LOCUS       sequence-12110-e        6972 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 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 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: pQexp
FEATURES             Location/Qualifiers
     source          1..6972
                     /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     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
//
