LOCUS       62056_12165        5238 bp DNA     circular SYN 14-DEC-2020
DEFINITION  Cloning vector pHal7, complete sequence.
ACCESSION   MW076822
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5238)
  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 5238)
  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 5238)
  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..5238
                     /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"
     CDS             1972..2016
                     /label=S-Tag
                     /note="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
     terminator      2125..2219
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2328..3140
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     promoter        3594..3696
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             3916..4704
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     oriT            4885..4993
                     /label=oriT
                     /note="incP origin of transfer"
     rep_origin      5087..5238
                     /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 aaatactagc tcgagtctgg taaagaaacc
     1981 gctgctgcga aatttgaacg ccagcacatg gactcgtcta ctagcgcagc ttaattaacc
     2041 taggctgctg ccaccgctga gcaataaagc ccatgcaagc ttgcggccgc gtcgtgactg
     2101 ggaaaaccct ggcgactagt cttggactcc tgttgataga tccagtaatg acctcagaac
     2161 tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt ggtgagaatc
     2221 cagacgttgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     2281 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     2341 aacgggaaac gtcttgctcg aggccgcgat taaattccaa catggatgct gatttatatg
     2401 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg
     2461 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     2521 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     2581 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccgggaaaa
     2641 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     2701 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
     2761 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
     2821 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataagc
     2881 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     2941 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     3001 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     3061 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
     3121 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac tggcagagca
     3181 ttacgctgac ttgacgggac ggcggctttg ttgaataaat cgaacttttg ctgagttgaa
     3241 ggatcagatc acgcatcttc ccgacaacgc agaccgttcc gtggcaaagc aaaagttcaa
     3301 aatcaccaac tggtccacct acaacaaagc tctcatcaac cgtggctccc tcactttctg
     3361 gctggatgat ggggcgattc aggcctggta tgagtcagca acaccttctt cacgaggcag
     3421 acctcagcgc tattctgacc ttgccatcac gactgtgctg gtcattaaac gcgtattcag
     3481 gctgaccctg cgcgctgcgc agggctttat tgattccatt tttacactga tgaatgttcc
     3541 gttgcgctgc ccggattaca gccggatcct ctagagtcga cctgcaggca tgctgatcgg
     3601 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     3661 gagttatcga gattttcagg agctaaggaa gctaaaatgc gctcacgcaa ctggtccaga
     3721 accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac
     3781 tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg ttacgccgtg
     3841 ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag tcgccctaaa
     3901 acaaagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca actatcagag
     3961 gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca tttgtacggc
     4021 tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt tacggtgacc
     4081 gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga aacttcggct
     4141 tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt gcacgacgac
     4201 atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg gcagcgcaat
     4261 gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc tatcttgctg
     4321 acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga actctttgat
     4381 ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct atggaactcg
     4441 ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg catttggtac
     4501 agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc aatggagcgc
     4561 ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct tggacaagaa
     4621 gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta cgtgaaaggc
     4681 gagatcacca aggtagtcgg caaataaact agtaaataat aaaaaagccg gattaataat
     4741 ctggcttttt atattctctg cataaccctg cttcggggtc attatagcga ttttttcggt
     4801 atatccatcc tttttcgcac gatatacagg attttgccaa agggttcgtg tagactttcc
     4861 ttggtgtatc caacggcgtc agccgggcag gataggtgaa gtaggcccac ccgcgagcgg
     4921 gtgttccttc ttcactgtcc cttattcgca cctggcggtg ctcaacggga atcctgctct
     4981 gcgaggctgg ccgtaggccg gccgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5041 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5101 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5161 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5221 taccaaatac tgttcttc
//
