LOCUS       40924_11386        7245 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCM80, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7245)
  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 7245)
  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 7245)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7245)
  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..7245
                     /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             3740..4936
                     /label=TcR
                     /note="tetracycline efflux protein"
     CDS             complement(5173..6318)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     CDS             complement(6590..6961)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             complement(6845..6961)
                     /codon_start=1
                     /gene="traJ'"
                     /product="TraJ'"
                     /label=traJ'
                     /note="truncated oriT-recognizing protein"
                     /protein_id="AAK73409.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL"
     gene            complement(6845..6961)
                     /gene="traJ'"
                     /label=traJ'
     oriT            complement(6994..7103)
                     /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 ccgatatcac tgatggcgat gagcgcgcgc ggcacttcgg
     4141 cttcatgagc gcctgtttcg ggttcgggat ggtcgcggga cctgtgctcg gtgggctgat
     4201 gggcggtttc tccccccacg ctccgttctt cgccgcggca gccttgaacg gcctcaattt
     4261 cctgacgggc tgtttccttt tgccggagtc gcacaaaggc gaacgccggc cgttacgccg
     4321 ggaggctctc aacccgctcg cttcgttccg gtgggcccgg ggcatgaccg tcgtcgccgc
     4381 cctgatggcg gtcttcttca tcatgcaact tgtcggacag gtgccggccg cgctttgggt
     4441 cattttcggc gaggatcgct ttcactggga cgcgaccacg atcggcattt cgcttgccgc
     4501 atttggcatt ctgcattcac tcgcccaggc aatgatcacc ggccctgtag ccgcccggct
     4561 cggcgaaagg cgggcactca tgctcggaat gattgccgac ggcacaggct acatcctgct
     4621 tgccttcgcg acacggggat ggatggcgtt cccgatcatg gtcctgcttg cttcgggtgg
     4681 catcggaatg ccggcgctgc aagcaatgtt gtccaggcag gtggatgagg aacgtcaggg
     4741 gcagctgcaa ggctcactgg cggcgctcac cagcctgacc tcgatcgtcg gacccctcct
     4801 cttcacggcg atctatgcgg cttctataac aacgtggaac gggtgggcat ggattgcagg
     4861 cgctgccctc tacttgctct gcctgccggc gctgcgtcgc gggctttgga gcggcgcagg
     4921 gcaacgagcc gatcgctgat cgtggaaacg ataggcctat gccatgcggg tcaaggcgac
     4981 ttccggcaag ctatacgcgc cctagaattg tcaattttaa tcctctgttt atcggcagtt
     5041 cgtagagcgc gccgtgcgtc ccgagcgata ctgagcgaag caagtgcgtc gagcagtgcc
     5101 cgcttgttcc tgaaatgcca gtaaagcgct ggctgctgaa cccccagccg gaactgaccc
     5161 cacaaggccc tagcgtttgc aatgcaccag gtcatcattg acccaggcgt gttccaccag
     5221 gccgctgcct cgcaactctt cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg
     5281 ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc ttgagcgggt acggctcccg
     5341 gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc gacagcttgc ggtacttctc
     5401 ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg acgatttcct cgtcgatcag
     5461 gacctggcaa cgggacgttt tcttgccacg gtccaggacg cggaagcggt gcagcagcga
     5521 caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg
     5581 cgacaggcat tcctcggcct tcgtgtaata ccggccattg atcgaccagc ccaggtcctg
     5641 gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc
     5701 caacacctgc tgccacacca gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt
     5761 gatcttcacg tccttgttga cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat
     5821 tttcttgttg cgcgtggtga acagggcaga gcgggccgtg tcgtttggca tcgctcgcat
     5881 cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc atttccttga tctgctgctt
     5941 cgtgtgtttc agcaacgcgg cctgcttggc ctcgctgacc tgttttgcca ggtcctcgcc
     6001 ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc
     6061 caaacctgcc gcctcctgtt cgagacgacg cgaacgctcc acggcggccg atggcgcggg
     6121 cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc ttggccgtag cttgctggac
     6181 catcgagccg acggactgga aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat
     6241 ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg
     6301 gtcaaacgtc cgattcattc accctccttg cgggattgcc ccgactcacg ccggggcaat
     6361 gtgcccttat tcctgatttg acccgcctgg tgccttggtg tccagataat ccaccttatc
     6421 ggcaatgaag tcggtcccgt agaccgtctg gccgtccttc tcgtacttgg tattccgaat
     6481 cttgccctgc acgaatacca gctccgcgaa gtcgctcttc ttgatggagc gcatggggac
     6541 gtgcttggca atcacgcgca ccccccggcc gttttagcgg ctaaaaaagt catggctctg
     6601 ccctcgggcg gaccacgccc atcatgacct tgccaagctc gtcctgcttc tcttcgatct
     6661 tcgccagcag ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga
     6721 gccagagttt cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc
     6781 ggacgtgctc atagtccacg acgcccgtga ttttgtagcc ctggccgacg gccagcaggt
     6841 aggcctacag gctcatgccg gccgccgccg ccttttcctc aatcgctctt cgttcgtctg
     6901 gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
     6961 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
     7021 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     7081 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     7141 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     7201 ctataatgac cccgaagcag ggttatgcag cggaaaagat ccgtc
//
