质粒名称: | pAY5 (mAID-EGFP-NLS piggyBac) |
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出品公司: | Addgene |
目录编号: | 140532 |
存储实验室: | Masato Kanemaki |
载体骨架基本信息 |
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载体名称: | pPB-LR5 |
载体出品公司: | -- |
空载体大小: | -- |
完整质粒大小: | 8447 bp |
载体修饰: | -- |
载体类型: | 哺乳动物表达 |
筛选标记: | 潮霉素 |
载体抗性: | 氨苄青霉素,100μg/mL |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | mAID-EGFP-NLS |
物种来源: | H. sapiens (human) |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子: | -- |
标签/融合蛋白: | -- |
文献引用方法 |
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材料和方法部分: | pAY5 (mAID-EGFP-NLS piggyBac) was a gift from Masato Kanemaki (Addgene plasmid # 140532 ; http://n2t.net/addgene:140532 ; RRID:Addgene_140532) |
参考文献部分: | The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice. Yesbolatova A, Saito Y, Kitamoto N, Makino-Itou H, Ajima R, Nakano R, Nakaoka H, Fukui K, Gamo K, Tominari Y, Takeuchi H, Saga Y, Hayashi KI, Kanemaki MT. Nat Commun. 2020 Nov 11;11(1):5701. doi: 10.1038/s41467-020-19532-z. 10.1038/s41467-020-19532-z PubMed 33177522 |
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载体基本信息
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载体序列
LOCUS pAY5 (mAID-EGFP-NLS piggyBac) 8447 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION mAID-EGFP-NLS piggyBac. ACCESSION . VERSION . KEYWORDS pAY5 (mAID-EGFP-NLS piggyBac) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8447) AUTHORS Yesbolatova A, Saito Y, Kitamoto N, Makino-Itou H, Ajima R, Nakano R, Nakaoka H, Fukui K, Gamo K, Tominari Y, Takeuchi H, Saga Y, Hayashi KI, Kanemaki MT. TITLE The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice. JOURNAL Nat Commun (2020) 11(1):5701 REFERENCE 2 (bases 1 to 8447) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..8447 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(3..458) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(140..161) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 352..371 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 585..607 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 599..616 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 600..616 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 626..645 /label=T7 /note="T7 promoter, forward primer" promoter 626..644 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" repeat_region 674..708 /label=piggyBac left (5') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" promoter 1154..2335 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 1384..2326 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 2283..2303 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 2400..2603 /codon_start=1 /product="minimal degron from the auxin-responsive IAA17 protein of Arabidopsis thaliana (Kubota et al., 2013)" /label=mini-AID /note="acts as an auxin-inducible degron (AID) in the presence of TIR1" /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA AAFVKVSMDGAPYLRKIDLRMYK" CDS 2610..3326 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 2610..3326 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2655..2676) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2916..2935) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3263..3284 /label=EGFP-C /note="EGFP, forward primer" CDS 3363..3383 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 3519..3640 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3556..3575) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3610..3629 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind complement(3671..3704) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter 3765..4264 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(3800..3823) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 4197..4216 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 4325..5344 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="KPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYV LRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPET ELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTV MDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGD SQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNF DDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE" protein_bind complement(5907..5940) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." repeat_region 6146..6208 /label=piggyBac right (3') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" primer_bind complement(6259..6279) /label=T3 /note="T3 promoter, forward primer" promoter complement(6259..6277) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(6298..6314) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6298..6314) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6311..6333) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6322..6338 /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(6346..6376) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6391..6412 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6529..6546) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6700..7288) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6780..6799) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7459..8319) /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 8082..8101 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8320..8424) /gene="bla" /label=AmpR promoter ORIGIN 1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg 481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg 541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg 601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggggcgcgc 661 cattctagat taaccctaga aagatagtct gcgtaaaatt gacgcatgca ttcttgaaat 721 attgctctct ctttctaaat agcgcgaatc cgtcgctgtg catttaggac atctcagtcg 781 ccgcttggag ctcccgtgag gcgtgcttgt caatgcggta agtgtcactg attttgaact 841 ataacgaccg cgtgagtcaa aatgacgcat gattatcttt tacgtgactt ttaagattta 901 actcatacga taattatatt gttatttcat gttctactta cgtgataact tattatatat 961 atattttctt gttatagata tcaactagaa tgctagtaat gagtaattca tacaaaagga 1021 ctcgcccctg ccttggggaa tcccagggac cgtcgttaaa ctcccactaa cgtagaaccc 1081 agagatcgct gcgttcccgc cccctcaccc gcccgctctc gtcatcactg aggtggagaa 1141 gagcatgcgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 1201 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta 1261 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 1321 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 1381 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc 1441 gtgccttgaa ttacttccac gcccctggct gcagtacgtg attcttgatc ccgagcttcg 1501 ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc 1561 ttgagttgag gcctggcttg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg 1621 cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc 1681 gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat 1741 ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc 1801 gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg 1861 gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct 1921 ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg 1981 gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag 2041 gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc 2101 gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga 2161 ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc 2221 ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat 2281 tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgacgcta 2341 gcccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta cccaccatga 2401 aggagaagag tgcttgtcct aaagatccag ccaaacctcc ggccaaggca caagttgtgg 2461 gatggccacc ggtgagatca taccggaaga acgtgatggt ttcctgccaa aaatcaagcg 2521 gtggcccgga ggcggcggcg ttcgtgaagg tatcaatgga cggagcaccg tacttgagga 2581 aaatcgattt gaggatgtat aaagctagca tggtgagcaa gggcgaggag ctgttcaccg 2641 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt 2701 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca 2761 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt 2821 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg 2881 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg 2941 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact 3001 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg 3061 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca 3121 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg 3181 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag 3241 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca 3301 ctctcggcat ggacgagctg tacaagggtg caggcgccca agcttcgaat tctgcagtcg 3361 accccaagaa gaaaagaaag gtcaggtagg gatccaccgg atctagataa ctgatcataa 3421 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc 3481 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata 3541 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 3601 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtcgat catattcaat 3661 aacccttaat ataacttcgt ataatgtatg ctatacgaag ttattaggtc tgaagaggag 3721 tttacgtcca gccaagctta ggatctcgac ctcgaaattc taccgggtag gggaggcgct 3781 tttcccaagg cagtctggag catgcgcttt agcagccccg ctgggcactt ggcgctacac 3841 aagtggcctc tggcctcgca cacattccac atccaccggt aggcgccaac cgactccgtt 3901 ctttggtggc cccttcgcgc caccttctac tcctccccta gtcaggaagt tcccccccgc 3961 cccgcagctc gcgtcgtgca ggacgtgaca aatggaagta gcacgtctca ctagtctcgt 4021 gcagatggac agcaccgctg agcaatggaa gcgggtaggc ctttggggca gcggccaata 4081 gcagctttgc tccttcgctt tctgggctca gaggctggga aggggtgggt ccgggggcgg 4141 gctcaggggc gggctcaggg gcggggcggg cgcccgaagg tcctccggag gcccggcatt 4201 ctgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct 4261 ttcgacctgc atccatctag atctcgagca gctgaagctt accgctagca tggatagatc 4321 cggaaagcct gaactcaccg cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag 4381 cgtctccgac ctgatgcagc tctcggaggg cgaagaatct cgtgctttca gcttcgatgt 4441 aggagggcgt ggatatgtcc tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg 4501 ttatgtttat cggcactttg catcggccgc gctcccgatt ccggaagtgc ttgacattgg 4561 ggaattcagc gagagcctga cctattgcat ctcccgccgt gcacagggtg tcacgttgca 4621 agacctgcct gaaaccgaac tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc 4681 gatcgctgcg gccgatctta gccagacgag cgggttcggc ccattcggac cgcaaggaat 4741 cggtcaatac actacatggc gtgatttcat atgcgcgatt gctgatcccc atgtgtatca 4801 ctggcaaact gtgatggacg acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct 4861 gatgctttgg gccgaggact gccccgaagt ccggcacctc gtgcacgcgg atttcggctc 4921 caacaatgtc ctgacggaca atggccgcat aacagcggtc attgactgga gcgaggcgat 4981 gttcggggat tcccaatacg aggtcgccaa catcttcttc tggaggccgt ggttggcttg 5041 tatggagcag cagacgcgct acttcgagcg gaggcatccg gagcttgcag gatcgccgcg 5101 gctccgggcg tatatgctcc gcattggtct tgaccaactc tatcagagct tggttgacgg 5161 caatttcgat gatgcagctt gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc 5221 cgggactgtc gggcgtacac aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg 5281 tgtagaagta ctcgccgata gtggaaaccg acgccccagc actcgtccga gggcaaagga 5341 ataggctagc cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca 5401 tgcatcgatg atatcagatc cccgggatgc agaaattgat gatctattaa acaataaaga 5461 tgtccactaa aatggaagtt tttcctgtca tactttgtta agaagggtga gaacagagta 5521 cctacatttt gaatggaagg attggagcta cgggggtggg ggtggggtgg gattagataa 5581 atgcctgctc tttactgaag gctctttact attgctttat gataatgttt catagttgga 5641 tatcataatt taaacaagca aaaccaaatt aagggccagc tcattcctcc cactcatgat 5701 ctatagatct atagatctct cgtgggatca ttgtttttct cttgattccc actttgtggt 5761 tctaagtact gtggtttcca aatgtgtcag tttcatagcc tgaagaacga gatcagcagc 5821 ctctgttcca catacacttc attctcagta ttgttttgcc aagttctaat tccatcagaa 5881 gctggtcgag atccggaacc cttaatataa cttcgtataa tgtatgctat acgaagttat 5941 taggtccctc gaagaggttc actagctagt taaaagtttt gttactttat agaagaaatt 6001 ttgagttttt gttttttttt aataaataaa taaacataaa taaattgttt gttgaattta 6061 ttattagtat gtaagtgtaa atataataaa acttaatatc tattcaaatt aataaataaa 6121 cctcgatata cagaccgata aaacacatgc gtcaatttta cgcatgatta tctttaacgt 6181 acgtcacaat atgattatct ttctagggtt aatctagtat acgcgtatgc ggccgcttaa 6241 ttaatccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc 6301 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 6361 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 6421 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 6481 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga 6541 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 6601 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 6661 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 6721 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 6781 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 6841 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 6901 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 6961 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 7021 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 7081 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 7141 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 7201 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 7261 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 7321 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat 7381 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga 7441 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg 7501 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga 7561 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc 7621 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac 7681 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc 7741 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc 7801 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc 7861 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt 7921 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc 7981 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg 8041 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag 8101 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat 8161 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc 8221 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa 8281 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta 8341 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa 8401 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac //