质粒名称: | vYM112_PB-pEF1a-optiPIN2-HA(Hygro) |
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出品公司: | Addgene |
目录编号: | 160045 |
存储实验室: | Michael Elowitz |
载体骨架基本信息 |
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载体名称: | PiggyBac |
载体出品公司: | SYSTEM BIOSCIENCES |
载体类型: | 哺乳动物表达,合成生物学 |
筛选标记: | 潮霉素 |
载体抗性: | 氨苄霉素,100 μg/mL |
生长温度: | 30 ℃ |
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: | PIN2 |
物种来源: | A. thaliana (mustard weed) |
插入大小: | 1941 bp |
基因突变: | codon optimized |
GenBank编号 | AF086907.1 |
启动子: | pEF1s |
标签/融合蛋白: | HA(插入件上的C端子) |
克隆方法: | Gibson克隆 |
5’测序引物 | taaggtagcgaaggtctgCGTAATG |
3’测序引物 | agggataacagggtaatgaggtTTAtgcgtagtctggtacgtcgtacggatagccgc |
文献引用方法 |
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材料和方法部分: | vYM112_PB-pEF1a-optiPIN2-HA(Hygro) was a gift from Michael Elowitz (Addgene plasmid # 160045 ; http://n2t.net/addgene:160045 ; RRID:Addgene_160045) |
参考文献部分: | Synthetic mammalian signaling circuits for robust cell population control. Ma Y, Budde MW, Mayalu MN, Zhu J, Lu AC, Murray RM, Elowitz MB. Cell. 2022 Mar 17;185(6):967-979.e12. doi: 10.1016/j.cell.2022.01.026. Epub 2022 Mar 1. 10.1016/j.cell.2022.01.026 PubMed 35235768 |
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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 vYM112_PB-pEF1a-optiPIN2-HA(Hygro) 9590 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION For expressing the active auxin pump PIN2. ACCESSION . VERSION . KEYWORDS vYM112_PB-pEF1a-optiPIN2-HA(Hygro) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9590) TITLE Synthetic mammalian signaling circuits for robust cell population control JOURNAL Unpublished REFERENCE 2 (bases 1 to 9590) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9590 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(59..76) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(230..818) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(310..329) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(989..1849) /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 1612..1631 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1850..1954) /gene="bla" /label=AmpR promoter rep_origin complement(1980..2435) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2117..2138) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2329..2348 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2562..2584 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2576..2593 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2577..2593 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" repeat_region 2723..2785 /label=piggyBac right (3') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" promoter 3228..4406 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 3459..4397 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 4354..4374 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 6418..6444 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" primer_bind complement(6472..6489) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6478..6685 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(6718..6738) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6723..7052 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6903..7038 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6965..6984 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 7101..8126 /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" primer_bind complement(8186..8206) /label=WPRE-R /note="WPRE, reverse primer" polyA_signal 8249..8370 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8286..8305) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8340..8359 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" repeat_region 8893..8927 /label=piggyBac left (5') inverted repeat /note="piggyBac transposon-specific inverted terminal repeat sequence (ITR)" primer_bind complement(9418..9434) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9418..9434) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(9431..9453) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 9442..9458 /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(9466..9496) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9511..9532 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct 61 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa 121 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa 181 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc 241 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga 301 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc 361 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt 421 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct 481 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg 541 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta 601 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct 661 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa 721 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt 781 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta 841 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat 901 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa 961 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct 1021 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta 1081 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gatccacgct 1141 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg 1201 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa 1261 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt 1321 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta 1381 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca 1441 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta 1501 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct 1561 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg 1621 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac 1681 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact 1741 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa 1801 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt 1861 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat 1921 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta 1981 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt 2041 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat 2101 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa 2161 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta 2221 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc 2281 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc 2341 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac 2401 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa 2461 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg 2521 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa 2581 aacgacggcc agtgagcgcg cctcgttcat tcacgttttt gaacccgtgg aggacgggca 2641 gactcgcggt gcaaatgtgt tttacagcgt gatggagcag atgaagatgc tcgacacgct 2701 gcagaacacg cagctagatt aaccctagaa agataatcat attgtgacgt acgttaaaga 2761 taatcatgtg taaaattgac gcatgtgttt tatcggtctg tatatcgagg tttatttatt 2821 aatttgaata gatattaagt tttattatat ttacacttac atactaataa taaattcaac 2881 aaacaattta tttatgttta tttatttatt aaaaaaaaca aaaactcaaa atttcttcta 2941 taaagtaaca aaacttttat gagggacagc ccccccccaa agcccccagg gatgtaatta 3001 cgtccctccc ccgctagggg gcagcagcga gccgcccggg gctccgctcc ggtccggcgc 3061 tccccccgca tccccgagcc ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca 3121 cgggatcgct ttcctctgaa cgcttctcgc tgctctttga gcctgcagac acctgggggg 3181 atacggggaa aaggcctcca cggccagcac ctgcgtgacg tacgtgaggc tccggtgccc 3241 gtcagtgggc agagcgcaca tcgcccacag tccccgagaa gttgggggga ggggtcggca 3301 attgaaccgg tgcctagaga aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact 3361 ggctccgcct ttttcccgag ggtgggggag aaccgtatat aagtgcagta gtcgccgtga 3421 acgttctttt tcgcaacggg tttgccgcca gaacacaggt aagtgccgtg tgtggttccc 3481 gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc ttgaattact tccacctggc 3541 tgcagtacgt gattcttgat cccgagcttc gggttggaag tgggtgggag agttcgaggc 3601 cttgcgctta aggagcccct tcgcctcgtg cttgagttga ggcctggcct gggcgctggg 3661 gccgccgcgt gcgaatctgg tggcaccttc gcgcctgtct cgctgctttc gataagtctc 3721 tagccattta aaatttttga tgacctgctg cgacgctttt tttctggcaa gatagtcttg 3781 taaatgcggg ccaagatctg cacactggta tttcggtttt tggggccgcg ggcggcgacg 3841 gggcccgtgc gtcccagcgc acatgttcgg cgaggcgggg cctgcgagcg cggccaccga 3901 gaatcggacg ggggtagtct caagctggcc ggcctgctct ggtgcctggc ctcgcgccgc 3961 cgtgtatcgc cccgccctgg gcggcaaggc tggcccggtc ggcaccagtt gcgtgagcgg 4021 aaagatggcc gcttcccggc cctgctgcag ggagctcaaa atggaggacg cggcgctcgg 4081 gagagcgggc gggtgagtca cccacacaaa ggaaaagggc ctttccgtcc tcagccgtcg 4141 cttcatgtga ctccacggag taccgggcgc cgtccaggca cctcgattag ttctcgagct 4201 tttggagtac gtcgtcttta ggttgggggg aggggtttta tgcgatggag tttccccaca 4261 ctgagtgggt ggagactgaa gttaggccag cttggcactt gatgtaattc tccttggaat 4321 ttgccctttt tgagtttgga tcttggttca ttctcaagcc tcagacagtg gttcaaagtt 4381 tttttcttcc atttcaggtg tcgtgaacct tcacgcaggt ggaattctaa ctataacggt 4441 cctaaggtag cgaaggtctg cgtaatgatc actggcaaag atatgtacga tgtccttgct 4501 gcaatggtac cactctatgt agcaatgatt cttgcttatg gcagtgtccg ctggtggggc 4561 atatttacac ccgatcagtg cagcgggatc aatcggttcg ttgccgtttt tgcagtaccc 4621 ctcctttcat tccacttcat ttcaagcaat gatccttacg caatgaatta tcactttctc 4681 gctgcagaca gtcttcagaa agtggtaatc ttggcagcac tctttctttg gcaggccttc 4741 tctagaagag gctctttgga gtggatgatt accctcttct ctctttctac tctccccaac 4801 acattggtga tgggcatacc tcttcttaga gcaatgtacg gggatttctc tggcaacctc 4861 atggtccaga tcgtggtgct tcagtctatc atatggtata cccttatgct gttccttttt 4921 gagttccgcg gggcaaaact gctcataagt gaacaattcc cagagaccgc cgggtctatc 4981 accagcttcc gggtggactc tgacgtaata agcctgaacg ggagagaacc ccttcaaaca 5041 gacgctgaaa taggagacga tggtaagttg catgtcgtag tacggaggtc aagcgctgct 5101 tcctcaatga tatctagttt caacaaatcc catggaggcg gccttaattc tagtatgata 5161 acaccaagag ccagcaatct cacaggcgta gaaatttact cagtgcagtc aagtagggaa 5221 ccaaccccac gcgcctcctc ttttaatcaa actgactttt atgccatgtt taacgcaagt 5281 aaggccccta gccctagaca cggctatact aatagttacg ggggcgctgg ggctggccca 5341 ggaggtgatg tctacagttt gcaatcaagt aagggggtaa ctcctcgcac ttcaaacttt 5401 gacgaggaag taatgaagac tgccaagaaa gcaggaaggg gcggacgcag tatgtcaggg 5461 gaactgtaca acaataactc agttccttct tatccaccac ctaacccaat gtttacaggc 5521 tcaacctctg gtgcatcagg cgtaaagaaa aaggaaagcg gtggaggcgg ctccggaggt 5581 ggcgtcggcg taggaggcca gaacaaagag atgaacatgt ttgtttggag ttccagtgct 5641 agcccagtca gtgaggctaa tgccaaaaac gcaatgaccc ggggttctag cacagatgtt 5701 tctaccgacc caaaggtgtc tatccctcca catgataatt tggcaaccaa agccatgcaa 5761 aacctcatcg aaaatatgag tcctggcaga aagggccatg tggagatgga tcaagatgga 5821 aacaatggtg gtaaaagccc ttatatgggg aaaaaaggct cagacgtcga agatggcgga 5881 cctggaccta gaaaacagca gatgccccca gcttccgtaa tgacacgact catccttatt 5941 atggtgtggc ggaaactcat acgcaatcca aacacctaca gcagcttgtt tgggctcgcc 6001 tggtctcttg tatcattcaa gtggaacatt aaaatgccaa caataatgag cgggtctatc 6061 tctattcttt cagatgctgg tttgggtatg gccatgttta gtttgggcct gttcatggct 6121 ctccaaccca agataattgc ttgtggaaag tctgtagccg gctttgctat ggccgtgcgg 6181 ttcctcacag gacctgccgt aatagcagct acttcaatcg ccataggtat caggggtgac 6241 ctgcttcaca ttgctattgt tcaagccgcc ttgcctcagg gcatcgtacc tttcgtcttt 6301 gctaaggaat ataacgttca tcctgacata ctctccaccg ctgtcatctt cgggatgttg 6361 gttgcattgc cagtaacagt tctgtactac gttcttcttg gccttggcgg aagcggctat 6421 ccgtacgacg taccagacta cgcataaacc tcattaccct gttatcccta gcctcgactg 6481 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg 6541 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga 6601 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg 6661 aagacaatag caggcatgct ggggacgcgg ccgcaattta acgcgaatta attctgtgga 6721 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 6781 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca 6841 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc 6901 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt 6961 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag 7021 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt 7081 tcggatctga tcagcacgtg atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt 7141 ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat 7201 ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg 7261 ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga 7321 ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc 7381 gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc 7441 cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg 7501 gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga 7561 ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg 7621 tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc 7681 tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg 7741 tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct 7801 tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc 7861 cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac 7921 tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg 7981 acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg 8041 cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca 8101 gcactcgtcc gagggcaaag gaataggtcg acaatcaacc tctggattac aaaatttgtg 8161 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt 8221 taatgccttt gtatcatccc tagaattcaa cttgtttatt gcagcttata atggttacaa 8281 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 8341 tggtttgtcc aaactcatca atgtatctta ggcccattgg tatggctttt tccccgtatc 8401 cccccaggtg tctgcaggct caaagagcag cgagaagcgt tcagaggaaa gcgatcccgt 8461 gccaccttcc ccgtgcccgg gctgtccccg cacgctgccg gctcggggat gcggggggag 8521 cgccggaccg gagcggagcc ccgggcggct cgctgctgcc ccctagcggg ggagggacgt 8581 aattacatcc ctgggggctt tggggggggg ctgtccctga tatctataac aagaaaatat 8641 atatataata agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt 8701 taaatcttaa aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat 8761 agttcaaaat cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg 8821 cgactgagat gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat 8881 atttcaagaa tgcatgcgtc aattttacgc agactatctt tctagggtta atctagctgc 8941 atcaggatca tatcgtcggg tcttttttcc ggctcagtca tcgcccaagc tggcgctatc 9001 tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga ggttgaagcg gcatggaaag 9061 agtttgccga ggatgactgc tgctgcattg acgttgagcg aaaacgcacg tttaccatga 9121 tgattcggga aggtgtggcc atgcacgcct ttaacggtga actgttcgtt caggccacct 9181 gggataccag ttcgtcgcgg cttttccgga cacagttccg gatggtcagc ccgaagcgca 9241 tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg tgccggtgtg cagattaatg 9301 acagcggtgc ggcgctggga tattacgtca gcgaggacgg gtatcctggc tggatgccgc 9361 agaaatggac atggataccc cgtgagttac ccggcgggcg cgcttggcgt aatcatggtc 9421 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg 9481 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt 9541 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt //