LOCUS       V016385                 5401 bp    DNA     circular SYN 25-MAY-2021
DEFINITION  Exported.
ACCESSION   V016385
VERSION     V016385
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5401)
  AUTHORS   Mordaka PM, Heap JT.
  TITLE     Combinatorial assembly of integration-activated metabolic pathway
            libraries in Clostridium acetobutlicym
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5401)
  AUTHORS   Mordaka PM, Heap JT.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-APR-2020) Life Sciences, Imperial College London,
            Imperial College Road, London SW7 2AY, United Kingdom
REFERENCE   3  (bases 1 to 5401)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (17-APR-2020) Life Sciences, Imperial College London, Imperial
            College Road, London SW7 2AY, United Kingdom"
FEATURES             Location/Qualifiers
     source          1..5401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      9..62
                     /label="derived from CD0164 gene of Clostridium difficile"
                     /note="derived from CD0164 gene of Clostridium difficile"
                     /regulatory_class="terminator"
     primer_bind     63..79
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    90..389
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium acetobutylicum ATCC824"
     CDS             413..1147
                     /gene="ermBP"
                     /label="rRNA adenine N-6-methyltransferase"
                     /note="rRNA adenine N-6-methyltransferase from Enterococcus
                     faecalis. Accession#: P0A4D5"
     protein_bind    1192..1213
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1228..1258
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    1266..1282
                     /label="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)."
     misc_feature    complement(1319..1375)
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1376..1392)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    1643..2842
                     /note="mediates homologous recombination with the
                     chromosome of Clostridium acetobutylicum ATCC824"
     CDS             2078..2443
                     /codon_start=1
                     /transl_table=11
                     /product="CAC2872"
                     /label="CAC2872"
                     /note="putative FoF1-type ATP synthase operon membrane
                     protein"
                     /protein_id="QVN25280.1"
                     /translation="MNLNIKRMLKVVTLYDAIIAAIVSVILLFAANYKISLIVIIGIFS
                     AIFNFYLSNLTADFVFVKKMGNTSLIFLSSIFRVILVFFIGIILYKIYKYYLIAYLGGY
                     SAHFIALIIYGSLVNKR"
     CDS             3221..3661
                     /codon_start=1
                     /transl_table=11
                     /product="RepL"
                     /label="RepL"
                     /note="plasmid replication protein"
                     /protein_id="QVN25281.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     CDS             3811..4431
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      4611..5199
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 cctgcaggat aaaaaaattg tagataaatt ttataaaata gttttatcta caattttttt
       61 atcaggaaac agctatgacc gcggccatct atgcaacaaa agcagctatt gaaaaagcag
      121 gttggacagt tgatgaatta gatttaatag aatcaaatga agcttttgca gctcaaagtt
      181 tagcagtagc aaaagattta aaatttgata tgaataaagt aaatgtaaat ggaggagcta
      241 ttgcccttgg tcatccaatt ggagcatcag gtgcaagaat actcgttact cttgtacacg
      301 caatgcaaaa aagagatgca aaaaaaggct tagcaacttt atgtataggt ggcggacaag
      361 gaacagcaat attgctagaa aagtgctaga tcgattaaga aggagtgatt acatgaacaa
      421 aaatataaaa tattctcaaa actttttaac gagtgaaaaa gtactcaacc aaataataaa
      481 acaattgaat ttaaaagaaa ccgataccgt ttacgaaatt ggaacaggta aagggcattt
      541 aacgacgaaa ctggctaaaa taagtaaaca ggtaacgtct attgaattag acagtcatct
      601 attcaactta tcgtcagaaa aattaaaact gaatactcgt gtcactttaa ttcaccaaga
      661 tattctacag tttcaattcc ctaacaaaca gaggtataaa attgttggga gtattcctta
      721 ccatttaagc acacaaatta ttaaaaaagt ggtttttgaa agccatgcgt ctgacatcta
      781 tctgattgtt gaagaaggat tctacaagcg taccttggat attcaccgaa cactagggtt
      841 gctcttgcac actcaagtct cgattcagca attgcttaag ctgccagcgg aatgctttca
      901 tcctaaacca aaagtaaaca gtgtcttaat aaaacttacc cgccatacca cagatgttcc
      961 agataaatat tggaagctat atacgtactt tgtttcaaaa tgggtcaatc gagaatatcg
     1021 tcaactgttt actaaaaatc agtttcatca agcaatgaaa cacgccaaag taaacaattt
     1081 aagtaccgtt acttatgagc aagtattgtc tatttttaat agttatctat tatttaacgg
     1141 gaggaaataa agcggccgcg gagagagacc gggcagtgag cgcaacgcaa ttaatgtgag
     1201 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
     1261 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
     1321 gcttgcatgc ctgcaggtcg actctagagg atccccgggt accgagctcg aattcactgg
     1381 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     1441 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     1501 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
     1561 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     1621 catagggtct catactgcta gcaaagtatt gttaaaaata actctgtaga attataaatt
     1681 agttctacag agttattttt taaaaaaatt ctaaacttat gtataaaaaa tacgataaga
     1741 atgtagaatt aaaactaaag acagttcaat ttcttttaga ataatttagt tagtgtggta
     1801 aaaaaatgtc ataatgatat ttatgttgaa atttgtataa aattcagaaa atgaatatat
     1861 tttatcaatt ttcagtcatt tgaaagatta tgaggctaat gcagtactag gcgtaaattg
     1921 aatttataat tactatagcg ataagaaatg gcctaaaaac gtttgcagta atgaaagaac
     1981 cgtaaatatt ataaaaaaaa tcttaaaaca gagttttatt tataaaaatt taagatatat
     2041 aatttaaata acgtgttaaa atagtggagg aagtaatttg aatctgaata ttaaaagaat
     2101 gttaaaggtt gtaactcttt atgatgcaat tattgctgca atagtttcag taatactttt
     2161 gtttgctgct aattataaga tttcgttaat agtgattata gggatttttt cagcaatatt
     2221 taatttttat ttaagtaatt taacagctga tttcgttttt gtaaaaaaaa tgggaaatac
     2281 gtcacttata tttcttagtt caatttttag agtaatactt gtttttttta taggtattat
     2341 tctttataaa atatataaat attatttaat agcctactta ggaggatata gtgctcattt
     2401 tatagccctt ataatttatg ggtcactagt aaataaacga tgaaaggaag tgattgaatg
     2461 gagctaggtg caaagacagt attttcgatg aagcttggaa gttacaactt tgctataaca
     2521 gaaactgtag tattacagtg gattatcatg gcagttataa tattacttgc aatatttctt
     2581 actaaaaatc ttaagaaagt accaaatagg aaacaaagcg taatagaaat gattgttaac
     2641 ttaataaatg gattggtaaa agaaaatatg ggagagaaat tcatgaattt cgttccaatt
     2701 atcggtacta tggcagtgtt tatacttttc ttaaatttaa cagggctagt aggtatcgaa
     2761 ccagcaacaa aggatattag tgttacagca ggctttgctt tagtaagtgc atttttaata
     2821 aatgcaactg caataaaaag aaggcgcgcc gcattcactt cttttctata taaatatgag
     2881 cgaagcgaat aagcgtcgga aaagcagcaa aaagtttcct ttttgctgtt ggagcatggg
     2941 ggttcagggg gtgcagtatc tgacgtcaat gccgagcgaa agcgagccga agggtagcat
     3001 ttacgttaga taaccccctg atatgctccg acgctttata tagaaaagaa gattcaacta
     3061 ggtaaaatct taatataggt tgagatgata aggtttataa ggaatttgtt tgttctaatt
     3121 tttcactcat tttgttctaa tttcttttaa caaatgttct ttttttttta gaacagttat
     3181 gatatagtta gaatagttta aaataaggag tgagaaaaag atgaaagaaa gatatggaac
     3241 agtctataaa ggctctcaga ggctcataga cgaagaaagt ggagaagtca tagaggtaga
     3301 caagttatac cgtaaacaaa cgtctggtaa cttcgtaaag gcatatatag tgcaattaat
     3361 aagtatgtta gatatgattg gcggaaaaaa acttaaaatc gttaactata tcctagataa
     3421 tgtccactta agtaacaata caatgatagc tacaacaaga gaaatagcaa aagctacagg
     3481 aacaagtcta caaacagtaa taacaacact taaaatctta gaagaaggaa atattataaa
     3541 aagaaaaact ggagtattaa tgttaaaccc tgaactacta atgagaggcg acgaccaaaa
     3601 acaaaaatac ctcttactcg aatttgggaa ctttgagcaa gaggcaaatg aaatagattg
     3661 acctcccaat aacaccacgt agttattggg aggtcaatct atgaaatgcg attaagggcc
     3721 ggccagtggg caagttgaaa aattcacaaa aatgtggtat aatatctttg ttcattagag
     3781 cgataaactt gaatttgaga gggaacttag atggtatttg aaaaaattga taaaaatagt
     3841 tggaacagaa aagagtattt tgaccactac tttgcaagtg taccttgtac ctacagcatg
     3901 accgttaaag tggatatcac acaaataaag gaaaagggaa tgaaactata tcctgcaatg
     3961 ctttattata ttgcaatgat tgtaaaccgc cattcagagt ttaggacggc aatcaatcaa
     4021 gatggtgaat tggggatata tgatgagatg ataccaagct atacaatatt tcacaatgat
     4081 actgaaacat tttccagcct ttggactgag tgtaagtctg actttaaatc atttttagca
     4141 gattatgaaa gtgatacgca acggtatgga aacaatcata gaatggaagg aaagccaaat
     4201 gctccggaaa acatttttaa tgtatctatg ataccgtggt caaccttcga tggctttaat
     4261 ctgaatttgc agaaaggata tgattatttg attcctattt ttactatggg gaaatattat
     4321 aaagaagata acaaaattat acttcctttg gcaattcaag ttcatcacgc agtatgtgac
     4381 ggatttcaca tttgccgttt tgtaaacgaa ttgcaggaat tgataaatag ttaacttcag
     4441 gtttgtctgt aactaaaaac aagtatttaa gcaaaaacat cgtagaaata cggtgttttt
     4501 tgttacccta agtttaaact cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4561 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4621 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4681 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4741 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     4801 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4861 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4921 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4981 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5041 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5101 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5161 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5221 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5281 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     5341 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcagggcc
     5401 c
//
