LOCUS       40924_11331        7974 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCM160, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7974)
  AUTHORS   Marx CJ, Lidstrom ME.
  TITLE     Development of improved versatile broad-host-range vectors for use 
            in methylotrophs and other Gram-negative bacteria
  JOURNAL   Microbiology 147 (Pt 8), 2065-2075 (2001)
  PUBMED    11495985
REFERENCE   2  (bases 1 to 7974)
  AUTHORS   Marx CJ, Lidstrom ME.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-DEC-2000) Microbiology, University of Washington, Box 
            357242, Seattle, WA 98195, USA
REFERENCE   3  (bases 1 to 7974)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7974)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Microbiology 147 (Pt 8), 2065-2075 (2001)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-DEC-2000) Microbiology, University of Washington, Box 357242, 
            Seattle, WA 98195, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      3..712
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    1981..2002
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2017..2047
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2055..2071
                     /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)."
     primer_bind     2079..2095
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2118..2435
                     /note="PmxaF; Methylobacterium extorquens mxa operon
                     promoter"
                     /regulatory_class="promoter"
     primer_bind     2446..2462
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(2475..2531)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2532..2548)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2993..3634)
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     CDS             complement(4415..5227)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(5902..7047)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     CDS             complement(7319..7690)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             complement(7574..7690)
                     /codon_start=1
                     /gene="traJ'"
                     /product="TraJ'"
                     /label=traJ'
                     /note="truncated oriT-recognizing protein"
                     /protein_id="AAK73413.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL"
     gene            complement(7574..7690)
                     /gene="traJ'"
                     /label=traJ'
     oriT            complement(7723..7832)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
ORIGIN
        1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
       61 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc
      121 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
      181 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
      241 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
      301 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
      361 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
      421 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
      481 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
      541 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
      601 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc
      661 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agccaagctg
      721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     1261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1741 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1801 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     1861 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1921 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1981 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     2041 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     2101 ttacgccaag ctagcttccc gcttggtcgg gccgcttcgc gagggcccgt tgacgacaac
     2161 ggtgcgatgg gtcccggccc cggtcaagac gatgccaata cgttgcgaca ctacgccttg
     2221 gcacttttag aattgcctta tcgtcctgat aagaaatgtc cgaccagcta aagacatcgc
     2281 gtccaatcaa agcctagaaa atataggcga agggacgcta ataagtcttt cataagaccg
     2341 cgcaaatcta aaaatatcct tagattcacg atgcggcact tcggatgact tccgagcgag
     2401 cctggaacct cagaaaaacg tctgagagat accgcggatc tcacacagga aacagctatg
     2461 accatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg
     2521 agctcgaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2581 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2641 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     2701 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     2761 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     2821 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2881 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     2941 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gcaccctttc
     3001 tcggtccttc aacgttcctg acaacgagcc tccttttcgc caatccatcg acaatcaccg
     3061 cgagtccctg ctcgaacgct gcgtccggac cggcttcgtc gaaggcgtct atcgcggccc
     3121 gcaacagcgg cgagagcgga gcctgttcaa cggtgccgcc gcgctcgccg gcatcgctgt
     3181 cgccggcctg ctcctcaagc acggccccaa cagtgaagta gctgattgtc atcagcgcat
     3241 tgacggcgtc cccggccgaa aaacccgcct cgcagaggaa gcgaagctgc gcgtcggccg
     3301 tttccatctg cggtgcgccc ggtcgcgtgc cggcatggat gcgcgcgcca tcgcggtagg
     3361 cgagcagcgc ctgcctgaag ctgcgggcat tcccgatcag aaatgagcgc cagtcgtcgt
     3421 cggctctcgg caccgaatgc gtatgattct ccgccagcat ggcttcggcc agtgcgtcga
     3481 gcagcgcccg cttgttcctg aagtgccagt aaagcgccgg ctgctgaacc cccaaccgtt
     3541 ccgccagttt gcgtgtcgtc agaccgtcta cgccgacctc gttcaacagg tccagggcgg
     3601 cacggatcac tgtattcggc tgcaactttg tcatgattga cactttatca ctgataaaca
     3661 taatatgtcc accaacttat cagtgataaa gaatccgcgc gttcaatcgg accagcggag
     3721 gctggtccgg aggccagacg tgaaacccaa catacccctg atcgtaattc tgagcactgt
     3781 cgcgctcgac gctgtcggca tcggcctgat tatgccggtg ctgccgggcc tcctgcgcga
     3841 tctggttcac tcgaacgacg tcaccgccca ctatggcatt ctgctggcgc tgtatgcgtt
     3901 ggtgcaattt gcctgcgcac ctgtgctggg cgcgctgtcg gatcgtttcg ggcggcggcc
     3961 aatcttgctc gtctcgctgg ccggcgccac tgtcgactac gccatcatgg cgacagcgcc
     4021 tttcctttgg gttctctata tcgggcggat cgtggccggc atcaccgggg cgactggggc
     4081 ggtagccggc gcttatattg ccgatgacct gcaggggggg gggggcgctg aggtctgcct
     4141 cgtgaagaag gtgttgctga ctcataccag gcctgaatcg ccccatcatc cagccagaaa
     4201 gtgagggagc cacggttgat gagagctttg ttgtaggtgg accagttggt gattttgaac
     4261 ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac
     4321 tcagcaaaag ttcgatttat tcaacaaagc cgccgtcccg tcaagtcagc gtaatgctct
     4381 gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
     4441 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
     4501 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
     4561 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
     4621 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
     4681 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
     4741 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
     4801 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
     4861 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
     4921 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
     4981 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
     5041 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
     5101 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     5161 aatcagcatc catgttggaa tttaatcgcg gcctcgagca agacgtttcc cgttgaatat
     5221 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     5281 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttccc
     5341 ccccccccct gcaggtccga cacggggatg gatggcgttc ccgatcatgg tcctgcttgc
     5401 ttcgggtggc atcggaatgc cggcgctgca agcaatgttg tccaggcagg tggatgagga
     5461 acgtcagggg cagctgcaag gctcactggc ggcgctcacc agcctgacct cgatcgtcgg
     5521 acccctcctc ttcacggcga tctatgcggc ttctataaca acgtggaacg ggtgggcatg
     5581 gattgcaggc gctgccctct acttgctctg cctgccggcg ctgcgtcgcg ggctttggag
     5641 cggcgcaggg caacgagccg atcgctgatc gtggaaacga taggcctatg ccatgcgggt
     5701 caaggcgact tccggcaagc tatacgcgcc ctagaattgt caattttaat cctctgttta
     5761 tcggcagttc gtagagcgcg ccgtgcgtcc cgagcgatac tgagcgaagc aagtgcgtcg
     5821 agcagtgccc gcttgttcct gaaatgccag taaagcgctg gctgctgaac ccccagccgg
     5881 aactgacccc acaaggccct agcgtttgca atgcaccagg tcatcattga cccaggcgtg
     5941 ttccaccagg ccgctgcctc gcaactcttc gcaggcttcg ccgacctgct cgcgccactt
     6001 cttcacgcgg gtggaatccg atccgcacat gaggcggaag gtttccagct tgagcgggta
     6061 cggctcccgg tgcgagctga aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg
     6121 gtacttctcc catatgaatt tcgtgtagtg gtcgccagca aacagcacga cgatttcctc
     6181 gtcgatcagg acctggcaac gggacgtttt cttgccacgg tccaggacgc ggaagcggtg
     6241 cagcagcgac accgattcca ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc
     6301 ctgtaggcgc gacaggcatt cctcggcctt cgtgtaatac cggccattga tcgaccagcc
     6361 caggtcctgg caaagctcgt agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt
     6421 cgcgtactcc aacacctgct gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc
     6481 ggtgtaggtg atcttcacgt ccttgttgac gtggaaaatg accttgtttt gcagcgcctc
     6541 gcgcgggatt ttcttgttgc gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat
     6601 cgctcgcatc gtgtccggcc acggcgcaat atcgaacaag gaaagctgca tttccttgat
     6661 ctgctgcttc gtgtgtttca gcaacgcggc ctgcttggcc tcgctgacct gttttgccag
     6721 gtcctcgccg gcggtttttc gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat
     6781 cgacttcgcc aaacctgccg cctcctgttc gagacgacgc gaacgctcca cggcggccga
     6841 tggcgcgggc agggcagggg gagccagttg cacgctgtcg cgctcgatct tggccgtagc
     6901 ttgctggacc atcgagccga cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg
     6961 gcttgcgatg gtttcggcat cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta
     7021 tgccttccgg tcaaacgtcc gattcattca ccctccttgc gggattgccc cgactcacgc
     7081 cggggcaatg tgcccttatt cctgatttga cccgcctggt gccttggtgt ccagataatc
     7141 caccttatcg gcaatgaagt cggtcccgta gaccgtctgg ccgtccttct cgtacttggt
     7201 attccgaatc ttgccctgca cgaataccag ctccgcgaag tcgctcttct tgatggagcg
     7261 catggggacg tgcttggcaa tcacgcgcac cccccggccg ttttagcggc taaaaaagtc
     7321 atggctctgc cctcgggcgg accacgccca tcatgacctt gccaagctcg tcctgcttct
     7381 cttcgatctt cgccagcagg gcgaggatcg tggcatcacc gaaccgcgcc gtgcgcgggt
     7441 cgtcggtgag ccagagtttc agcaggccgc ccaggcggcc caggtcgcca ttgatgcggg
     7501 ccagctcgcg gacgtgctca tagtccacga cgcccgtgat tttgtagccc tggccgacgg
     7561 ccagcaggta ggcctacagg ctcatgccgg ccgccgccgc cttttcctca atcgctcttc
     7621 gttcgtctgg aaggcagtac accttgatag gtgggctgcc cttcctggtt ggcttggttt
     7681 catcagccat ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag
     7741 caggattccc gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc
     7801 tcgcgggtgg gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa
     7861 agtctacacg aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg
     7921 aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc ggaaaagatc cgtc
//
