LOCUS       sequence-12109-e       10168 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol 
            Microbiol. 2009 Dec . 74(6):1300-13."
COMMENT     Sequence Label: pQint
FEATURES             Location/Qualifiers
     source          1..10168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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        join(1299..1304,1305..1322,1323..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
//
