pcDNA5-T/O-Ds1-mCherry

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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: pcDNA5-T/O-Ds1-mCherry
出品公司: Addgene
目录编号: 112855
存储实验室: David Sprinzak

载体骨架基本信息
载体名称: pcDNA5/TO
载体出品公司: Thermofisher
载体类型: 哺乳细胞表达
筛选标记: 潮霉素
载体抗性: 氨苄霉素
生长温度: 37 ℃
宿主菌: DH5a
拷贝数: 高拷贝

插入基因信息
插入基因名称: DCHS1
基因又名: dachsous cadherin-related 1, DCHS1 (a.k.a. CDH19, CDH25, CDHR6, FIB1, MVP2, PCDH16, VMLDS1)
物种来源: H. sapiens (human)
启动子: doxycycline inducible promoter
标签/融合蛋白: mCherry (C terminal on insert)
克隆方法: 限制性内切酶
5’克隆位点: NotI(未破坏)
3’克隆位点: AgeI (未破坏)
5’测序引物 CMV-F

文献引用方法
材料和方法部分: pcDNA5-T/O-Ds1-mCherry was a gift from David Sprinzak (Addgene plasmid # 112855 ; http://n2t.net/addgene:112855 ; RRID:Addgene_112855)
参考文献部分: A synthetic planar cell polarity system reveals localized feedback on Fat4-Ds1 complexes. Loza O, Heemskerk I, Gordon-Bar N, Amir-Zilberstein L, Jung Y, Sprinzak D. Elife. 2017 Aug 18;6. pii: 24820. doi: 10.7554/eLife.24820. 24820 [pii] PubMed 28826487

载体图谱质粒图谱和多克隆位点信息

pcDNA5-TO-Ds1-mCherry载体图谱

载体简介

载体序列

LOCUS       pcDNA5-T/O-Ds1-mCherry               16256 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Dox inducible expression in mammalian cells of Ds1 protein fused to 
            mCherry fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA5-T/O-Ds1-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 16256)
  AUTHORS   Loza O, Heemskerk I, Gordon-Bar N, Amir-Zilberstein L, Jung Y, 
            Sprinzak D
  TITLE     A synthetic planar cell polarity system reveals localized feedback 
            on Fat4-Ds1 complexes.
  JOURNAL   Elife. 2017 Aug 18;6. pii: 24820. doi: 10.7554/eLife.24820.
  PUBMED    28826487
REFERENCE   2  (bases 1 to 16256)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..16256
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             10693..11406
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYKS"
     primer_bind     complement(10838..10856)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     11106..11125
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(11444..11461)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    11450..11674
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11720..12148
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(11807..11826)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     12017..12038
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(12157..12177)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        12162..12491
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      12342..12477
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     12404..12423
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             12540..13565
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    13695..13816
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(13732..13751)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     13786..13805
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(13865..13881)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(13865..13881)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(13878..13900)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    13889..13905
                     /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(13913..13943)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    13958..13979
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(14096..14113)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(14267..14855)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(14347..14366)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(15026..15886)
                     /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     15649..15668
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(15887..15991)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(16066..16085)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccgcg gccgctcgcc
      781 accatgcaga aggagctggg cattgtgcct tcctgccctg gcatgaagag ccccaggccc
      841 cacctcctgc taccattgct gctgctgctg ctgctgctgc tgggggctgg ggtgccaggt
      901 gcctggggtc aggctgggag cctggacttg cagattgatg aggagcagcc agcgggtaca
      961 ctgattggcg acatcagtgc ggggcttccg gcaggcacgg cagctcctct catgtacttc
     1021 atctctgccc aagagggcag cggcgtgggc acagacctgg ccattgacga acacagtggg
     1081 gtcgtccgta cagcccgtgt cttggaccgt gagcagcggg accgctaccg cttcactgca
     1141 gtcactcctg atggtgccac cgtagaagtt acagtgcgag tggctgacat caacgaccat
     1201 gctccagcct tcccacaggc tcgggctgcc ctgcaggtac ctgagcatac agcttttggc
     1261 acccgctacc cactggagcc tgctcgtgat gcagatgctg ggcgtctggg aacccagggc
     1321 tatgcgctat ctggtgatgg ggctggagag accttccggc tggagacacg ccccggtcca
     1381 gatgggactc cagtacctga gctggtagtt actggggaac tggaccgaga gaaccgctca
     1441 cactatatgc tacagctgga ggcctatgat ggtggttcac ccccccggag ggcccaggcc
     1501 ctgctggacg tgacactgct ggacatcaat gaccatgccc cggctttcaa tcagagccgc
     1561 taccatgctg tggtgtctga gagcctggcc cctggcagtc ctgtcttgca ggtgttcgca
     1621 tctgatgccg atgctggtgt caatggggct gtgacttacg agatcaaccg gaggcagagc
     1681 gagggtgatg gacccttctc catcgacgca cacacggggc tgctgcagtt agagcggcca
     1741 ctggactttg agcagcggcg ggtccatgaa ctggtggtgc aagcacgaga tggtggggct
     1801 caccctgagc tgggctcggc ctttgtgact gtgcatgtgc gagatgccaa tgacaatcag
     1861 ccctccatga ctgtcatctt tctcagtgca gatggctccc cccaagtgtc tgaggccgcc
     1921 ccacctggac agctcgttgc tcgcatctct gtgtcagacc cagatgatgg tgactttgcc
     1981 catgtcaatg tgtccctgga aggtggagag ggccactttg ccctaagcac ccaagacagc
     2041 gtcatctatc tggtgtgtgt ggctcggcgg ctggatcgag aggagaggga tgcctataac
     2101 ttgagggtta cagccacaga ctcaggctca cctccactgc gggctgaggc tgcctttgtg
     2161 ctgcacgtca ctgatgtcaa cgacaatgca cctgcctttg accgccagct ctaccgacct
     2221 gagcccctgc ctgaggttgc gctgcctggc agctttgtag tgcgggtgac tgctcgggat
     2281 cctgaccaag gcaccaatgg tcaggtcact tatagcctag cccctggcgc ccacacccac
     2341 tggttctcca ttgaccccac ctcaggcatt atcactacgg ctgcctcact ggactatgag
     2401 ttggaacctc agccacagct gattgtggtg gccacagatg gtggcctgcc ccctctagcc
     2461 tcctctgcca cagttagcgt ggccctgcaa gatgtgaatg ataatgagcc ccaattccag
     2521 aggactttct acaatgcctc actgcctgag ggcacccagc ctggaacttg cttcctgcag
     2581 gtgacagcca cagacgcgga tagtggccca tttggcctcc tctcctattc cttgggtgct
     2641 ggacttgggt cctccggatc tcccccattc cgcattgatg cccatagcgg tgatgtgtgc
     2701 acaacccgga ccctggaccg tgaccagggg ccctcaagct ttgacttcac agtgacagct
     2761 gtggatgggg gaggcctcaa gtccatggta tatgtgaagg tgtttctgtc agacgagaat
     2821 gacaaccctc ctcagtttta tccacgggag tatgctgcca gtataagtgc ccagagtcca
     2881 ccaggcacag ctgtgctgag gttgcgtgcc catgaccctg accagggatc ccatgggcga
     2941 ctctcctacc atatcctggc tggcaacagc cccccacttt ttaccttgga tgagcaatca
     3001 gggctgttga cagtagcctg gcccttggcc agacgggcca attctgtggt gcagctggag
     3061 attggggctg aggacggagg tggcctacag gcagaaccca gtgcccgagt ggacatcagc
     3121 attgtgcctg gaacccccac accacccata tttgagcaac tacagtatgt tttttctgtg
     3181 ccagaggatg tggcaccagg caccagtgtg ggcatagtcc aggcacacaa cccaccaggt
     3241 cgcttggcac ctgtgaccct ttccctatca ggtggggatc cccgaggact cttctcccta
     3301 gatgcggtat caggactgtt gcaaacactt cgccctctgg accgggagct actgggacca
     3361 gtgttggagc tggaggtgcg agcaggcagt ggagtgcccc cagctttcgc tgtagctcgg
     3421 gtgcgtgtgc tgctggatga tgtgaatgac aactcccctg cctttcctgc acctgaagac
     3481 acggtattgc taccaccaaa cactgcccca gggactccca tctatacact gcgggctctt
     3541 gaccccgact caggtgttaa cagtcgagtc acctttaccc tgcttgctgg gggtggtgga
     3601 gccttcaccg tggaccccac cacaggccat gtacggctta tgaggcctct ggggccctca
     3661 ggagggccag cccatgagct ggagctggag gcccgggatg ggggctcccc accacgcacc
     3721 agccactttc gactacgggt ggtggtacag gatgtgggaa cccgtgggct ggctccccga
     3781 ttcaacagcc ctacctaccg tgtggacctg ccctcaggca ccactgctgg aactcaggtc
     3841 ctgcaagtgc aggcccaagc accagatggg ggccctatca cctatcacct tgcagcagag
     3901 ggagcaagta gcccctttgg cctggagcca cagagtgggt ggctatgggt gcgggcagca
     3961 ctagaccgtg aggcccagga attgtacata ctgaaggtaa tggcagtgtc tgggtccaaa
     4021 gctgagttgg ggcagcagac aggcacagcc accgtgaggg tcagcatcct caaccagaat
     4081 gaacacagtc cccgcttgtc tgaggatccc accttcctgg ctgtggctga gaaccagccc
     4141 ccagggacca gcgtgggccg agtctttgcc actgaccgag actcaggacc caatggacgt
     4201 ctgacctaca gcctgcaaca gctgtctgaa gacagcaagg ccttccgcat ccacccccag
     4261 actggagaag tgaccacact ccaaaccctg gaccgtgagc agcagagcag ctatcagctc
     4321 ctggtgcagg tgcaggatgg agggagccca ccccgcagca ccacaggcac tgtgcatgtt
     4381 gcagtgcttg acctcaacga caacagcccc acgttcctgc aggcttcagg agctgctggt
     4441 gggggcctcc ctatacaggt accagaccgc gtgcctccag gaacactggt gacgactctg
     4501 caggcgaagg atccagatga gggggagaat gggaccatct tgtacacgct aactggtcct
     4561 ggctcagagc ttttctctct gcaccctcac tcaggggagc tgctcactgc agctcccctg
     4621 atccgagcag agcggcccca ctatgtgctg acactgagtg ctcatgacca aggcagccct
     4681 cctcgaagtg ccagcctcca gctgctggtg caggtgcttc cctcagctcg cttggccgag
     4741 ccgcccccag atctcgcaga gcgggaccca gcggcaccag tgcctgtcgt gctgacggtg
     4801 acagcagctg agggactgcg gcccggctct ctgttgggct cggtggcagc gccagagccc
     4861 gcgggtgtgg gtgcactcac ctacacactg gtgggcggtg ccgatcccga gggcaccttc
     4921 gcgctggatg cggcctcagg gcgcttgtac ctggcgcggc ccctggactt cgaagctggc
     4981 ccgccgtggc gcgcgctcac ggtacgcgct gaggggccgg gaggcgcggg cgcgcggctg
     5041 ctgcgagtgc aggtgcaagt gcaggacgag aatgagcatg cgcccgcctt tgcgcgcgac
     5101 ccgctggcgc tggcgctgcc agagaacccg gagcccggcg cagcgctgta cactttccgc
     5161 gcgtcggacg ccgacggccc cggccccaat agcgacgtgc gctaccgcct gctgcgccag
     5221 gagccgcccg tgccggcgct tcgcctggac gcgcgcaccg gggcgctcag cgctccgcgc
     5281 ggcctggacc gagagaccac tcccgcgctg ctgctgctgg tggaagccac cgaccggccc
     5341 gccaacgcca gccgccgtcg tgcagcgcgc gtttcagcgc gcgtcttcgt cacggatgag
     5401 aatgacaacg cgcctgtctt cgcctcgccg tcacgcgtgc gcctcccaga ggaccagccg
     5461 cctgggcccg cggccctgca cgtggtagcc cgggacccgg atctgggcga ggctgcacgc
     5521 gtgtcctatc ggctggcatc tggcggggac ggccacttcc ggctgcactc aagcactgga
     5581 gcgctgtccg tggtgcggcc gttggaccgc gaacaacgag ctgagcacgt actgacagtg
     5641 gtggcctcag accacggctc cccgccgcgc tcggccacgc aggtcctgac cgtcagtgtc
     5701 gctgacgtca acgacgaggc gcctactttc cagcagcagg agtacagcgt cctcttgcgt
     5761 gagaacaacc ctcctggcac atctctgctc accctgcgag caaccgaccc cgacgtgggg
     5821 gccaacgggc aagtgactta tggaggcgtc tctagcgaaa gcttttctct ggatcctgac
     5881 actggtgttc tcacgactct tcgggccctg gatcgagagg aacaggagga gatcaacctg
     5941 acagtgtatg cccaggacag gggctcacct cctcagttaa cgcatgtcac tgttcgagtg
     6001 gctgtggagg atgagaatga ccatgcacca acctttggga gtgcccatct ctctctggag
     6061 gtgcctgagg gccaggaccc ccagaccctt accatgcttc gggcctctga tccagatgtg
     6121 ggagccaatg ggcagttgca gtaccgcatc ctagatgggg acccatcagg agcctttgtc
     6181 ctagaccttg cttctggaga gtttggcacc atgcggccac tagacagaga agtggagcca
     6241 gctttccagc tgaggataga ggcccgggat ggaggccagc cagctctcag tgccacgctg
     6301 cttttgacag tgacagtgct ggatgccaat gaccatgctc cagcctttcc tgtgcctgcc
     6361 tactcggtgg aggtgccgga ggatgtgcct gcagggaccc tgctgctgca gctacaggct
     6421 catgaccctg atgctggagc taatggccat gtgacctact acctgggcgc cggtacagca
     6481 ggagccttcc tgctggagcc cagctctgga gaactgcgca cagctgcagc cttggacaga
     6541 gaacagtgtc ccagctacac cttttctgtg agtgcagtgg atggtgcagc tgctgggccc
     6601 ctaagcacca cagtgtctgt caccatcatg gtgcgcgatg tcaatgacca tgcacccacc
     6661 ttccccacca gtcctctgcg cctacgtctg ccccgcccag gccccagctt cagtacccca
     6721 accctggctc tggccacact gagagctgaa gatcgtgatg ctggtgccaa tgcttccatt
     6781 ctgtaccggc tggcaggcac accacctcct ggcactactg tggactctta cactggtgaa
     6841 atccgcgtgg cccgctctcc tgtagctcta ggcccccgag atcgtgtcct cttcattgtg
     6901 gccactgatc ttggccgtcc agctcgctct gccactggtg tgatcattgt tggactgcag
     6961 ggggaagctg agcgtggacc ccgctttccc cgggctagca gtgaggctac gattcgtgag
     7021 aatgcgcccc cagggactcc tattgtctcc cccagggccg tccatgcagg aggcacaaat
     7081 ggacccatca cctacagcat tctcagtggg aatgagaaag ggacattctc catccagcct
     7141 agtacaggtg ccatcacagt tcgctcagca gaggggctag acttcgaggt gagtccacgg
     7201 ctgcgactgg tgctgcaggc agagagtgga ggagcctttg ccttcactgt gctgaccctg
     7261 accctgcaag atgccaacga caatgctccc cgtttcctgc ggccccatta tgtggccttc
     7321 cttcctgagt cccggccctt ggaggggccc ctgctgcagg tggaggcgga tgacctggat
     7381 caaggctctg gaggacagat ttcctacagt ctggctgcat cccagccggc acgtggattg
     7441 ttccacgtag acccaaccac aggcactatc actaccacag ccatcctgga ccgtgagatc
     7501 tgggctgaaa cacggttggt gctgatggcc acagacagag ggagcccagc cctggtgggc
     7561 tcagctacct tgacggtgat ggtcatcgac accaatgaca atcgccccac catcccccaa
     7621 ccctgggagc tccgagtgtc agaagatgcg ttattgggct cagagattgc acaggtaaca
     7681 gggaatgatg tggactcagg acccgtgctg tggtatgtgc taagcccatc tgggccccag
     7741 gatcccttca gtgttggccg ctatggaggc cgtgtctccc tcacggggcc cctggacttt
     7801 gagcagtgtg accgctacca gctgcagctg ctggcacatg atgggcctca tgagggccgt
     7861 gccaacctca cagtgcttgt ggaggatgtc aatgacaatg cacctgcctt ctcacagagc
     7921 ctctaccagg taatgctgct tgagcacaca cccccaggca gtgccattct ctccgtctct
     7981 gccactgatc gggactcagg tgccaacggt cacatttcct accacctggc ttcccctgcc
     8041 gatggcttca gtgttgaccc caacaatggg accctgttca caatagtggg aacagtggcc
     8101 ttgggccatg acgggtcagg agcagtggat gtggtgctgg aagcacgaga ccacggggct
     8161 ccaggccggg cagcacgagc cacagtgcac gtgcagctgc aggaccagaa cgaccacgcc
     8221 ccgagcttca cattgtcaca ctaccgtgtg gctgtgactg aagacctgcc ccctggctcc
     8281 actctgctca ccctggaggc tacagatgct gatggaagcc gcagccatgc cgctgtggac
     8341 tacagcatca tcagtggcaa ctggggccga gtcttccagc tggaacccag gctggctgag
     8401 gctggggaga gtgctggacc aggcccccgg gcactgggct gcctggtgtt gcttgaacct
     8461 ctagactttg aaagcctgac acagtacaat ctaacagtgg ctgcagctga ccgtgggcag
     8521 ccaccccaaa gctcagtcgt gccagtcact gtcactgtac tagatgtcaa tgacaaccca
     8581 cctgtcttta cccgagcatc ctaccgtgtg acagtacctg aggacacacc tgttggagct
     8641 gagctgctgc atgtagaggc ctctgacgct gaccctggcc ctcatggcct cgtgcgtttc
     8701 actgtcagct caggcgaccc atcagggctc tttgagctgg atgagagctc aggcaccttg
     8761 cgactggccc atgccctgga ctgtgagacc caggctcgac atcagcttgt agtacaggct
     8821 gctgaccctg ctggtgcaca ctttgctttg gcaccagtga caattgaggt ccaggatgtg
     8881 aatgatcatg gcccagcctt cccactgaac ttactcagca ccagcgtggc cgagaatcag
     8941 cctccaggca ctctcgtgac cactctgcat gcaatcgacg gggatgctgg ggcttttggg
     9001 aggctccgtt acagcctgtt ggaggctggg ccaggacctg agggccgtga ggcatttgca
     9061 ctgaacagct caacagggga gttgcgtgcg cgagtgccct ttgactatga gcacacagaa
     9121 agcttccggc tgctggtggg tgctgctgat gctgggaatc tctcagcctc tgtcaccgtg
     9181 tcggtgctag tgactggaga ggatgagtat gaccctgtat ttctggcacc agctttccac
     9241 ttccaagtgc ccgaaggtgc ccggcgtggc cacagcttgg gtcacgtgca ggccacagat
     9301 gaggatgggg gtgccgatgg cctggttctg tattcccttg ccacctcttc cccctatttt
     9361 ggtattaacc agactacagg agccctgtac ctgcgggtgg acagtcgggc accaggcagc
     9421 ggaacagcca cctctggggg tgggggccgg acccggcggg aagcaccacg ggagctgagg
     9481 ctggaggtga tagcacgggg gcctctgcct ggttcccgga gtgccacagt gcctgtgacc
     9541 gtggatatca cccacaccgc actgggcctg gcacctgacc tcaacctgct attagtaggg
     9601 gccgtggcag cctccttggg agttgtggtg gtgcttgcac tggcagccct ggtcctagga
     9661 cttgttcggg cccgtagccg caaggctgag gcagcccctg gcccaatgtc acaggcagca
     9721 cccctagcca gtgactcact gcagaaactg ggccgggagc cacctagtcc accaccctct
     9781 gagcacctct atcaccagac tcttcccagc tatggtgggc caggagctgg aggaccctac
     9841 ccccgtggtg gctccttgga cccttcacat tcaagtggcc gaggatcagc agaggctgca
     9901 gaggatgatg agatccgcat gatcaatgag ttcccccgtg tggccagtgt ggcctcctct
     9961 ctggctgccc gtggccctga ctcaggcatc cagcaggatg cagatggtct gagtgacaca
    10021 tcctgcgaac cacctgcccc tgacacctgg tataagggcc gaaaggcagg gctgctgctg
    10081 ccaggtgcag gagccactct ctacagagag gaggggcccc cagccactgc cacagccttc
    10141 ctggggggct gtggcctgag ccctgcaccc actggggact atggcttccc agcagatggc
    10201 aagccatgtg tggcaggtgc gctgacagcc attgtggccg gcgaggagga gctccgtggc
    10261 agctataact gggactacct gctgagctgg tgccctcagt tccaaccact ggccagtgtc
    10321 ttcacagaga tcgctcggct caaggatgaa gctcggccat gtcccccagc tccccgtatc
    10381 gacccaccac ccctcatcac tgccgtggcc cacccaggag ccaagtctgt gccccccaag
    10441 ccagcaaaca cagctgcagc ccgggccatc ttcccaccag cttctcaccg ctcccccatc
    10501 agccatgaag gctccctgtc ctcagctgcc atgtccccca gcttctcacc ctctctgtct
    10561 cctctggctg ctcgctcacc cgttgtctca ccatttgggg tggcccaggg tccctcagcc
    10621 tcagcactca gcgcagagtc tggcctggag ccacctgatg acacggagct gcacatcacc
    10681 ggtggctctg gcatggtgag caagggcgag gaggataaca tggccatcat caaggagttc
    10741 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
    10801 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
    10861 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
    10921 tacgtgaagc accccgccga cattcccgac tacttgaagc tgtccttccc cgagggcttc
    10981 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
    11041 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
    11101 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
    11161 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
    11221 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
    11281 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
    11341 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
    11401 tcttgactcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
    11461 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
    11521 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
    11581 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
    11641 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
    11701 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
    11761 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
    11821 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
    11881 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
    11941 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
    12001 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
    12061 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
    12121 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
    12181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
    12241 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
    12301 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
    12361 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
    12421 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
    12481 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtga
    12541 tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
    12601 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
    12661 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
    12721 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
    12781 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
    12841 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
    12901 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
    12961 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
    13021 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
    13081 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
    13141 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
    13201 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
    13261 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
    13321 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
    13381 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
    13441 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
    13501 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
    13561 aatagcacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
    13621 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
    13681 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
    13741 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
    13801 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
    13861 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
    13921 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
    13981 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
    14041 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
    14101 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
    14161 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
    14221 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
    14281 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
    14341 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
    14401 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
    14461 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
    14521 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
    14581 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
    14641 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
    14701 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
    14761 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
    14821 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
    14881 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
    14941 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
    15001 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
    15061 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
    15121 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
    15181 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
    15241 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
    15301 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
    15361 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
    15421 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
    15481 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
    15541 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
    15601 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
    15661 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
    15721 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
    15781 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
    15841 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
    15901 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
    15961 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
    16021 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
    16081 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
    16141 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
    16201 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//

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