pETcoco-1

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pETcoco-1载体 pETcoco1质粒基本信息

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

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载体图谱
pETcoco-1载体图谱1

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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