcttaggatg gccttctccg
      661 acggatgtct cccttgcgtc ccgcctcccc ttcttgtagg cctgcatcat caccgttttt
      721 ctggacaacc ccaaagtacc ccgtctccct ggctttagcc acctctccat cctcttgctt
      781 tctttgcctg gacaccccgt tctcctgtgg attcgggtca cctctcactc ctttcatttg
      841 ggcagctccc ctacccccct tacctctcta gtctgtgcta gctcttccag ccccctgtca
      901 tggcatcttc caggggtccg agagctcagc tagtcttctt cctccaaccc gggcccctat
      961 gtccacttca ggacagcatg tttgctgcct ccagggatcc tgtgtccccg agctgggacc
     1021 accttatatt cccagggccg gttaatgtgg ctctggttct gggtactttt atctgtcccc
     1081 tccaccccac agtggggcaa gcttctgacc tcttctcttc ctcccacagg gcctcgagag
     1141 atctggcagc ggagagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc
     1201 cggccctagg ctcgagatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     1261 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     1321 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     1381 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     1441 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     1501 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     1561 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     1621 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     1681 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     1741 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     1801 caagcccggt gcctgatcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc
     1861 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1921 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     1981 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     2041 caatagcagg catgctgggg atgcggtggg ctctatgggt cgacattgat tattgactag
     2101 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     2161 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     2221 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     2281 ggtggactat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     2341 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     2401 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     2461 gggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc
     2521 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg
     2581 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt
     2641 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
     2701 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gttgccttcg
     2761 ccccgtgccc cgctccgcgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
     2821 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
     2881 taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc
     2941 cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc
     3001 cgcgtgcggc ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt
     3061 gcgctccgcg tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg
     3121 ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt
     3181 gggcgcggcg gtcgggctgt aacccccccc tgcacccccc tccccgagtt gctgagcacg
     3241 gcccggcttc gggtgcgggg ctccgtgcgg ggcgtggcgc ggggctcgcc gtgccgggcg
     3301 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct
     3361 cgggggaggg gcgcggcggc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag
     3421 ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg
     3481 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg gcgaagcggt
     3541 gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc
     3601 cccttctcca tctccagcct cggggctgcc gcagggggac ggctgccttc gggggggacg
     3661 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
     3721 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc
     3781 tcatcatttt ggcaaagaat tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg
     3841 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
     3901 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
     3961 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
     4021 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
     4081 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
     4141 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
     4201 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
     4261 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
     4321 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
     4381 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
     4441 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     4501 tctcggcatg gacgagctgt acaagtaaga attcactcct caggtgcagg ctgcctatca
     4561 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc
     4621 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata
     4681 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag
     4741 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc
     4801 aacatatgcc atatgctggc tgccatgaac aaaggtggct ataaagaggt catcagtata
     4861 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     4921 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     4981 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     5041 ttctcttatg aagatccctc gacctgcagc ccaagcttgg atccctcgac agtactaagc
     5101 tttactaggg acaggattgg tgacagaaaa gccccatcct taggcctcct ccttcctagt
     5161 ctcctgatat tgggtctaac ccccacctcc tgttaggcag attccttatc tggtgacaca
     5221 cccccatttc ctggagccat ctctctcctt gccagaacct ctaaggtttg cttacgatgg
     5281 agccagagag gatcctggga gggagagctt ggcagggggt gggagggaag ggggggatgc
     5341 gtgacctgcc cggttctcag tggccaccct gcgctaccct ctcccagaac ctgagctgct
     5401 ctgacgcggc tgtctggtgc gtttcactga tcctggtgct gcagcttcct tacacttccc
     5461 aagaggagaa gcagtttgga aaaacaaaat cagaataagt tggtcctgag ttctaacttt
     5521 ggctcttcac ctttctagtc cccaatttat attgttcctc cgtgcgtcag ttttacctgt
     5581 gagataaggc cagtagccag ccccgtcctg gcagggctgt ggtgaggagg ggggtgtccg
     5641 tgtggaaaac tccctttgtg agaatggtgc gtcctaggtg ttcaccaggt cgtggccgcc
     5701 tctactccct ttctctttct ccatccttct ttccttaaag agtccccagt gctatctggg
     5761 acatattcct ccgcccagag cagggtcccg cttccctaag gccctgctct gggcttctgg
     5821 gtttgagtcc ttggcaagcc caggagaggc gctcaggctt ccctgtcccc cttcctcgtc
     5881 caccatctca tgcccctggc tctcctgccc cttccctaca ggggttcctg gctctgctct
     5941 aagggcaagg gcgaattcgc ggccgctaaa ttcaattcgc cctatagtga gtcgtattac
     6001 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     6061 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     6121 gatcgccctt cccaacagtt gcgcagccta tacgtacggc agtttaaggt ttacacctat
     6181 aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccg
     6241 gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt
     6301 gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg
     6361 gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat
     6421 gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg catgagatta
     6481 tcaaaaagga tcttcaccta gatccttttc acgtagaaag ccagtccgca gaaacggtgc
     6541 tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag
     6601 agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg
     6661 acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc
     6721 aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagctct
     6781 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     6841 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     6901 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     6961 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     7021 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     7081 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     7141 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     7201 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     7261 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     7321 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     7381 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     7441 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     7501 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     7561 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaattat taacgcttac
     7621 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatcagg
     7681 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     7741 aaatatgtat ccgctcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7801 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7861 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7921 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7981 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     8041 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     8101 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     8161 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     8221 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     8281 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8341 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8401 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8461 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8521 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8581 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8641 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8701 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8761 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     8821 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     8881 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     8941 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     9001 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     9061 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     9121 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     9181 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     9241 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     9301 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     9361 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     9421 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     9481 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     9541 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     9601 agcggaag
//

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