质粒名称: | pLenti X2 Puro DEST (w16-1) |
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出品公司: | 爱迪基因 |
目录编号: | 17296 |
存储实验室: | Eric Campeau, Paul Kaufman |
载体骨架基本信息 |
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载体名称: | p156RRL-sinPPT-CMV-GFP-PRE/Nhe I |
载体抗性: | 氯霉素和氨苄霉素 |
空载体大小: | 9070 bp |
载体类型: | Lentiviral, RNAi ; Destination vector |
筛选标记: | 嘌呤霉素 |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | None |
克隆方法: | 限制性内切酶 |
5' 测序引物 | n/a |
文献引用方法 |
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材料和方法部分: | pLenti X2 Puro DEST (w16-1) was a gift from Eric Campeau & Paul Kaufman (爱迪基因 plasmid # 17296) |
参考文献部分: | A versatile viral system for expression and depletion of proteins in mammalian cells. Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD.. PLoS One. 2009 Aug 6;4(8):e6529. 10.1371/journal.pone.0006529 PubMed 19657394 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pLenti X2 Puro DEST (w16-1) 9070 bp ds-DNA circular SYN 10-JAN-2018 DEFINITION 3rd gen lentiviral Gateway destination vector, RNAi, 3'LTR insertion, Puro. ACCESSION . VERSION . KEYWORDS pLenti X2 Puro DEST (w16-1) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9070) AUTHORS Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, Campisi J, Yaswen P, Cooper PK, Kaufman PD. TITLE A versatile viral system for expression and depletion of proteins in mammalian cells. JOURNAL PLoS One. 2009 Aug 6;4(8):e6529. PUBMED 19657394 COMMENT This file is created by Vector NTI http://www.biofeng.com/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|biofeng.com| COMMENT VNTNAME|pLenti X2 Puro DEST (w16-1)| FEATURES Location/Qualifiers source 1..9070 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 27..131 /gene="bla" /label=AmpR promoter CDS 132..992 /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(350..369) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1163..1751 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1652..1671 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1905..1922 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2039..2060 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2075..2105 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2113..2129 /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 2118..2140 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2137..2153 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2137..2153 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 2172..2192 /label=T3 /note="T3 promoter, forward primer" promoter 2174..2192 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" LTR 2448..2628 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 2672..2797 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 3290..3523 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 4012..4129 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" misc_feature 4183..4771 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4236..4256) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4654..4665) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" promoter 4785..5284 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(4820..4843) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 5217..5236 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 5305..5904 /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(5305..5324) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5801..5821 /label=Puro-F /note="Puromycin resistance gene, forward primer" protein_bind 6085..6209 /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS complement(6250..6555) /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 complement(6352..6371) /label=ccdB-fwd /note="ccdB gene, forward primer" CDS complement(6897..7556) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 7471..7490 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(7610..7640) /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind complement(7665..7789) /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" LTR 7864..8044 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" polyA_signal 8116..8237 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8153..8172) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8207..8226 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 8256..8391 /label=SV40 ori /note="SV40 origin of replication" primer_bind 8318..8337 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(8423..8442) /label=T7 /note="T7 promoter, forward primer" promoter complement(8424..8442) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(8452..8469) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(8452..8468) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8461..8483) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 8613..9068 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8700..8719) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8910..8931 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac 61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg 1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt 1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta 2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta 2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt 2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat 2221 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc 2281 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg 2341 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc 2401 gcattgcaga gatattgtat ttaagtgcct agctcgatac aataaacggg tctctctggt 2461 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc 2521 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta 2581 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa 2641 cagggacctg aaagcgaaag ggaaaccaga gctctctcga cgcaggactc ggcttgctga 2701 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag 2761 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag 2821 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca 2881 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac 2941 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga 3001 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga 3061 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac 3121 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt 3181 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca 3241 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt 3301 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcctcaatg acgctgacgg 3361 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta 3421 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa 3481 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct 3541 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc 3601 tggaacagat tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac 3661 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga 3721 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct 3781 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt 3841 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac 3901 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga 3961 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggttaa cttttaaaag 4021 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga 4081 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta ttccagtgtg 4141 gtggaattct gcagatatcc agcacagtgg cggccgctcg acaatcaacc tctggattac 4201 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 4261 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 4321 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 4381 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 4441 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 4501 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 4561 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg 4621 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 4681 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 4741 agtcggatct ccctttgggc cgcctccccg cctggaattc taccgggtag gggaggcgct 4801 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac 4861 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cggctccgtt 4921 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc 4981 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt 5041 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata 5101 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg 5161 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt 5221 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct 5281 ttcgacctgc agcccaagct taccatgacc gagtacaagc ccacggtgcg cctcgccacc 5341 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc 5401 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc 5461 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg 5521 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc 5581 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc 5641 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc 5701 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag 5761 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag 5821 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc 5881 atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc gaccgaaagg 5941 agcgcacgac cccatgcatc tcgagggccc ggtaccttta agaccaatga cttacaaggc 6001 agctgtagat cttagccact ttttaaaaga aaagggggga ctggaagggc tagactcgag 6061 cggccgccac tgtgctggat atcaaccact ttgtacaaga aagctgaacg agaaacgtaa 6121 aatgatataa atatcaatat attaaattag attttgcata aaaaacagac tacataatac 6181 tgtaaaacac aacatatcca gtcactatgg tcgacctgca gactggctgt gtataacgga 6241 gcctgacatt tatattcccc agaacatcag gttaatggcg tttttgatgt cattttcgcg 6301 gtggctgaga tcagccactt cttccccgat aacggagacc ggcacactgg ccatatcggt 6361 ggtcatcatg cgccagcttt catccccgat atgcaccacc gggtaaagtt cacgggagac 6421 tttatctgac agcagacgtg cactggccag ggggatcacc atccgtcgcc cgggcgtgtc 6481 aataatatca ctctgtacat ccacaaacag acgataacgg ctctctcttt tataggtgta 6541 aaccttaaac tgcatttcac cagtccctgt tctcgtcagc aaaagagccg ttcatttcaa 6601 taaaccgggc gacctcagcc atcccttcct gattttccgc tttccagcgt tcggcacgca 6661 gacgacgggc ttcattctgc atggttgtgc ttaccagacc ggagatattg acatcatata 6721 tgccttgagc aactgatagc tgtcgctgtc aactgtcact gtaatacgct gcttcatagc 6781 acacctcttt ttgacatact tcgggtatac atatcagtat atattcttat accgcaaaaa 6841 tcagcgcgca aatacgcata ctgttatctg gcttttagta agccggatcc agatctttac 6901 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg 6961 aagccatcac agacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct 7021 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg 7081 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca 7141 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat 7201 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca 7261 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg 7321 tctttcattg ccatacggaa ttccggatga gcattcatca ggcgggcaag aatgtgaata 7381 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc 7441 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct 7501 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta 7561 gcttccttag ctcctgaaaa tctcgccgga tcctaactca aaatccacac attatacgag 7621 ccggaagcat aaagtgtaaa gcctggggtg cctaatgcgg ccgccatagt gactggatat 7681 gttgtgtttt acagtattat gtagtctgtt ttttatgcaa aatctaattt aatatattga 7741 tatttatatc attttacgtt tctcgttcag cttttttgta caaacttgtt gatatctgca 7801 gaattccacc acactggact agctcactcc caacgaagac aagatctgct ttttgcttgt 7861 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggctg cctagggaac 7921 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg 7981 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct 8041 agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa gaaatgaata 8101 tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata aagcaatagc 8161 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa 8221 ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact ccgcccagtt 8281 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 8341 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 8401 gcgtcgagac gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt 8461 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc 8521 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca 8581 acagttgcgc agcctgaatg gcgaatggcg cgacgcgccc tgtagcggcg cattaagcgc 8641 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc 8701 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct 8761 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa 8821 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc 8881 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact 8941 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg 9001 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt 9061 tacaatttcc //