质粒名称: | pCDH-EFIa-Id1-PGK-puro |
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出品公司: | Addgene |
目录编号: | 107735 |
存储实验室: | Alexandre Colas |
载体骨架基本信息 |
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载体名称: | pCDH-EF1mcs-PGKpuro |
载体出品公司: | System Biosciences |
空载体大小: | 8650 bp |
完整质粒大小: | 9589 bp |
载体类型: | Lentiviral |
筛选标记: | Puromycin |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | Id1 |
物种来源: | M. musculus (mouse) |
插入大小: | 939 bp |
基因突变: | Used pCDH-CMV-MCS-copGFP stock plasmid CD511B-1 from System Biosciences. Modified by replacing CMV promoter with EF1 and replacing copGFP with PGKpuro |
GenBank编号 | NM_010495 NM_010495 |
启动子: | EF1a |
克隆方法: | 限制性内切酶 |
5’克隆位点: | XbaI (unknown if destroyed) |
3’克隆位点: | EcoRI (unknown if destroyed) |
5’测序引物 | ATGAAGGTCGCCAGTGGCAGTGCC |
3’测序引物 | TCAGCGACACAAGATGCGATCGTC |
文献引用方法 |
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材料和方法部分: | pCDH-EFIa-Id1-PGK-puro was a gift from Alexandre Colas (Addgene plasmid # 107735 ; http://n2t.net/addgene:107735 ; RRID:Addgene_107735) |
参考文献部分: | Id genes are essential for early heart formation. Cunningham TJ, Yu MS, McKeithan WL, Spiering S, Carrette F, Huang CT, Bushway PJ, Tierney M, Albini S, Giacca M, Mano M, Puri PL, Sacco A, Ruiz-Lozano P, Riou JF, Umbhauer M, Duester G, Mercola M, Colas AR. Genes Dev. 2017 Aug 9. pii: gad.300400.117. doi: 10.1101/gad.300400.117. 10.1101/gad.300400.117 PubMed 28794185 |
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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 pCDH-EFIa-Id1-PGK-puro 9171 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses Id1 in mammalian cells. ACCESSION . VERSION . KEYWORDS pCDH-EFIa-Id1-PGK-puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9171) AUTHORS Cunningham TJ, Yu MS, McKeithan WL, Spiering S, Carrette F, Huang CT, Bushway PJ, Tierney M, Albini S, Giacca M, Mano M, Puri PL, Sacco A, Ruiz-Lozano P, Riou JF, Umbhauer M, Duester G, Mercola M, Colas AR TITLE Id genes are essential for early heart formation. JOURNAL Genes Dev. 2017 Aug 9. pii: gad.300400.117. doi: 10.1101/gad.300400.117. PUBMED 28794185 REFERENCE 2 (bases 1 to 9171) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..9171 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 2..24 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 16..33 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 17..33 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 48..274 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 275..455 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 502..627 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1120..1353 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1538..1582 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 1849..1965 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2014..2225 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 2238..2506 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" promoter 2577..2788 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 2801..3069 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" promoter 3140..3351 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 3364..3632 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" promoter 4154..4664 /label=hPGK promoter /note="human phosphoglycerate kinase 1 promoter" CDS 4686..5285 /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(4686..4705) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5182..5202 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 5292..5880 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5345..5365) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5763..5774) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5954..6187 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 6259..6380 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6296..6315) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6350..6369 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 6420..6555 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6482..6501 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(6588..6604) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6588..6604) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6601..6623) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6612..6628 /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(6636..6666) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6681..6702 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6819..6836) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6990..7578) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7070..7089) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7749..8609) /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 8372..8391 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8610..8714) /gene="bla" /label=AmpR promoter primer_bind 8782..8800 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(8838..8860) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 8960..8979 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgacgcgtgt agtcttatgc 61 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga 121 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg 181 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga 241 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg 301 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc 361 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct 421 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa 481 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg 541 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag 601 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg 661 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg 721 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc 781 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga 841 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac 901 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag 961 caagcggcca ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga 1021 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa 1081 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt 1141 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag 1201 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca 1261 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc 1321 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact 1381 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat 1441 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat 1501 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga 1561 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat 1621 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact 1681 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc 1741 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag 1801 agacagatcc attcgattag tgaacggatc tcgacggtat cggttaactt ttaaaagaaa 1861 aggggggatt ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat 1921 acaaactaaa gaattacaaa aacaaattac aaaattcaaa attttatcga tactagtccg 1981 caaggatctg cgatcgctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc 2041 ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc ctagagaagg tggcgcgggg 2101 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac 2161 cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa 2221 cacagctgaa gcttcgaggg gctcgcatct ctccttcacg cgcccgccgc cctacctgag 2281 gccgccatcc acgccggttg agtcgcgttc tgccgcctcc cgcctgtggt gcctcctgaa 2341 ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag accgggcctt tgtccggcgc 2401 tcccttggag cctacctaga ctcagccggc tctccacgct ttgcctgacc ctgcttgctc 2461 aactctacgt ctttgtttcg ttttctgttc tgcgccgtta cagatccaag ctgtgaccgg 2521 cgcctacgct agactctagt ccgcaaggat ctgcgatcgc tccggtgccc gtcagtgggc 2581 agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg 2641 tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct 2701 ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt 2761 tcgcaacggg tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc 2821 acgcgcccgc cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc 2881 tcccgcctgt ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc 2941 gagaccgggc ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac 3001 gctttgcctg accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg 3061 ttacagatcc aagctgtgac cggcgcctac gctagactct agtccgcaag gatctgcgat 3121 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg 3181 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt 3241 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca 3301 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt 3361 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc 3421 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc 3481 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta 3541 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt 3601 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac 3661 tctagagcca ccatgaaggt cgccagtggc agtgccgcag ccgctgcagg ccctagctgt 3721 tcgctgaagg cgggcaggac agcgggcgag gtggtacttg gtctgtcgga gcaaagcgtg 3781 gccatctcgc gctgcgctgg gacgcgcctg cccgccttgc tggacgagca gcaggtgaac 3841 gtcctgctct acgacatgaa cggctgctac tcacgcctca aggagctggt gcccaccctg 3901 ccccagaacc gcaaagtgag caaggtggag atcctgcagc atgtaatcga ctacatcagg 3961 gacctgcagc tggagctgaa ctcggagtct gaagtcggga ccaccggagg ccggggactg 4021 cctgtccgcg ccccgctcag caccctgaac ggcgagatca gtgccttggc ggccgaggcg 4081 gcatgtgttc cagccgacga tcgcatcttg tgtcgctgag aattcgaatt taaatcggat 4141 ccgcggccgc cacggggttg gggttgcgcc ttttccaagg cagccctggg tttgcgcagg 4201 gacgcggctg ctctgggcgt ggttccggga aacgcagcgg cgccgaccct gggtctcgca 4261 cattcttcac gtccgttcgc agcgtcaccc ggatcttcgc cgctaccctt gtgggccccc 4321 cggcgacgct tcctgctccg cccctaagtc gggaaggttc cttgcggttc gcggcgtgcc 4381 ggacgtgaca aacggaagcc gcacgtctca ctagtaccct cgcagacgga cagcgccagg 4441 gagcaatggc agcgcgccga ccgcgatggg ctgtggccaa tagcggctgc tcagcagggc 4501 gcgccgagag cagcggccgg gaaggggcgg tgcgggaggc ggggtgtggg gcggtagtgt 4561 gggccctgtt cctgcccgcg cggtgttccg cactctgcaa gcctccggag cgcacgtcgg 4621 cagtcggctc cctcgttgac cgaatcaccg acctctctcc ccaggggaat ccaccggacg 4681 ccaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtcccccggg 4741 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgttgacc 4801 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc 4861 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc 4921 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga 4981 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca 5041 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc 5101 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct 5161 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca 5221 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg 5281 cctgagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta 5341 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta 5401 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt 5461 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg 5521 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt 5581 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag 5641 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc 5701 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc 5761 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc 5821 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc 5881 ctggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa 5941 gaaaaggggg gactggaagg gctaattcac tcccaacgaa gataagatct gctttttgct 6001 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg 6061 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt 6121 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc 6181 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga 6241 atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa ataaagcaat 6301 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 6361 aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta actccgccca 6421 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 6481 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 6541 gcttttttgg aggcctagac ttttgcagag accaaattcg taatcatgtc atagctgttt 6601 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag 6661 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg 6721 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg 6781 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 6841 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 6901 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 6961 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 7021 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 7081 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 7141 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 7201 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 7261 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 7321 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 7381 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 7441 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 7501 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 7561 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 7621 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 7681 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 7741 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 7801 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 7861 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 7921 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 7981 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 8041 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 8101 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 8161 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 8221 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 8281 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 8341 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 8401 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 8461 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 8521 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 8581 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 8641 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 8701 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt 8761 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc 8821 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg 8881 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt 8941 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg 9001 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc 9061 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg 9121 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt t //