LOCUS       62056_12150        6948 bp DNA     circular SYN 14-DEC-2020
DEFINITION  Cloning vector pHal7-CvFAPG462V, complete sequence.
ACCESSION   MW076827
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6948)
  AUTHORS   Trisrivirat D, Hughes JMX., Hoeven R, Faulkner M, Toogood H, Chaiyen
            P, Scrutton NS.
  TITLE     Promoter engineering for microbial bio-alkane gas production
  JOURNAL   Synth Biol (Oxf) 5 (1), ysaa022 (2020)
  PUBMED    33263086
REFERENCE   2  (bases 1 to 6948)
  AUTHORS   Trisrivirat D, Hughes JMX., Hoeven R, Faulkner M, Toogood HS, 
            Chaiyen P, Scrutton NS.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-OCT-2020) Chemistry, University of Manchester, 131 
            Princess Street, Manchester, Lancashire M1 7DN, United Kingdom
REFERENCE   3  (bases 1 to 6948)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Synth Biol 
            (Oxf)"; date: "2020"; volume: "5"; issue: "1"; pages: "ysaa022"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-OCT-2020) Chemistry, University of Manchester, 131 Princess 
            Street, Manchester, Lancashire M1 7DN, United Kingdom"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(494..1324)
                     /label=pRO1600 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pRO1600 from Pseudomonas aeruginosa"
     rep_origin      1338..1689
                     /label=pRO1600 oriV
                     /note="broad-host-range origin of replication from
                     Pseudomonas aeruginosa plasmid pRO1600; requires the 
                     pRO1600 Rep protein for replication (West et al., 1994)"
     terminator      complement(1762..1848)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     regulatory      1897..1959
                     /regulatory_class="promoter"
     CDS             1960..3744
                     /codon_start=1
                     /transl_table=11
                     /product="CvFAPG462V"
                     /label=CvFAPG462V
                     /protein_id="QPO14559.1"
                     /translation="MASAVEDIRKVLSDSSSPVAGQKYDYILVGGGTAACVLANRLSAD
                     GSKRVLVLEAGPDNTSRDVKIPAAITRLFRSPLDWNLFSELQEQLAERQIYMARGRLLG
                     GSSATNATLYHRGAAGDYDAWGVEGWSSEDVLSWFVQAETNADFGPGAYHGSGGPMRVE
                     NPRYTNKQLHTAFFKAAEEVGLTPNSDFNDWSHDHAGYGTFQVMQDKGTRADMYRQYLK
                     PVLGRRNLQVLTGAAVTKVNIDQAAGKAQALGVEFSTDGPTGERLSAELAPGGEVIMCA
                     GAVHTPFLLKHSGVGPSAELKEFGIPVVSNLAGVGQNLQDQPACLTAAPVKEKYDGIAI
                     SDHIYNEKGQIRKRAIASYLLGGRGGLTSTGCDRGAFVRTAGQALPDLQVRFVPGMALD
                     PDVVSTYVRFAKFQSQGLKWPSGITMQLIACRPQSTGSVGLKSADPFAPPKLSPGYLTD
                     KDGADLATLRKGIHWARDVARSSALSEYLDGELFPGSGVVSDDQIDEYIRRSIHSSNAI
                     TGTCKMGNAGDSSSVVDNQLRVHGVEGLRVVDASVVPKIPGGQTGAPVVMIAERAAALL
                     TGKATIGASAAAPATVAA"
     terminator      3835..3929
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4038..4850
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        5304..5406
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             5626..6414
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     oriT            6595..6703
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      6797..6948
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
       61 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
      121 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
      181 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
      241 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggcatccg gtaagcggca
      301 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
      361 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
      421 ggcggagcct atggaaaaac gccagcaacg cggccgtgaa aggcaggccg gtccgtggtg
      481 gccacggcct ctaggccaga tccagcggca tctgggttag tcgagcgcgg gccgcttccc
      541 atgtctcacc agggcgagcc tgtttcgcga tctcagcatc tgaaatcttc ccggccttgc
      601 gcttcgctgg ggccttaccc accgccttgg cgggcttctt cggtccaaaa ctgaacaaca
      661 gatgtgtgac cttgcgcccg gtctttcgct gcgcccactc cacctgtagc gggctgtgct
      721 cgttgatctg cgtcacggct ggatcaagca ctcgcaactt gaagtccttg atcgagggat
      781 accggccttc cagttgaaac cactttcgca gctggtcaat ttctatttcg cgctggccga
      841 tgctgtccca ttgcatgagc agctcgtaaa gcctgatcgc gtgggtgctg tccatcttgg
      901 ccacgtcagc caaggcgtat ttggtgaact gtttggtgag ttccgtcagg tacggcagca
      961 tgtctttggt gaacctgagt tctacacggc cctcaccctc ccggtagatg attgtttgca
     1021 cccagccggt aatcatcaca ctcggtcttt tccccttgcc attgggctct tgggttaacc
     1081 ggacttcccg ccgtttcagg cgcagggccg cttctttgag ctggttgtag gaagattcga
     1141 tagggacacc cgccatcgtc gctatgtcct ccgccgtcac tgaatacatc acttcatcgg
     1201 tgacaggctc gctcctcttc acctggctaa tacaggccag aacgatccgc tgttcctgaa
     1261 cactgaggcg atacgcggcc tcgaccaggg cattgctttt gtaaaccatt gggggtgagg
     1321 ccacgttcga cattccttgt gtataagggg acactgtatc tgcgtcccac aatacaacaa
     1381 atccgtccct ttacaacaac aaatccgtcc cttcttaaca acaaatccgt cccttaatgg
     1441 caacaaatcc gtcccttttt aaactctaga ggccacggat tacgtggcct gtagacgtcc
     1501 taaaaggttt aaaagggaaa aggaagaaaa gggtggaaac gcaaaaaacg caccactacg
     1561 tggccccgtt ggggccgcat ttgtgcccct gaaggggcgg gggaggcgtc tgggcaatcc
     1621 ccgttttacc agtcccctat cgccgcctga gagggcgcag gaagcgagta atcagggtat
     1681 cgaggcggat tcacccttgg cgtccaacca gcggcaccag cggcgcctga gaggggcgcg
     1741 cccagctgtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     1801 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctttaatt
     1861 aaagcggata acaatttcac acaggaggtt tttctattgc gtacaaccga taaaggtata
     1921 gagtttgaga ctttactaga gaaagaggag aaatactaga tggccagcgc agttgaagat
     1981 attcgtaaag ttctgagcga tagcagcagt ccggttgcag gtcagaaata tgattatatt
     2041 ctggttggtg gtggcaccgc agcatgtgtt ctggcaaatc gtctgagcgc agatggtagc
     2101 aaacgtgttc tggttctgga agcaggtccg gataatacca gccgtgatgt taaaattccg
     2161 gcagcaatta cccgtctgtt tcgtagtccg ctggattgga acctgtttag cgaactgcaa
     2221 gaacagctgg cagaacgtca gatttatatg gcacgtggtc gtctgttagg tggtagcagc
     2281 gcaaccaatg caaccctgta tcatcgtggt gcagccggtg attatgatgc atggggtgtt
     2341 gaaggttgga gcagcgaaga tgttctgagc tggtttgttc aggcagaaac caatgccgat
     2401 tttggtccgg gtgcatatca tggtagcggt ggtccgatgc gtgttgaaaa tccgcgttat
     2461 accaataaac agctgcatac cgcatttttc aaagcagcag aagaagttgg tctgaccccg
     2521 aatagcgatt ttaatgattg gagccatgat cacgcaggtt atggcacctt tcaggttatg
     2581 caggataaag gcacccgtgc agatatgtat cgtcagtatc tgaaaccggt tctgggtcgt
     2641 cgtaatctgc aggttctgac cggtgcagca gttaccaaag ttaatattga tcaggcagca
     2701 ggtaaagcac aggcactggg cgttgaattt tcaaccgatg gtccgaccgg tgaacgtctg
     2761 tcagcagaac tggcacctgg tggtgaagtt attatgtgtg ccggtgcagt tcataccccg
     2821 tttctgctga aacatagcgg tgttggtccg agcgcagaac tgaaagaatt tggtattccg
     2881 gttgttagca atctggcagg cgttggccag aatctgcaag atcagcctgc atgtctgacc
     2941 gcagcaccgg ttaaagaaaa atatgatggt attgccatca gcgaccacat ctataatgaa
     3001 aaaggtcaga ttcgcaaacg tgccattgca agctatctgc ttggtggtcg tggtggcctg
     3061 accagtaccg gttgtgatcg cggtgcattt gttcgtaccg caggtcaggc actgccggat
     3121 ctgcaagttc gttttgttcc tggtatggca ctggatccgg atgttgttag cacctatgtg
     3181 cgttttgcca aatttcagag ccagggcctg aaatggccga gcggtattac catgcagctg
     3241 attgcatgtc gtccgcagag taccggtagc gtgggtctga aaagcgcaga tccgtttgca
     3301 ccgcctaaac tgagtccggg ttatctgacc gataaagatg gtgcagatct ggcgaccctg
     3361 cgtaaaggta ttcattgggc acgtgatgtt gcacgtagct cagcactgag cgaatatctg
     3421 gatggtgaac tgtttcctgg tagtggtgtt gttagtgatg atcagatcga tgaatatatc
     3481 cgtcgtagca ttcatagcag caatgcaatt accggcacct gtaaaatggg taatgccggt
     3541 gatagcagct cagttgttga taaccagctg cgtgtgcatg gtgtggaagg tctgcgtgtt
     3601 gttgatgcaa gcgttgttcc gaaaattcct ggtggtcaga caggtgctcc ggtggtgatg
     3661 attgcagaac gtgcagcagc cctgctgacc ggtaaagcaa ccattggtgc aagcgcagcc
     3721 gcaccggcaa ccgttgcagc ataattaacc taggctgctg ccaccgctga gcaataaagc
     3781 ccatgcaagc ttgcggccgc gtcgtgactg ggaaaaccct ggcgactagt cttggactcc
     3841 tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga acgctcggtt
     3901 gccgccgggc gttttttatt ggtgagaatc cagacgttgt gtctcaaaat ctctgatgtt
     3961 acattgcaca agataaaaat atatcatcat gaacaataaa actgtctgct tacataaaca
     4021 gtaatacaag gggtgttatg agccatattc aacgggaaac gtcttgctcg aggccgcgat
     4081 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     4141 aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga
     4201 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     4261 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     4321 ggttactcac cactgcgatc cccgggaaaa cagcattcca ggtattagaa gaatatcctg
     4381 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     4441 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     4501 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     4561 ttgaacaagt ctggaaagaa atgcataagc ttttgccatt ctcaccggat tcagtcgtca
     4621 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     4681 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     4741 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     4801 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     4861 tggttaattg gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcggctttg
     4921 ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc
     4981 agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct acaacaaagc
     5041 tctcatcaac cgtggctccc tcactttctg gctggatgat ggggcgattc aggcctggta
     5101 tgagtcagca acaccttctt cacgaggcag acctcagcgc tattctgacc ttgccatcac
     5161 gactgtgctg gtcattaaac gcgtattcag gctgaccctg cgcgctgcgc agggctttat
     5221 tgattccatt tttacactga tgaatgttcc gttgcgctgc ccggattaca gccggatcct
     5281 ctagagtcga cctgcaggca tgctgatcgg cacgtaagag gttccaactt tcaccataat
     5341 gaaataagat cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa
     5401 gctaaaatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc
     5461 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg
     5521 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     5581 gatgttacgc agcagggcag tcgccctaaa acaaagttaa acatcatgag ggaagcggtg
     5641 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
     5701 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
     5761 agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
     5821 ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct
     5881 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
     5941 gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc
     6001 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
     6061 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
     6121 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
     6181 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     6241 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     6301 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     6361 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaact
     6421 agtaaataat aaaaaagccg gattaataat ctggcttttt atattctctg cataaccctg
     6481 cttcggggtc attatagcga ttttttcggt atatccatcc tttttcgcac gatatacagg
     6541 attttgccaa agggttcgtg tagactttcc ttggtgtatc caacggcgtc agccgggcag
     6601 gataggtgaa gtaggcccac ccgcgagcgg gtgttccttc ttcactgtcc cttattcgca
     6661 cctggcggtg ctcaacggga atcctgctct gcgaggctgg ccgtaggccg gccgataatc
     6721 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6781 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6841 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6901 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttc
//
