LOCUS       V016566                 5443 bp    DNA     circular SYN 25-MAY-2021
DEFINITION  Exported.
ACCESSION   V016566
VERSION     V016566
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5443)
  AUTHORS   Mordaka PM, Heap JT.
  TITLE     Combinatorial assembly of integration-activated metabolic pathway
            libraries in Clostridium acetobutlicym
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5443)
  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 5443)
  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            "Unpublished"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..5443
                     /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="QVN25285.1"
                     /translation="MNLNIKRMLKVVTLYDAIIAAIVSVILLFAANYKISLIVIIGIFS
                     AIFNFYLSNLTADFVFVKKMGNTSLIFLSSIFRVILVFFIGIILYKIYKYYLIAYLGGY
                     SAHFIALIIYGSLVNKR"
     regulatory      2843..2884
                     /label="TT2"
                     /note="TT2"
                     /regulatory_class="terminator"
     CDS             3263..3703
                     /codon_start=1
                     /transl_table=11
                     /product="RepL"
                     /label="RepL"
                     /note="plasmid replication protein"
                     /protein_id="QVN25286.1"
                     /translation="MKERYGTVYKGSQRLIDEESGEVIEVDKLYRKQTSGNFVKAYIVQ
                     LISMLDMIGGKKLKIVNYILDNVHLSNNTMIATTREIAKATGTSLQTVITTLKILEEGN
                     IIKRKTGVLMLNPELLMRGDDQKQKYLLLEFGNFEQEANEID"
     CDS             3853..4473
                     /gene="catP"
                     /label="Chloramphenicol acetyltransferase"
                     /note="Chloramphenicol acetyltransferase from Clostridium
                     perfringens. Accession#: P26826"
     rep_origin      4653..5241
                     /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 aaataaaaat aagaagcctg catttgcagg cttcttattt
     2881 ttatggcgcg ccgcattcac ttcttttcta tataaatatg agcgaagcga ataagcgtcg
     2941 gaaaagcagc aaaaagtttc ctttttgctg ttggagcatg ggggttcagg gggtgcagta
     3001 tctgacgtca atgccgagcg aaagcgagcc gaagggtagc atttacgtta gataaccccc
     3061 tgatatgctc cgacgcttta tatagaaaag aagattcaac taggtaaaat cttaatatag
     3121 gttgagatga taaggtttat aaggaatttg tttgttctaa tttttcactc attttgttct
     3181 aatttctttt aacaaatgtt cttttttttt tagaacagtt atgatatagt tagaatagtt
     3241 taaaataagg agtgagaaaa agatgaaaga aagatatgga acagtctata aaggctctca
     3301 gaggctcata gacgaagaaa gtggagaagt catagaggta gacaagttat accgtaaaca
     3361 aacgtctggt aacttcgtaa aggcatatat agtgcaatta ataagtatgt tagatatgat
     3421 tggcggaaaa aaacttaaaa tcgttaacta tatcctagat aatgtccact taagtaacaa
     3481 tacaatgata gctacaacaa gagaaatagc aaaagctaca ggaacaagtc tacaaacagt
     3541 aataacaaca cttaaaatct tagaagaagg aaatattata aaaagaaaaa ctggagtatt
     3601 aatgttaaac cctgaactac taatgagagg cgacgaccaa aaacaaaaat acctcttact
     3661 cgaatttggg aactttgagc aagaggcaaa tgaaatagat tgacctccca ataacaccac
     3721 gtagttattg ggaggtcaat ctatgaaatg cgattaaggg ccggccagtg ggcaagttga
     3781 aaaattcaca aaaatgtggt ataatatctt tgttcattag agcgataaac ttgaatttga
     3841 gagggaactt agatggtatt tgaaaaaatt gataaaaata gttggaacag aaaagagtat
     3901 tttgaccact actttgcaag tgtaccttgt acctacagca tgaccgttaa agtggatatc
     3961 acacaaataa aggaaaaggg aatgaaacta tatcctgcaa tgctttatta tattgcaatg
     4021 attgtaaacc gccattcaga gtttaggacg gcaatcaatc aagatggtga attggggata
     4081 tatgatgaga tgataccaag ctatacaata tttcacaatg atactgaaac attttccagc
     4141 ctttggactg agtgtaagtc tgactttaaa tcatttttag cagattatga aagtgatacg
     4201 caacggtatg gaaacaatca tagaatggaa ggaaagccaa atgctccgga aaacattttt
     4261 aatgtatcta tgataccgtg gtcaaccttc gatggcttta atctgaattt gcagaaagga
     4321 tatgattatt tgattcctat ttttactatg gggaaatatt ataaagaaga taacaaaatt
     4381 atacttcctt tggcaattca agttcatcac gcagtatgtg acggatttca catttgccgt
     4441 tttgtaaacg aattgcagga attgataaat agttaacttc aggtttgtct gtaactaaaa
     4501 acaagtattt aagcaaaaac atcgtagaaa tacggtgttt tttgttaccc taagtttaaa
     4561 ctcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4621 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4681 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4741 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     4801 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4861 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4921 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4981 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5041 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5101 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5161 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5221 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5281 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5341 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5401 tcagtgagcg aggaagcgga agagcgccca atacgcaggg ccc
//
