pAAVS1-P-CAG-DEST

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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: pAAVS1-P-CAG-DEST
出品公司: Addgene
目录编号: 80490
存储实验室: Knut Woltjen

载体骨架基本信息
载体名称: pBluescript
载体出品公司: --
空载体大小: 3000 bp
完整质粒大小: --
载体修饰: AAVS1的同源臂;无启动子SA-T2A-puro选择盒;CAG启动子;网关目的磁带
载体类型: 哺乳动物表达;供体载体(人类)
筛选标记: 嘌呤霉素
载体抗性: 氯霉素和氨苄青霉素
生长温度: 37 ℃
宿主菌: ccdB Survival
拷贝数: 高拷贝

插入基因信息
插入基因名称: --
物种来源: --
插入大小: --
基因突变: --
GenBank编号 --
启动子: --
标签/融合蛋白: --
克隆方法: Gateway克隆
5’克隆位点: --
3’克隆位点: --
5’测序引物 --
3’测序引物 --

文献引用方法
材料和方法部分: pAAVS1-P-CAG-DEST was a gift from Knut Woltjen (Addgene plasmid # 80490 ; http://n2t.net/addgene:80490 ; RRID:Addgene_80490)
参考文献部分: Engineering the AAVS1 locus for consistent and scalable transgene expression in human iPSCs and their differentiated derivatives. Oceguera-Yanez F, Kim SI, Matsumoto T, Tan GW, Xiang L, Hatani T, Kondo T, Ikeya M, Yoshida Y, Inoue H, Woltjen K. Methods. 2015 Dec 18. pii: S1046-2023(15)30181-X. doi: 10.1016/j.ymeth.2015.12.012. 10.1016/j.ymeth.2015.12.012 PubMed 26707206

载体图谱质粒图谱和多克隆位点信息

pAAVS1-P-CAG-DEST质粒图谱

载体简介

载体序列

LOCUS       pAAVS1-P-CAG-DEST              9502 bp ds-DNA     circular SYN 09-SEP-2021
DEFINITION  Gateway Destination donor vector for AAVS1 targeting (puromycin 
            selection).
ACCESSION   .
VERSION     .
KEYWORDS    pAAVS1-P-CAG-DEST
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9502)
  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 9502)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 9, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9502
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     intron          105..1121
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1045..1067
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1129..1148
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1215..1339
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1364..1394
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1448..2107
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1514..1533)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2449..2754
                     /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="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     2633..2652
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2795..2919)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3015..3034)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3080..3135
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3134..3153)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     misc_feature    3691..4220
                     /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(4366..4385)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4367..4385)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4392..4409)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4392..4408)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4401..4423)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4550..5005
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4637..4656)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4847..4868
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5031..5135
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5136..5996
                     /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     complement(5354..5373)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6167..6755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6656..6675
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6909..6926
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7043..7064
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7079..7109
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7117..7133
                     /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     7122..7144
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7141..7157
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7141..7157
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7178..7196
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    7209..8012
                     /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    8019..8044
                     /label=SA
                     /note="splice acceptor site"
     CDS             8068..8121
                     /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             8131..8730
                     /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(8131..8150)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8627..8647
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     complement(8762..8779)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8768..8992
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        9011..9390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
       61 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
      121 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
      181 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
      241 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
      301 gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
      361 ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
      421 ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
      481 gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
      541 cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg
      601 ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg
      661 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
      721 gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg
      781 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
      841 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
      901 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
      961 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     1021 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag
     1081 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg
     1141 ttattgtgct gtctcatcat tttggcaaag aattcgcggc cgcggtaccg gcgcgccgga
     1201 tcctgcaggt tatcacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa
     1261 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
     1321 aacatatcca gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc
     1381 cggctcgtat aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga
     1441 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     1501 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     1561 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     1621 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
     1681 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     1741 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     1801 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     1861 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     1921 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     1981 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
     2041 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     2101 ggcgtaaacg cgtggatccg gcttactaaa agccagataa cagtatgcgt atttgcgcgc
     2161 tgatttttgc ggtataagaa tatatactga tatgtatacc cgaagtatgt caaaaagagg
     2221 tatgctatga agcagcgtat tacagtgaca gttgacagcg acagctatca gttgctcaag
     2281 gcatatatga tgtcaatatc tccggtctgg taagcacaac catgcagaat gaagcccgtc
     2341 gtctgcgtgc cgaacgctgg aaagcggaaa atcaggaagg gatggctgag gtcgcccggt
     2401 ttattgaaat gaacggctct tttgctgacg agaacagggg ctggtgaaat gcagtttaag
     2461 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt
     2521 attgacacgc ccgggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat
     2581 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg
     2641 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc
     2701 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca
     2761 ggctccctta tacacagcca gtctgcaggt cgaccatagt gactggatat gttgtgtttt
     2821 acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc
     2881 attttacgtt tctcgttcag ctttcttgta caaagtggtg ataactctag agaattcact
     2941 cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac
     3001 aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     3061 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     3121 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     3181 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     3241 tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat
     3301 agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct
     3361 ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga
     3421 ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag
     3481 cttggatccc tcgagttaat taaatctaga agtcgacagt actaagcttt gacagaaaag
     3541 ccccatcctt aggcctcctc cttcctagtc tcctgatatt gggtctaacc cccacctcct
     3601 gttaggcaga ttccttatct ggtgacacac ccccatttcc tggagccatc tctctccttg
     3661 ccagaacctc taaggtttgc ttacgatgga gccagagagg atcctgggag ggagagcttg
     3721 gcagggggtg ggagggaagg gggggatgcg tgacctgccc ggttctcagt ggccaccctg
     3781 cgctaccctc tcccagaacc tgagctgctc tgacgcggct gtctggtgcg tttcactgat
     3841 cctggtgctg cagcttcctt acacttccca agaggagaag cagtttggaa aaacaaaatc
     3901 agaataagtt ggtcctgagt tctaactttg gctcttcacc tttctagtcc ccaatttata
     3961 ttgttcctcc gtgcgtcagt tttacctgtg agataaggcc agtagccagc cccgtcctgg
     4021 cagggctgtg gtgaggaggg gggtgtccgt gtggaaaact ccctttgtga gaatggtgcg
     4081 tcctaggtgt tcaccaggtc gtggccgcct ctactccctt tctctttctc catccttctt
     4141 tccttaaaga gtccccagtg ctatctggga catattcctc cgcccagagc agggtcccgc
     4201 ttccctaagg ccctgctctg ggcttctggg tttgagtcct tggcaagccc aggagaggcg
     4261 ctcaggcttc cctgtccccc ttcctcgtcc accatctcat gcccctggct ctcctgcccc
     4321 ttccctacag gggttcctgg ctctgctcta gcgatcgcca attcgcccta tagtgagtcg
     4381 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     4441 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     4501 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     4561 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4621 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4681 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4741 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4801 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4861 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4921 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4981 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     5041 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     5101 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     5161 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     5221 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     5281 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     5341 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     5401 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     5461 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     5521 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     5581 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     5641 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     5701 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     5761 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     5821 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     5881 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     5941 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     6001 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6061 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6121 cgttccactg agcgtcaaac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6181 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6241 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6301 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6361 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6421 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     6481 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     6541 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     6601 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     6661 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     6721 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     6781 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     6841 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     6901 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     6961 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     7021 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     7081 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     7141 caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac taaagggaac
     7201 aaaagctgtg ctttctctga ccagcattct ctcccctggg cctgtgccgc tttctgtctg
     7261 cagcttgtgg cctgggtcac ctctacggct ggcccagatc cttccctgcc gcctccttca
     7321 ggttccgtct tcctccactc cctcttcccc ttgctctctg ctgtgttgct gcccaaggat
     7381 gctctttccg gagcacttcc ttctcggcgc tgcaccacgt gatgtcctct gagcggatcc
     7441 tccccgtgtc tgggtcctct ccgggcatct ctcctccctc acccaacccc atgccgtctt
     7501 cactcgctgg gttccctttt ccttctcctt ctggggcctg tgccatctct cgtttcttag
     7561 gatggccttc tccgacggat gtctcccttg cgtcccgcct ccccttcttg taggcctgca
     7621 tcatcaccgt ttttctggac aaccccaaag taccccgtct ccctggcttt agccacctct
     7681 ccatcctctt gctttctttg cctggacacc ccgttctcct gtggattcgg gtcacctctc
     7741 actcctttca tttgggcagc tcccctaccc cccttacctc tctagtctgt gctagctctt
     7801 ccagccccct gtcatggcat cttccagggg tccgagagct cagctagtct tcttcctcca
     7861 acccgggccc ctatgtccac ttcaggacag catgtttgct gcctccaggg atcctgtgtc
     7921 cccgagctgg gaccacctta tattcccagg gccggttaat gtggctctgg ttctgggtac
     7981 ttttatctgt cccctccacc ccacagtggg gcaagcttct gacctcttct cttcctccca
     8041 cagggcctcg agagatctgg cagcggagag ggcagaggaa gtcttctaac atgcggtgac
     8101 gtggaggaga atcccggccc taggctcgag atgaccgagt acaagcccac ggtgcgcctc
     8161 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     8221 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     8281 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     8341 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     8401 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     8461 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     8521 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     8581 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     8641 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     8701 tggtgcatga cccgcaagcc cggtgcctga tctagagggc ccgtttaaac ccgctgatca
     8761 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     8821 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     8881 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     8941 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggactagcgg
     9001 tggcggcctc gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     9061 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     9121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     9181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     9241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     9301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     9361 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     9421 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     9481 tgtgcagcga tgggggcggg gg
//


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