LOCUS Exported 6972 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pQexp SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6972) AUTHORS Tolonen AC, Chilaka AC, Church GM TITLE Targeted gene inactivation in Clostridium phytofermentans shows that cellulose degradation requires the family 9 hydrolase Cphy3367. JOURNAL Mol Microbiol. 2009 Dec . 74(6):1300-13. PUBMED 19775243 REFERENCE 2 (bases 1 to 6972) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6972 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 387..975 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 876..895 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1129..1146 /label=L4440 /note="L4440 vector, forward primer" protein_bind 1263..1284 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 1299..1329 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1337..1353 /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)." primer_bind 1342..1364 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 1361..1377 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 1361..1377 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1860..1877) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(1860..1876) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1869..1891) /label=M13/pUC Forward /note="In lacZ gene" CDS complement(2195..2566) /codon_start=1 /gene="traJ" /product="oriT-recognizing protein" /label=traJ /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE KQDELGKVMMGVVRPRAEP" oriT 2599..2708 /note="incP origin of transfer" ORIGIN 1 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 61 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 121 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 181 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 241 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 301 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 361 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 421 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 481 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta 541 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 601 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 661 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 721 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 781 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 841 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 901 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 961 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 1021 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc 1081 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg 1141 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt 1201 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca 1261 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct 1321 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat 1381 gattacaagc ttgcatgcct gcagtttggt caacatttaa cctctaatct tctagttcat 1441 ttatatgtcc atgaattgaa tttcatacat agttttcaat tgtatggatg tagtatgaca 1501 gaaaatcaaa aaatcatact tattatattt aggtagttac tgttaatttt tagtatgtta 1561 atttctaatc tgagttcttt ccgtattata ataagtgtaa agatcacacc atatttttac 1621 agtttttaca cgtttcatta ataatttctt atatttcttt atttgtttgt aaaatttact 1681 taaatttcgc cagaaaacaa aagaaagcct ttactaatta atagtttagt gatactcttt 1741 tatgtaggta ttttttaaaa tacattaaac ctaggtaatt gaggaaagtt acaattacca 1801 ttatataagg aggatattca aaccatatgg gatccccggg taccgagctc gaattcggca 1861 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc 1921 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc 1981 ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt 2041 acgcatctgt gcggtatttc acaccggatc ggtcttgcct tgctcgtcgg tgatgtactt 2101 caccagctcc gcgaagtcgc tcttcttgat ggagcgcatg gggacgtgct tggcaatcac 2161 gcgcaccccc cggccgtttt agcggctaaa aaagtcatgg ctctgccctc gggcggacca 2221 cgcccatcat gaccttgcca agctcgtcct gcttctcttc gatcttcgcc agcagggcga 2281 ggatcgtggc atcaccgaac cgcgccgtgc gcgggtcgtc ggtgagccag agtttcagca 2341 ggccgcccag gcggcccagg tcgccattga tgcgggccag ctcgcggacg tgctcatagt 2401 ccacgacgcc cgtgattttg tagccctggc cgacggccag caggtaggcc gacaggctca 2461 tgccggccgc cgccgccttt tcctcaatcg ctcttcgttc gtctggaagg cagtacacct 2521 tgataggtgg gctgcccttc ctggttggct tggtttcatc agccatccgc ttgccctcat 2581 ctgttacgcc ggcggtagcc ggccagcctc gcagagcagg attcccgttg agcaccgcca 2641 ggtgcgaata agggacagtg aagaaggaac acccgctcgc gggtgggcct acttcaccta 2701 tcctgcccgg ctgacgccgt tggatacacc aaggaaagtc tacacgaacc ctttggcaaa 2761 atcctgtata tcgtgcgaaa aaggatggat ataccgaaaa aatcgctata atgaccccga 2821 agcagggtta tgcagcggaa aagatccgtc gaccttagaa gcaaacttaa gagtgtgttg 2881 atagtgcagt atcttaaaat tttgtgtata ataggaattg aagttaaatt agatgctaaa 2941 aatttgtaat taagaaggag ggattcgtca tgttggtatt ccaaatgcgt aatgtagata 3001 aaacatctac tgttttgaaa cagactaaaa acagtgatta cgcagataaa taaatacgtt 3061 agattaattc ctaccagtga ctaatcttat gactttttaa acagataact aaaattacaa 3121 acaaatcgtt taacttctgt atttatttac agatgtaatc acttcaggag taattacatg 3181 aacaaaaata taaaatattc tcaaaacttt ttaacgagtg aaaaagtact caaccaaata 3241 ataaaacaat tgaatttaaa agaaaccgat accgtttacg aaattggaac aggtaaaggg 3301 catttaacga cgaaactggc taaaataagt aaacaggtaa cgtctattga attagacagt 3361 catctattca acttatcgtc agaaaaatta aaactgaaca ttcgtgtcac tttaattcac 3421 caagatattc tacagtttca attccctaac aaacagaggt ataaaattgt tgggagtatt 3481 ccttaccatt taagcacaca aattattaaa aaagtggttt ttgaaagcca tgcgtctgac 3541 atctatctga ttgttgaaga aggattctac aagcgtacct tggatattca ccgaacacta 3601 gggttgctct tgcacactca agtctcgatt cagcaattgc ttaagctgcc agcggaatgc 3661 tttcatccta aaccaaaagt aaacagtgtc ttaataaaac ttacccgcca taccacagat 3721 gttccagata aatattggaa gctatatacg tactttgttt caaaatgggt caatcgagaa 3781 tatcgtcaac tgtttactaa aaatcagttt catcaagcaa tgaaacacgc caaagtaaac 3841 aatttaagta ccattactta tgagcaagta ttgtctattt ttaatagtta tctattattt 3901 aacgggagga aataattcta tgagtcgctt ttttaaattt ggaaagttac acgttactaa 3961 agggaatgga gataaattat tagatatact actgacagct tccaagaagc taaagaggtc 4021 cctagcgcct acggggaatt tgtaaatcaa aagaaaaatt ctcccaagaa ttagaaatga 4081 gtagatcaaa ttattcacga atagaatcag gaaaatcaga tccaaccata aaaacactag 4141 aacaaattgc aaagttaact aactcaacgc tagtagtgga tttaatccca aatgagccaa 4201 cagaaccaga accagaaaca gaatcagaac aagtaacatt ggatttagaa atggaagaag 4261 aaaaaagcaa tgacttcgtg tgaataatgc acgaaatcgt tgcttatttt tttttaaaag 4321 cggtatacta gatataacga aacaacgaac tgaatagaaa cgaaaaaaga gccatgacac 4381 atttataaaa tgtttgacga cattttataa atgcatagcc cgataagatt gccaaaccaa 4441 cgcttatcag ttagtcagat gaactcttcc ctcgtaagaa gttatttaat taactttgtt 4501 tgaagacggt atataaccgt actatcatta tatagggaaa tcagagagtt ttcaagtatc 4561 taagctactg aatttaagaa ttgttaagca atcaatcgga aatcgtttga ttgctttttt 4621 tgtattcatt tatagaaggt ggagtttgta tgaatcatga tgaatgtaaa acttatataa 4681 aaaatagttt attggagata agaaaattag caaatatcta tacactagaa acgtttaaga 4741 aagagttaga aaagagaaat atctacttag aaacaaaatc agataagtat ttttcttcgg 4801 agggggaaga ttatatatat aagttaatag aaaataacaa aataatttat tcgattagtg 4861 gaaaaaaatt gacttataaa ggaaaaaaat ctttttcaaa acatgcaata ttgaaacagt 4921 tgaatgaaaa agcaaaccaa gttaattaaa caacctattt tataggattt ataggaaagg 4981 agaacagctg aatgaatatc ccttttgttg tagaaactgt gcttcatgac ggcttgttaa 5041 agtacaaatt taaaaatagt aaaattcgct caatcactac caagccaggt aaaagcaaag 5101 gggctatttt tgcgtatcgc tcaaaatcaa gcatgattgg cggtcgtggt gttgttctga 5161 cttccgagga agcgattcaa gaaaatcaag atacatttac acattggaca cccaacgttt 5221 atcgttatgg aacgtatgca gacgaaaacc gttcatacac gaaaggacat tctgaaaaca 5281 atttaagaca aatcaatacc ttctttattg attttgatat tcacacggca aaagaaacta 5341 tttcagcaag cgatatttta acaaccgcta ttgatttagg ttttatgcct actatgatta 5401 tcaaatctga taaaggttat caagcatatt ttgttttaga aacgccagtc tatgtgactt 5461 caaaatcaga atttaaatct gtcaaagcag ccaaaataat ttcgcaaaat atccgagaat 5521 attttggaaa gtctttgcca gttgatctaa cgtgtaatca ttttggtatt gctcgcatac 5581 caagaacgga caatgtagaa ttttttgatc ctaattaccg ttattctttc aaagaatggc 5641 aagattggtc tttcaaacaa acagataata agggctttac tcgttcaagt ctaacggttt 5701 taagcggtac agaaggcaaa aaacaagtag atgaaccctg gtttaatctc ttattgcacg 5761 aaacgaaatt ttcaggagaa aagggtttaa tagggcgtaa taacgtcatg tttaccctct 5821 ctttagccta ctttagttca ggctattcaa tcgaaacgtg cgaatataat atgtttgagt 5881 ttaataatcg attagatcaa cccttagaag aaaaagaagt aatcaaaatt gttagaagtg 5941 cctattcaga aaactatcaa ggggctaata gggaatacat taccattctt tgcaaagctt 6001 gggtatcaag tgatttaacc agtaaagatt tatttgtccg tcaagggtgg tttaaattca 6061 agaaaaaaag aagcgaacgt caacgtgttc atttgtcaga atggaaagaa gatttaatgg 6121 cttatattag cgaaaaaagc gatgtataca agccttattt agtgacgacc aaaaaagaga 6181 ttagagaagt gctaggcatt cctgaacgga cattagataa attgctgaag gtactgaagg 6241 cgaatcagga aattttcttt aagattaaac caggaagaaa tggtggcatt caacttgcta 6301 gtgttaaatc attgttgcta tcgatcatta aagtaaaaaa agaagaaaaa gaaagctata 6361 taaaggcgct gacaaattct tttgacttag agcatacatt cattcaagag actttaaaca 6421 agctagcaga acgccctaaa acggacacac aactcgattt gtttagctat gatacaggct 6481 gaaaataaaa cccgcactat gccattacat ttatatctat gatacgtgtt tgttttttct 6541 ttgctgttta gcgaatgatt agcagaaata tacagagtaa gattttaatt aattattagg 6601 gggagaagga gagagtagcc cgaaaacttt tagttggctt ggactgaacg aagtgaggga 6661 aaggctacta aaacgtcgag gggcagtgag agcgaagcga acacttgatt ttttaatttt 6721 ctatctttta taggtcatta gagtatactt atttgtccta taaactattt agcagcataa 6781 tagatttatt gaataggtca tttaagttga gcatattaga ggaggaaaat cttggagaaa 6841 tatttgaaga acccgattac atggattgga ttagttcttg tggttacgtg gtttttaact 6901 aaaagtagtg aatttttgat ttttggtgtg tgtgtcttgt tgttagtatt tgctagtcaa 6961 agtgattaaa ta //