LOCUS       62056_12180        7659 bp DNA     circular SYN 14-DEC-2020
DEFINITION  Cloning vector pHalV-FAPG462VRFP, complete sequence.
ACCESSION   MW076830
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7659)
  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 7659)
  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 7659)
  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..7659
                     /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      1889..1959
                     /regulatory_class="promoter"
     gap             1903..1915
                     /estimated_length=13
     CDS             3778..4452
                     /label=mRFP1
                     /note="monomeric derivative of DsRed (Campbell et al.,
                     2002)"
     terminator      4546..4640
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             4749..5561
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        6015..6117
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             6337..7125
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     oriT            7306..7414
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      7508..7659
                     /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 gtnnnnnnnn nnnnngtata
     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 aggttctgcg ggttctgcgg ccggttctgg cgaatttatg
     3781 gcgagtagcg aagacgttat caaagagttc atgcgtttca aagttcgtat ggaaggttcc
     3841 gttaacggtc acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta cgaaggtacc
     3901 cagaccgcta aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg ggacatcctg
     3961 tccccgcagt tccagtacgg ttccaaagct tacgttaaac acccggctga catcccggac
     4021 tacctgaaac tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa cttcgaagac
     4081 ggtggtgttg ttaccgttac ccaggactcc tccctgcaag acggtgagtt catctacaaa
     4141 gttaaactgc gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa aaaaaccatg
     4201 ggttgggaag cttccaccga acgtatgtac ccggaagacg gtgctctgaa aggtgaaatc
     4261 aaaatgcgtc tgaaactgaa agacggtggt cactacgacg ctgaagttaa aaccacctac
     4321 atggctaaaa aaccggttca gctgccgggt gcttacaaaa ccgacatcaa actggacatc
     4381 acctcccaca acgaagacta caccatcgtt gaacagtacg aacgtgctga aggtcgtcac
     4441 tccaccggtg cttaattaac ctaggctgct gccaccgctg agcaataaag cccatgcaag
     4501 cttgcggccg cgtcgtgact gggaaaaccc tggcgactag tcttggactc ctgttgatag
     4561 atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg
     4621 cgttttttat tggtgagaat ccagacgttg tgtctcaaaa tctctgatgt tacattgcac
     4681 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     4741 ggggtgttat gagccatatt caacgggaaa cgtcttgctc gaggccgcga ttaaattcca
     4801 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     4861 cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     4921 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     4981 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     5041 ccactgcgat ccccgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     5101 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     5161 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     5221 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     5281 tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     5341 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     5401 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     5461 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     5521 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     5581 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcggcttt gttgaataaa
     5641 tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc
     5701 cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa
     5761 ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcctggt atgagtcagc
     5821 aacaccttct tcacgaggca gacctcagcg ctattctgac cttgccatca cgactgtgct
     5881 ggtcattaaa cgcgtattca ggctgaccct gcgcgctgcg cagggcttta ttgattccat
     5941 ttttacactg atgaatgttc cgttgcgctg cccggattac agccggatcc tctagagtcg
     6001 acctgcaggc atgctgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga
     6061 tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg
     6121 cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg
     6181 gttttcatgg cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag
     6241 cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg
     6301 cagcagggca gtcgccctaa aacaaagtta aacatcatga gggaagcggt gatcgccgaa
     6361 gtatcgactc aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg
     6421 ctggccgtac atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt
     6481 gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac
     6541 gaccttttgg aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc
     6601 accattgttg tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa
     6661 tttggagaat ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac
     6721 attgatctgg ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca
     6781 gcggcggagg aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa
     6841 accttaacgc tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt
     6901 acgttgtccc gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct
     6961 gccgactggg caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga
     7021 caggcttatc ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa
     7081 tttgtccact acgtgaaagg cgagatcacc aaggtagtcg gcaaataaac tagtaaataa
     7141 taaaaaagcc ggattaataa tctggctttt tatattctct gcataaccct gcttcggggt
     7201 cattatagcg attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca
     7261 aagggttcgt gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga
     7321 agtaggccca cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt
     7381 gctcaacggg aatcctgctc tgcgaggctg gccgtaggcc ggccgataat ctcatgacca
     7441 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7501 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7561 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7621 ctggcttcag cagagcgcag ataccaaata ctgttcttc
//
