LOCUS Exported 10168 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pQint SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10168) 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 10168) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10168 /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 2659..2678 /label=T7 /note="T7 promoter, forward primer" promoter 2659..2677 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" RBS 2840..2862 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" primer_bind complement(4876..4892) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5056..5072) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS complement(5391..5762) /codon_start=1 /gene="traJ" /product="oriT-recognizing protein" /label=traJ /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE KQDELGKVMMGVVRPRAEP" oriT 5795..5904 /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 gatccccagc ttataattat ccttagattt 1861 catcaatgtg cgcccagata gggtgttaag tcaagtagtt taaggtacta ctctgtaaga 1921 taacacagaa aacagccaac ctaaccgaaa agcgaaagct gatacgggaa cagagcacgg 1981 ttggaaagcg atgagttacc taaagacaat cgggtacgac tgagtcgcaa tgttaatcag 2041 atataaggta taagttgtgt ttactgaacg caagtttcta atttcgatta aatctcgata 2101 gaggaaagtg tctgaaacct ctagtacaaa gaaaggtaag ttagcattga tgacttatct 2161 gttatcacca catttgtaca atctgtagga gaacctatgg gaacgaaacg aaagcgatgc 2221 cgagaatctg aatttaccaa gacttaacac taactgggga taccctaaac aagaatgcct 2281 aatagaaagg aggaaaaagg ctatagcact agagcttgaa aatcttgcaa gggtacggag 2341 tactcgtagt agtctgagaa gggtaacgcc ctttacatgg caaaggggta cagttattgt 2401 gtactaaaat taaaaattga ttagggagga aaacctcaaa atgaaaccaa caatggcaat 2461 tttagaaaga atcagtaaaa attcacaaga aaatatagac gaagttttta caagacttta 2521 tcgttatctt ttacgtccag atatttatta cgtggcgacg cgttgggaaa tggcaatgat 2581 agcgaaacaa cgtaaaactc ttgttgtatg ctttcattgt catcgtcacg tgattcataa 2641 acacaagtga atgtcgacta atacgactca ctatagggtc gacagtgaat ttttacgaac 2701 gaacaataac agagccgtat actccgagag gggtacgtac ggttcccgaa gagggtggtg 2761 caaaccagtc acagtaatgt gaacaaggcg gtacctccct acttcaccat atcattttct 2821 gcagccccct agaaataatt ttgtttaact ttaagaagga gatatacata tatggctaga 2881 tcgtccattc cgacagcatc gccagtcact atggcgtgct gctagcgcta tatgcgttga 2941 tgcaatttct atgcactcgt agtagtctga gaagggtaac gccctttaca tggcaaaggg 3001 gtacagttat tgtgtactaa aattaaaaat tgattaggga ggaaaacctc aaaatgaaac 3061 caacaatggc aattttagaa agaatcagta aaaattcaca agaaaatata gacgaagttt 3121 ttacaagact ttatcgttat cttttacgtc cagatattta ttacgtggcg tatcaaaatt 3181 tatattccaa taaaggagct tccacaaaag gaatattaga tgatacagcg gatggcttta 3241 gtgaagaaaa aataaaaaag attattcaat ctttaaaaga cggaacttac tatcctcaac 3301 ctgtacgaag aatgtatatt gcaaaaaaga attctaaaaa gatgagacct ttaggaattc 3361 caactttcac agataaattg atccaagaag ctgtgagaat aattcttgaa tctatctatg 3421 aaccggtatt cgaagatgtg tctcacggtt ttagacctca acgaagctgt cacacagctt 3481 tgaaaacaat caaaagagag tttggcggcg caagatggtt tgtggaggga gatataaaag 3541 gctgcttcga taatatagac cacgttacac tcattggact catcaatctt aaaatcaaag 3601 atatgaaaat gagccaattg atttataaat ttctaaaagc aggttatctg gaaaactggc 3661 agtatcacaa aacttacagc ggaacacctc aaggtggaat tctatctcct cttttggcca 3721 acatctatct tcatgaattg gataagtttg ttttacaact caaaatgaag tttgaccgag 3781 aaagtccaga aagaataaca cctgaatatc gggagctcca caatgagata aaaagaattt 3841 ctcaccgtct caagaagttg gagggtgaag aaaaagctaa agttctttta gaatatcaag 3901 aaaaacgtaa aagattaccc acactcccct gtacctcaca gacaaataaa gtattgaaat 3961 acgtccggta tgcggacgac ttcattatct ctgttaaagg aagcaaagag gactgtcaat 4021 ggataaaaga acaattaaaa ctttttattc ataacaagct aaaaatggaa ttgagtgaag 4081 aaaaaacact catcacacat agcagtcaac ccgctcgttt tctgggatat gatatacgag 4141 taaggagatc tggaacgata aaacgatctg gtaaagtcaa aaagagaaca ctcaatggga 4201 gtgtagaact ccttattcct cttcaagaca aaattcgtca atttattttt gacaagaaaa 4261 tagctatcca aaagaaagat agctcatggt ttccagttca caggaaatat cttattcgtt 4321 caacagactt agaaatcatc acaatttata attctgaact ccgcgggatt tgtaattact 4381 acggtctagc aagtaatttt aaccagctca attattttgc ttatcttatg gaatacagct 4441 gtctaaaaac gatagcctcc aaacataagg gaacactttc aaaaaccatt tccatgttta 4501 aagatggaag tggttcgtgg gggatcccgt atgagataaa gcaaggtaag cagcgccgtt 4561 attttgcaaa ttttagtgaa tgtaaatccc cttatcaatt tacggatgag ataagtcaag 4621 ctcctgtatt gtatggctat gcccggaata ctcttgaaaa caggttaaaa gctaaatgtt 4681 gtgaattatg tgggacgtct gatgaaaata cttcctatga aattcaccat gtcaataagg 4741 tcaaaaatct taaaggcaaa gaaaaatggg aaatggcaat gatagcgaaa caacgtaaaa 4801 ctcttgttgt atgctttcat tgtcatcgtc acgtgattca taaacacaag tgagatgggt 4861 accctcgagg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 4921 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga 4981 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgggtacc 5041 gagctcgaat tcggcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 5101 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga 5161 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat 5221 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cggatcggtc ttgccttgct 5281 cgtcggtgat gtacttcacc agctccgcga agtcgctctt cttgatggag cgcatgggga 5341 cgtgcttggc aatcacgcgc accccccggc cgttttagcg gctaaaaaag tcatggctct 5401 gccctcgggc ggaccacgcc catcatgacc ttgccaagct cgtcctgctt ctcttcgatc 5461 ttcgccagca gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg 5521 agccagagtt tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg 5581 cggacgtgct catagtccac gacgcccgtg attttgtagc cctggccgac ggccagcagg 5641 taggccgaca ggctcatgcc ggccgccgcc gccttttcct caatcgctct tcgttcgtct 5701 ggaaggcagt acaccttgat aggtgggctg cccttcctgg ttggcttggt ttcatcagcc 5761 atccgcttgc cctcatctgt tacgccggcg gtagccggcc agcctcgcag agcaggattc 5821 ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt 5881 gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca 5941 cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc 6001 gctataatga ccccgaagca gggttatgca gcggaaaaga tccgtcgacc ttagaagcaa 6061 acttaagagt gtgttgatag tgcagtatct taaaattttg tgtataatag gaattgaagt 6121 taaattagat gctaaaaatt tgtaattaag aaggagggat tcgtcatgtt ggtattccaa 6181 atgcgtaatg tagataaaac atctactgtt ttgaaacaga ctaaaaacag tgattacgca 6241 gataaataaa tacgttagat taattcctac cagtgactaa tcttatgact ttttaaacag 6301 ataactaaaa ttacaaacaa atcgtttaac ttctgtattt atttacagat gtaatcactt 6361 caggagtaat tacatgaaca aaaatataaa atattctcaa aactttttaa cgagtgaaaa 6421 agtactcaac caaataataa aacaattgaa tttaaaagaa accgataccg tttacgaaat 6481 tggaacaggt aaagggcatt taacgacgaa actggctaaa ataagtaaac aggtaacgtc 6541 tattgaatta gacagtcatc tattcaactt atcgtcagaa aaattaaaac tgaacattcg 6601 tgtcacttta attcaccaag atattctaca gtttcaattc cctaacaaac agaggtataa 6661 aattgttggg agtattcctt accatttaag cacacaaatt attaaaaaag tggtttttga 6721 aagccatgcg tctgacatct atctgattgt tgaagaagga ttctacaagc gtaccttgga 6781 tattcaccga acactagggt tgctcttgca cactcaagtc tcgattcagc aattgcttaa 6841 gctgccagcg gaatgctttc atcctaaacc aaaagtaaac agtgtcttaa taaaacttac 6901 ccgccatacc acagatgttc cagataaata ttggaagcta tatacgtact ttgtttcaaa 6961 atgggtcaat cgagaatatc gtcaactgtt tactaaaaat cagtttcatc aagcaatgaa 7021 acacgccaaa gtaaacaatt taagtaccat tacttatgag caagtattgt ctatttttaa 7081 tagttatcta ttatttaacg ggaggaaata attctatgag tcgctttttt aaatttggaa 7141 agttacacgt tactaaaggg aatggagata aattattaga tatactactg acagcttcca 7201 agaagctaaa gaggtcccta gcgcctacgg ggaatttgta aatcaaaaga aaaattctcc 7261 caagaattag aaatgagtag atcaaattat tcacgaatag aatcaggaaa atcagatcca 7321 accataaaaa cactagaaca aattgcaaag ttaactaact caacgctagt agtggattta 7381 atcccaaatg agccaacaga accagaacca gaaacagaat cagaacaagt aacattggat 7441 ttagaaatgg aagaagaaaa aagcaatgac ttcgtgtgaa taatgcacga aatcgttgct 7501 tatttttttt taaaagcggt atactagata taacgaaaca acgaactgaa tagaaacgaa 7561 aaaagagcca tgacacattt ataaaatgtt tgacgacatt ttataaatgc atagcccgat 7621 aagattgcca aaccaacgct tatcagttag tcagatgaac tcttccctcg taagaagtta 7681 tttaattaac tttgtttgaa gacggtatat aaccgtacta tcattatata gggaaatcag 7741 agagttttca agtatctaag ctactgaatt taagaattgt taagcaatca atcggaaatc 7801 gtttgattgc tttttttgta ttcatttata gaaggtggag tttgtatgaa tcatgatgaa 7861 tgtaaaactt atataaaaaa tagtttattg gagataagaa aattagcaaa tatctataca 7921 ctagaaacgt ttaagaaaga gttagaaaag agaaatatct acttagaaac aaaatcagat 7981 aagtattttt cttcggaggg ggaagattat atatataagt taatagaaaa taacaaaata 8041 atttattcga ttagtggaaa aaaattgact tataaaggaa aaaaatcttt ttcaaaacat 8101 gcaatattga aacagttgaa tgaaaaagca aaccaagtta attaaacaac ctattttata 8161 ggatttatag gaaaggagaa cagctgaatg aatatccctt ttgttgtaga aactgtgctt 8221 catgacggct tgttaaagta caaatttaaa aatagtaaaa ttcgctcaat cactaccaag 8281 ccaggtaaaa gcaaaggggc tatttttgcg tatcgctcaa aatcaagcat gattggcggt 8341 cgtggtgttg ttctgacttc cgaggaagcg attcaagaaa atcaagatac atttacacat 8401 tggacaccca acgtttatcg ttatggaacg tatgcagacg aaaaccgttc atacacgaaa 8461 ggacattctg aaaacaattt aagacaaatc aataccttct ttattgattt tgatattcac 8521 acggcaaaag aaactatttc agcaagcgat attttaacaa ccgctattga tttaggtttt 8581 atgcctacta tgattatcaa atctgataaa ggttatcaag catattttgt tttagaaacg 8641 ccagtctatg tgacttcaaa atcagaattt aaatctgtca aagcagccaa aataatttcg 8701 caaaatatcc gagaatattt tggaaagtct ttgccagttg atctaacgtg taatcatttt 8761 ggtattgctc gcataccaag aacggacaat gtagaatttt ttgatcctaa ttaccgttat 8821 tctttcaaag aatggcaaga ttggtctttc aaacaaacag ataataaggg ctttactcgt 8881 tcaagtctaa cggttttaag cggtacagaa ggcaaaaaac aagtagatga accctggttt 8941 aatctcttat tgcacgaaac gaaattttca ggagaaaagg gtttaatagg gcgtaataac 9001 gtcatgttta ccctctcttt agcctacttt agttcaggct attcaatcga aacgtgcgaa 9061 tataatatgt ttgagtttaa taatcgatta gatcaaccct tagaagaaaa agaagtaatc 9121 aaaattgtta gaagtgccta ttcagaaaac tatcaagggg ctaataggga atacattacc 9181 attctttgca aagcttgggt atcaagtgat ttaaccagta aagatttatt tgtccgtcaa 9241 gggtggttta aattcaagaa aaaaagaagc gaacgtcaac gtgttcattt gtcagaatgg 9301 aaagaagatt taatggctta tattagcgaa aaaagcgatg tatacaagcc ttatttagtg 9361 acgaccaaaa aagagattag agaagtgcta ggcattcctg aacggacatt agataaattg 9421 ctgaaggtac tgaaggcgaa tcaggaaatt ttctttaaga ttaaaccagg aagaaatggt 9481 ggcattcaac ttgctagtgt taaatcattg ttgctatcga tcattaaagt aaaaaaagaa 9541 gaaaaagaaa gctatataaa ggcgctgaca aattcttttg acttagagca tacattcatt 9601 caagagactt taaacaagct agcagaacgc cctaaaacgg acacacaact cgatttgttt 9661 agctatgata caggctgaaa ataaaacccg cactatgcca ttacatttat atctatgata 9721 cgtgtttgtt ttttctttgc tgtttagcga atgattagca gaaatataca gagtaagatt 9781 ttaattaatt attaggggga gaaggagaga gtagcccgaa aacttttagt tggcttggac 9841 tgaacgaagt gagggaaagg ctactaaaac gtcgaggggc agtgagagcg aagcgaacac 9901 ttgatttttt aattttctat cttttatagg tcattagagt atacttattt gtcctataaa 9961 ctatttagca gcataataga tttattgaat aggtcattta agttgagcat attagaggag 10021 gaaaatcttg gagaaatatt tgaagaaccc gattacatgg attggattag ttcttgtggt 10081 tacgtggttt ttaactaaaa gtagtgaatt tttgattttt ggtgtgtgtg tcttgttgtt 10141 agtatttgct agtcaaagtg attaaata //