出品公司: | Novagen |
---|---|
载体名称: | pETcoco-1 |
质粒类型: | 原核表达载体 |
启动子: | T7/Lac |
载体大小: | 12474 bp |
载体抗性: | Chloramphenicol |
买家导航
pETcoco-1载体 pETcoco1质粒基本信息
pETcoco-1质粒图谱 pETcoco1载体图谱和pETcoco-1载体序列 pETcoco1质粒序列多克隆位点信息
pETcoco-1质粒 pETcoco1载体简介
ETcoco1表达式向量的设计允许向量的“按命令”放大 拷贝数从单个拷贝(每个细胞1-2个质粒拷贝)到中等拷贝(每个细胞20-50个)这个 利用F-上位机的oriS和repE元素以及抛物线分拆行列式,使向量保持在单拷贝状态介质拷贝状态是通过在oriV处使用RK-2衍生的trfA复制器获得的从单拷贝到介质的转换 复制状态是通过诱导trfa基因与阿拉伯糖的表达来实现的。这些载体的双重复制子特性促进了重组质粒的稳定性,并且 在单拷贝状态下的低基础表达水平,同时允许典型的pET载体的健壮的靶蛋白表达。唯一的限制站点显示在圆形地图上(如下)。这个 序列在+1处相对于T7转录开始编号。
pETcoco-1质粒序列 pETcoco1载体序列
LOCUS pETcoco-1 12474 bp ds-DNA circular SYN 23-OCT-2018 DEFINITION Bacterial vector with a chloramphenicol resistance marker that allows single-copy replication, on-demand amplification, and tightly regulated expression. ACCESSION . VERSION . KEYWORDS pETcoco-1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12474) AUTHORS MilliporeSigma (Novagen) TITLE Direct Submission JOURNAL Exported Wednesday, February 26, 2020 from SnapGene Viewer 3.1.4 http://www.snapgene.com COMMENT The copy number can be increased to 20-50 copies per cell by adding arabinose to the growth medium. FEATURES Location/Qualifiers source 1..12474 /organism="synthetic DNA construct" /lab_host="Escherichia coli" /mol_type="other DNA" protein_bind 3..27 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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)." RBS 58..63 /note="ribosome binding site" misc_feature 70..314 /note="MCS" /note="multiple cloning site" CDS 72..74 /codon_start=1 /product="start codon" /note="ATG" /translation="M" CDS 75..92 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS 102..146 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /note="S-Tag" /translation="KETAAAKFERQHMDS" CDS 162..176 /codon_start=1 /product="enterokinase recognition and cleavage site" /note="enterokinase site" /translation="DDDDK" CDS 249..281 /codon_start=1 /product="HSV (herpes simplex virus) epitope tag" /note="HSV tag" /translation="QPELAPEDPED" CDS 326..343 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" terminator 410..457 /note="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" CDS complement(831..1709) /codon_start=1 /gene="araC" /product="L-arabinose regulatory protein" /note="araC" /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE EKVNDVAVKLS" promoter 1855..2020 /gene="araBAD" /note="araBAD promoter" /note="promoter of the L-arabinose operon of E. coli; the araC regulatory gene is transcribed in the opposite direction" CDS 2080..3228 /codon_start=1 /product="trans-acting replication protein that binds to and activates oriV" /note="trfA" /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWLQVLEYAKRTPIGEPITFTF YELCQDLGWSINGRYYTKAEECLSRLQATAMDFTSDRVGHLESVSLLHRFRVLDRGKKT SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR" CDS complement(3396..4055) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /note="CmR" /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" promoter complement(4056..4158) /note="cat promoter" /note="promoter of the E. coli cat gene" rep_origin 5099..5713 /note="oriV" /note="origin of replication for the bacterial F plasmid" rep_origin 5813..6032 /note="ori2" /note="secondary origin of replication for the bacterial F plasmid; also known as oriS" CDS 6123..6878 /codon_start=1 /gene="repE" /product="replication initiation protein for the bacterial F plasmid" /note="repE" /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM RLSYIEKKKGRQTTHIVFSFRDITSMTTG" misc_feature 6881..7131 /gene="incC" /note="incC" /note="incompatibility region of the bacterial F plasmid" CDS 7457..8632 /codon_start=1 /gene="sopA" /product="partitioning protein for the bacterial F plasmid" /note="sopA" /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR" CDS 8632..9603 /codon_start=1 /gene="sopB" /product="partitioning protein for the bacterial F plasmid" /note="sopB" /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP" misc_feature 9676..10149 /gene="sopC" /note="sopC" /note="centromere-like partitioning region of the bacterial F plasmid" misc_feature 10409..10807 /note="cos" /note="lambda cos site; allows packaging into phage lambda particles" protein_bind 10825..10858 /bound_moiety="Cre recombinase" /note="loxP" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(10985..12067) /codon_start=1 /gene="lacI" /product="lac repressor" /note="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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(12068..12145) /gene="lacI" /note="lacI promoter" promoter 12458..2 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag 61 gagatataag catgcatcac catcaccatc acggtgctag caaagaaacc gctgctgcga 121 aatttgaacg ccagcacatg gactcgccac cgccaactgg tgatgacgac gacaagagcc 181 cgggcgatcg catcgatgac gtccaggcgc gccacgtgcg tacggtttcg aagcttgcgg 241 ccgcaagcca gccagaactc gctccggaag atcccgagga tctttaatta acctcaggat 301 ttaaataacc taggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa 361 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc 421 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg 481 attggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 541 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 601 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 661 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 721 tcacggccgc cgcaatgctt gcataatgtg gcctgtcaaa tggacgaagc agggattctg 781 caaaccctat gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac 841 ttgacggcta catcattcac tttttcttca caaccggcac gaaactcgct cgggctggcc 901 ccggtgcatt ttttaaatac tcgcgagaaa tagagttgat cgtcaaaacc aacattgcga 961 ccgacggtgg cgataggcat ccgggtagtg ctcaaaagca gcttcgcctg gctgatacgt 1021 tggtcctcgc gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga 1081 cgcgacggcg acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc 1141 aggtgatcgc tgatgtactg acaagcctcg cgtacccgat tatccatcgg cggatggagc 1201 gactcgttaa tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc 1261 agctccgaat agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg 1321 aaatgcggct ggtgcgcttc atccgggcga aagaaacccg tattggcaaa tattgacggc 1381 cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat 1441 tcgcgagcct ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata 1501 tcacccggtc ggcagacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg 1561 gtgagattga gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa 1621 ccgttggcct caatcggcgt taaacccgcc accagatggg cgttaaacga gtatcccggc 1681 agcaggggat cattttgcgc ttcagccata cttttcatac tcccaccatt cagagaagaa 1741 accaattgtc catattgcat cagacattgc cgtcactgcg tcttttactg gctcttctcg 1801 ctaacccaac cggtaacccc gcttattaaa agcattctgt aacaaagcgg gaccaaagcc 1861 atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca gaaaagtcca cattgattat 1921 ttgcacggcg tcacactttg ctatgccata gcatttttat ccataagatt agcggatcct 1981 acctgacgct ttttatcgca actctctact gtttctccat acccgttttt ttggatggag 2041 tgaaaggagg aattcggggc aatcccgcaa ggagggtgaa tgaatcggac gtttgaccgg 2101 aaggcataca ggcaagaact gatcgacgcg gggttttccg ccgaggatgc cgaaaccatc 2161 gcaagccgca ccgtcatgcg tgcgccccgc gaaaccttcc agtccgtcgg ctcgatggtc 2221 cagcaagcta cggccaagat cgagcgcgac agcgtgcaac tggctccccc tgccctgccc 2281 gcgccatcgg ccgccgtgga gcgttcgcgt cgtctcgaac aggaggcggc aggtttggcg 2341 aagtcgatga ccatcgacac gcgaggaact atgacgacca agaagcgaaa aaccgccggc 2401 gaggacctgg caaaacaggt cagcgaggcc aagcaggccg cgttgctgaa acacacgaag 2461 cagcagatca aggaaatgca gctttccttg ttcgatattg cgccgtggcc ggacacgatg 2521 cgagcgatgc caaacgacac ggcccgctct gccctgttca ccacgcgcaa caagaaaatc 2581 ccgcgcgagg cgctgcaaaa caaggtcatt ttccacgtca acaaggacgt gaagatcacc 2641 tacaccggcg tcgagctgcg ggccgacgat gacgaactgg tgtggctgca ggtgttggag 2701 tacgcgaagc gcacccctat cggcgagccg atcaccttca cgttctacga gctttgccag 2761 gacctgggct ggtcgatcaa tggccggtat tacacgaagg ccgaggaatg cctgtcgcgc 2821 ctacaggcga cggcgatgga cttcacgtcc gaccgcgttg ggcacctgga atcggtgtcg 2881 ctgctgcacc gcttccgcgt cctggaccgt ggcaagaaaa cgtcccgttg ccaggtcctg 2941 atcgacgagg aaatcgtcgt gctgtttgct ggcgaccact acacgaaatt catatgggag 3001 aagtaccgca agctgtcgcc gacggcccga cggatgttcg actatttcag ctcgcaccgg 3061 gagccgtacc cgctcaagct ggaaaccttc cgcctcatgt gcggatcgga ttccacccgc 3121 gtgaagaagt ggcgcgagca ggtcggcgaa gcctgcgaag agttgcgagg cagcggcctg 3181 gtggaacacg cctgggtcaa tgatgacctg gtgcattgca aacgctaggg ccttgtgggg 3241 tcagttccgg ctgggggttc agcagccacc gggccgtcga ccaattctca tgtttgacag 3301 cttatcatcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaacca 3361 ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg 3421 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga 3481 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg 3541 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa 3601 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag 3661 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa 3721 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg 3781 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat 3841 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc 3901 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag 3961 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata 4021 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat 4081 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa 4141 cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta 4201 tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat 4261 tcgcgataag ctcatggagc ggcgtaaccg tcgcacagga aggacagaga aagcgcggat 4321 ctgggaagtg acggacagaa cggtcaggac ctggattggg gaggcggttg ccgccgctgc 4381 tgctgacggt gtgacgttct ctgttccggt cacaccacat acgttccgcc attcctatgc 4441 gatgcacatg ctgtatgccg gtataccgct gaaagttctg caaagcctga tgggacataa 4501 gtccatcagt tcaacggaag tctacacgaa ggtttttgcg ctggatgtgg ctgcccggca 4561 ccgggtgcag tttgcgatgc cggagtctga tgcggttgcg atgctgaaac aattatcctg 4621 agaataaatg ccttggcctt tatatggaaa tgtggaactg agtggatatg ctgtttttgt 4681 ctgttaaaca gagaagctgg ctgttatcca ctgagaagcg aacgaaacag tcgggaaaat 4741 ctcccattat cgtagagatc cgcattatta atctcaggag cctgtgtagc gtttatagga 4801 agtagtgttc tgtcatgatg cctgcaagcg gtaacgaaaa cgatttgaat atgccttcag 4861 gaacaataga aatcttcgtg cggtgttacg ttgaagtgga gcggattatg tcagcaatgg 4921 acagaacaac ctaatgaaca cagaaccatg atgtggtctg tccttttaca gccagtagtg 4981 ctcgccgcag tcgagcgaca gggcgaagcc ctcgagatcc cctgccattt ttggggtgag 5041 gccgttcgcg gccgaggggc gcagcccctg gggggatggg aggcccgcgt tagcggccgg 5101 gagggttcga gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca 5161 gccctggtta aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta 5221 gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag 5281 cccctcaaat gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa 5341 ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact 5401 tatccccagg cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc 5461 gatttgcgag gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt 5521 caacgccgcg ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt 5581 gagggccaag ttttccgcga ggtatccaca acgccggcgg ccggccgcgg tgtctcgcac 5641 acggcttcga cggcgtttct ggcgcgtttg cagggccata gacggccgcc agcccagcgg 5701 cgagggcaac cagcccggtg agcgtcggaa aggggaattt cgagtgagcg aggaagcacc 5761 agggaacagc acttatatat tctgcttaca cacgatgcct gaaaaaactt cccttggggt 5821 tatccactta tccacgggga tatttttata attatttttt ttatagtttt tagatcttct 5881 tttttagagc gccttgtagg cctttatcca tgctggttct agagaaggtg ttgtgacaaa 5941 ttgccctttc agtgtgacaa atcaccctca aatgacagtc ctgtctgtga caaattgccc 6001 ttaaccctgt gacaaattgc cctcagaaga agctgttttt tcacaaagtt atccctgctt 6061 attgactctt ttttatttag tgtgacaatc taaaaacttg tcacacttca catggatctg 6121 tcatggcgga aacagcggtt atcaatcaca agaaacgtaa aaatagcccg cgaatcgtcc 6181 agtcaaacga cctcactgag gcggcatata gtctctcccg ggatcaaaaa cgtatgctgt 6241 atctgttcgt tgaccagatc agaaaatctg atggcaccct acaggaacat gacggtatct 6301 gcgagatcca tgttgctaaa tatgctgaaa tattcggatt gacctctgcg gaagccagta 6361 aggatatacg gcaggcattg aagagtttcg cggggaagga agtggttttt tatcgccctg 6421 aagaggatgc cggcgatgaa aaaggctatg aatcttttcc ttggtttatc aaacgtgcgc 6481 acagtccatc cagagggctt tacagtgtac atatcaaccc atatctcatt cccttcttta 6541 tcgggttaca gaaccggttt acgcagtttc ggcttagtga aacaaaagaa atcaccaatc 6601 cgtatgccat gcgtttatac gaatccctgt gtcagtatcg taagccggat ggctcaggca 6661 tcgtctctct gaaaatcgac tggatcatag agcgttacca gctgcctcaa agttaccagc 6721 gtatgcctga cttccgccgc cgcttcctgc aggtctgtgt taatgagatc aacagcagaa 6781 ctccaatgcg cctctcatac attgagaaaa agaaaggccg ccagacgact catatcgtat 6841 tttccttccg cgatatcact tccatgacga caggatagtc tgagggttat ctgtcacaga 6901 tttgagggtg gttcgtcaca tttgttctga cctactgagg gtaatttgtc acagttttgc 6961 tgtttccttc agcctgcatg gattttctca tactttttga actgtaattt ttaaggaagc 7021 caaatttgag ggcagtttgt cacagttgat ttccttctct ttcccttcgt catgtgacct 7081 gatatcgggg gttagttcgt catcattgat gagggttgat tatcacagtt tattactctg 7141 aattggctat ccgcgtgtgt acctctacct ggagtttttc ccacggtgga tatttcttct 7201 tgcgctgagc gtaagagcta tctgacagaa cagttcttct ttgcttcctc gccagttcgc 7261 tcgctatgct cggttacacg gctgcggcga gcgctagtga taataagtga ctgaggtatg 7321 tgctcttctt atctcctttt gtagtgttgc tcttatttta aacaactttg cggttttttg 7381 atgactttgc gattttgttg ttgctttgca gtaaattgca agatttaata aaaaaacgca 7441 aagcaatgat taaaggatgt tcagaatgaa actcatggaa acacttaacc agtgcataaa 7501 cgctggtcat gaaatgacga aggctatcgc cattgcacag tttaatgatg acagcccgga 7561 agcgaggaaa ataacccggc gctggagaat aggtgaagca gcggatttag ttggggtttc 7621 ttctcaggct atcagagatg ccgagaaagc agggcgacta ccgcacccgg atatggaaat 7681 tcgaggacgg gttgagcaac gtgttggtta tacaattgaa caaattaatc atatgcgtga 7741 tgtgtttggt acgcgattgc gacgtgctga agacgtattt ccaccggtga tcggggttgc 7801 tgcccataaa ggtggcgttt acaaaacctc agtttctgtt catcttgctc aggatctggc 7861 tctgaagggg ctacgtgttt tgctcgtgga aggtaacgac ccccagggaa cagcctcaat 7921 gtatcacgga tgggtaccag atcttcatat tcatgcagaa gacactctcc tgcctttcta 7981 tcttggggaa aaggacgatg tcacttatgc aataaagccc acttgctggc cggggcttga 8041 cattattcct tcctgtctgg ctctgcaccg tattgaaact gagttaatgg gcaaatttga 8101 tgaaggtaaa ctgcccaccg atccacacct gatgctccga ctggccattg aaactgttgc 8161 tcatgactat gatgtcatag ttattgacag cgcgcctaac ctgggtatcg gcacgattaa 8221 tgtcgtatgt gctgctgatg tgctgattgt tcccacgcct gctgagttgt ttgactacac 8281 ctccgcactg cagtttttcg atatgcttcg tgatctgctc aagaacgttg atcttaaagg 8341 gttcgagcct gatgtacgta ttttgcttac caaatacagc aatagtaatg gctctcagtc 8401 cccgtggatg gaggagcaaa ttcgggatgc ctggggaagc atggttctaa aaaatgttgt 8461 acgtgaaacg gatgaagttg gtaaaggtca gatccggatg agaactgttt ttgaacaggc 8521 cattgatcaa cgctcttcaa ctggtgcctg gagaaatgct ctttctattt gggaacctgt 8581 ctgcaatgaa attttcgatc gtctgattaa accacgctgg gagattagat aatgaagcgt 8641 gcgcctgtta ttccaaaaca tacgctcaat actcaaccgg ttgaagatac ttcgttatcg 8701 acaccagctg ccccgatggt ggattcgtta attgcgcgcg taggagtaat ggctcgcggt 8761 aatgccatta ctttgcctgt atgtggtcgg gatgtgaagt ttactcttga agtgctccgg 8821 ggtgatagtg ttgagaagac ctctcgggta tggtcaggta atgaacgtga ccaggagctg 8881 cttactgagg acgcactgga tgatctcatc ccttcttttc tactgactgg tcaacagaca 8941 ccggcgttcg gtcgaagagt atctggtgtc atagaaattg ccgatgggag tcgccgtcgt 9001 aaagctgctg cacttaccga aagtgattat cgtgttctgg ttggcgagct ggatgatgag 9061 cagatggctg cattatccag attgggtaac gattatcgcc caacaagtgc ttatgaacgt 9121 ggtcagcgtt atgcaagccg attgcagaat gaatttgctg gaaatatttc tgcgctggct 9181 gatgcggaaa atatttcacg taagattatt acccgctgta tcaacaccgc caaattgcct 9241 aaatcagttg ttgctctttt ttctcacccc ggtgaactat ctgcccggtc aggtgatgca 9301 cttcaaaaag cctttacaga taaagaggaa ttacttaagc agcaggcatc taaccttcat 9361 gagcagaaaa aagctggggt gatatttgaa gctgaagaag ttatcactct tttaacttct 9421 gtgcttaaaa cgtcatctgc atcaagaact agtttaagct cacgacatca gtttgctcct 9481 ggagcgacag tattgtataa gggcgataaa atggtgctta acctggacag gtctcgtgtt 9541 ccaactgagt gtatagagaa aattgaggcc attcttaagg aacttgaaaa gccagcaccc 9601 tgatgcgacc acgttttagt ctacgtttat ctgtctttac ttaatgtcct ttgttacagg 9661 ccagaaagca taactggcct gaatattctc tctgggccca ctgttccact tgtatcgtcg 9721 gtctgataat cagactggga ccacggtccc actcgtatcg tcggtctgat tattagtctg 9781 ggaccacggt cccactcgta tcgtcggtct gattattagt ctgggaccac ggtcccactc 9841 gtatcgtcgg tctgataatc agactgggac cacggtccca ctcgtatcgt cggtctgatt 9901 attagtctgg gaccatggtc ccactcgtat cgtcggtctg attattagtc tgggaccacg 9961 gtcccactcg tatcgtcggt ctgattatta gtctggaacc acggtcccac tcgtatcgtc 10021 ggtctgatta ttagtctggg accacggtcc cactcgtatc gtcggtctga ttattagtct 10081 gggaccacga tcccactcgt gttgtcggtc tgattatcgg tctgggacca cggtcccact 10141 tgtattgtcg atcagactat cagcgtgaga ctacgattcc atcaatgcct gtcaagggca 10201 agtattgaca tgtcgtcgta acctgtagaa cggagtaacc tcggtgtgcg gttgtatgcc 10261 tgctgtggat tgctgctgtg tcctgcttat ccacaacatt ttgcgcacgg ttatgtggac 10321 aaaatacctg gttacccagg ccgtgccggc acgttaaccg ggctgcatcc gatgcaagtg 10381 tgtcgctgtc gacgagctcg cgagctcgga catgaggttg ccccgtattc agtgtcgctg 10441 atttgtattg tctgaagttg tttttacgtt aagttgatgc agatcaatta atacgatacc 10501 tgcgtcataa ttgattattt gacgtggttt gatggcctcc acgcacgttg tgatatgtag 10561 atgataatca ttatcacttt acgggtcctt tccggtgatc cgacaggtta cggggcggcg 10621 acctcgcggg ttttcgctat ttatgaaaat tttccggttt aaggcgtttc cgttcttctt 10681 cgtcataact taatgttttt atttaaaata ccctctgaaa agaaaggaaa cgacaggtgc 10741 tgaaagcgag ctttttggcc tctgtcgttt cctttctctg tttttgtccg tggaatgaac 10801 aatggaagtc cgagctcatc gctaataact tcgtatagca tacattatac gaagttatat 10861 tcgatgcggc ctagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct 10921 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt 10981 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg 11041 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca 11101 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc 11161 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga 11221 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc 11281 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca 11341 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca 11401 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt 11461 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg 11521 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat 11581 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa 11641 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga 11701 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc 11761 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa 11821 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca 11881 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg 11941 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca 12001 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg 12061 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa 12121 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc 12181 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt 12241 gcatgtaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg 12301 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg 12361 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg 12421 gcgtagagga tcgagatcga tctcgatccc gcgaaattaa tacgactcac tata //
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