LOCUS       40924_11316        5834 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCM110, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5834)
  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 5834)
  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 5834)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5834)
  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..5834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      3..712
                     /label=oriV
                     /note="incP origin of replication"
     RBS             820..828
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     regulatory      856..1195
                     /note="PmxaF; Methylobacterium extorquens mxa operon
                     promoter"
                     /regulatory_class="promoter"
     CDS             complement(1576..2223)
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     CDS             2329..3525
                     /label=TcR
                     /note="tetracycline efflux protein"
     CDS             complement(3762..4907)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     CDS             complement(5179..5550)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             complement(5434..5550)
                     /codon_start=1
                     /gene="traJ'"
                     /product="TraJ'"
                     /label=traJ'
                     /note="truncated oriT-recognizing protein"
                     /protein_id="AAK73417.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL"
     gene            complement(5434..5550)
                     /gene="traJ'"
                     /label=traJ'
     oriT            complement(5583..5692)
                     /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 atccatatgt aagatttaaa tgcaaccgtt ttttcggaag gaaatgatga cctcgtttcc
      781 accgaattag cttgcatgcc tgcaggtcga ctctagatca cctccttaag cttcccgctt
      841 ggtcgggccg cttcgcgagg gcccgttgac gacaacggtg cgatgggtcc cggccccggt
      901 caagacgatg ccaatacgtt gcgacactac gccttggcac ttttagaatt gccttatcgt
      961 cctgataaga aatgtccgac cagctaaaga catcgcgtcc aatcaaagcc tagaaaatat
     1021 aggcgaaggg acgctaataa gtctttcata agaccgcgca aatctaaaaa tatccttaga
     1081 ttcacgatgc ggcacttcgg atgacttccg agcgagcctg gaacctcaga aaaacgtctg
     1141 agagataccg cggatcctaa gggcgaattc cagcacactg gcggccgtta ctagtggatc
     1201 cgagctcggt accatcgata gatccgttca gggcatcaaa tgcagactcg tttctgacac
     1261 ttgcagactg gatctatctc atctgcgcaa ggcagaacgt gaagacggcc gccctggacc
     1321 tcgcccgcga gcgccaggcg cacgaggccg gcgcgcggac ccgcgccacg gcccacgagc
     1381 ggacgccgca gcaggagcgc cagaaggccg ccagagaggc cgagcgcggc cgtgaggctt
     1441 ggacgctagg gcagggcatg aaaaagcccg tagcgggctg ctacgggcgt ctgacgcggt
     1501 ggaaaggggg aggggatgtt gtctacatgg ctctgctgta gtgagtgggt tgcgctccgg
     1561 cagcggtcct gatcaatcgt caccctttct cggtccttca acgttcctga caacgagcct
     1621 ccttttcgcc aatccatcga caatcaccgc gagtccctgc tcgaacgctg cgtccggacc
     1681 ggcttcgtcg aaggcgtcta tcgcggcccg caacagcggc gagagcggag cctgttcaac
     1741 ggtgccgccg cgctcgccgg catcgctgtc gccggcctgc tcctcaagca cggccccaac
     1801 agtgaagtag ctgattgtca tcagcgcatt gacggcgtcc ccggccgaaa aacccgcctc
     1861 gcagaggaag cgaagctgcg cgtcggccgt ttccatctgc ggtgcgcccg gtcgcgtgcc
     1921 ggcatggatg cgcgcgccat cgcggtaggc gagcagcgcc tgcctgaagc tgcgggcatt
     1981 cccgatcaga aatgagcgcc agtcgtcgtc ggctctcggc accgaatgcg tatgattctc
     2041 cgccagcatg gcttcggcca gtgcgtcgag cagcgcccgc ttgttcctga agtgccagta
     2101 aagcgccggc tgctgaaccc ccaaccgttc cgccagtttg cgtgtcgtca gaccgtctac
     2161 gccgacctcg ttcaacaggt ccagggcggc acggatcact gtattcggct gcaactttgt
     2221 catgattgac actttatcac tgataaacat aatatgtcca ccaacttatc agtgataaag
     2281 aatccgcgcg ttcaatcgga ccagcggagg ctggtccgga ggccagacgt gaaacccaac
     2341 atacccctga tcgtaattct gagcactgtc gcgctcgacg ctgtcggcat cggcctgatt
     2401 atgccggtgc tgccgggcct cctgcgcgat ctggttcact cgaacgacgt caccgcccac
     2461 tatggcattc tgctggcgct gtatgcgttg gtgcaatttg cctgcgcacc tgtgctgggc
     2521 gcgctgtcgg atcgtttcgg gcggcggcca atcttgctcg tctcgctggc cggcgccact
     2581 gtcgactacg ccatcatggc gacagcgcct ttcctttggg ttctctatat cgggcggatc
     2641 gtggccggca tcaccggggc gactggggcg gtagccggcg cttatattgc cgatatcact
     2701 gatggcgatg agcgcgcgcg gcacttcggc ttcatgagcg cctgtttcgg gttcgggatg
     2761 gtcgcgggac ctgtgctcgg tgggctgatg ggcggtttct ccccccacgc tccgttcttc
     2821 gccgcggcag ccttgaacgg cctcaatttc ctgacgggct gtttcctttt gccggagtcg
     2881 cacaaaggcg aacgccggcc gttacgccgg gaggctctca acccgctcgc ttcgttccgg
     2941 tgggcccggg gcatgaccgt cgtcgccgcc ctgatggcgg tcttcttcat catgcaactt
     3001 gtcggacagg tgccggccgc gctttgggtc attttcggcg aggatcgctt tcactgggac
     3061 gcgaccacga tcggcatttc gcttgccgca tttggcattc tgcattcact cgcccaggca
     3121 atgatcaccg gccctgtagc cgcccggctc ggcgaaaggc gggcactcat gctcggaatg
     3181 attgccgacg gcacaggcta catcctgctt gccttcgcga cacggggatg gatggcgttc
     3241 ccgatcatgg tcctgcttgc ttcgggtggc atcggaatgc cggcgctgca agcaatgttg
     3301 tccaggcagg tggatgagga acgtcagggg cagctgcaag gctcactggc ggcgctcacc
     3361 agcctgacct cgatcgtcgg acccctcctc ttcacggcga tctatgcggc ttctataaca
     3421 acgtggaacg ggtgggcatg gattgcaggc gctgccctct acttgctctg cctgccggcg
     3481 ctgcgtcgcg ggctttggag cggcgcaggg caacgagccg atcgctgatc gtggaaacga
     3541 taggcctatg ccatgcgggt caaggcgact tccggcaagc tatacgcgcc ctagaattgt
     3601 caattttaat cctctgttta tcggcagttc gtagagcgcg ccgtgcgtcc cgagcgatac
     3661 tgagcgaagc aagtgcgtcg agcagtgccc gcttgttcct gaaatgccag taaagcgctg
     3721 gctgctgaac ccccagccgg aactgacccc acaaggccct agcgtttgca atgcaccagg
     3781 tcatcattga cccaggcgtg ttccaccagg ccgctgcctc gcaactcttc gcaggcttcg
     3841 ccgacctgct cgcgccactt cttcacgcgg gtggaatccg atccgcacat gaggcggaag
     3901 gtttccagct tgagcgggta cggctcccgg tgcgagctga aatagtcgaa catccgtcgg
     3961 gccgtcggcg acagcttgcg gtacttctcc catatgaatt tcgtgtagtg gtcgccagca
     4021 aacagcacga cgatttcctc gtcgatcagg acctggcaac gggacgtttt cttgccacgg
     4081 tccaggacgc ggaagcggtg cagcagcgac accgattcca ggtgcccaac gcggtcggac
     4141 gtgaagccca tcgccgtcgc ctgtaggcgc gacaggcatt cctcggcctt cgtgtaatac
     4201 cggccattga tcgaccagcc caggtcctgg caaagctcgt agaacgtgaa ggtgatcggc
     4261 tcgccgatag gggtgcgctt cgcgtactcc aacacctgct gccacaccag ttcgtcatcg
     4321 tcggcccgca gctcgacgcc ggtgtaggtg atcttcacgt ccttgttgac gtggaaaatg
     4381 accttgtttt gcagcgcctc gcgcgggatt ttcttgttgc gcgtggtgaa cagggcagag
     4441 cgggccgtgt cgtttggcat cgctcgcatc gtgtccggcc acggcgcaat atcgaacaag
     4501 gaaagctgca tttccttgat ctgctgcttc gtgtgtttca gcaacgcggc ctgcttggcc
     4561 tcgctgacct gttttgccag gtcctcgccg gcggtttttc gcttcttggt cgtcatagtt
     4621 cctcgcgtgt cgatggtcat cgacttcgcc aaacctgccg cctcctgttc gagacgacgc
     4681 gaacgctcca cggcggccga tggcgcgggc agggcagggg gagccagttg cacgctgtcg
     4741 cgctcgatct tggccgtagc ttgctggacc atcgagccga cggactggaa ggtttcgcgg
     4801 ggcgcacgca tgacggtgcg gcttgcgatg gtttcggcat cctcggcgga aaaccccgcg
     4861 tcgatcagtt cttgcctgta tgccttccgg tcaaacgtcc gattcattca ccctccttgc
     4921 gggattgccc cgactcacgc cggggcaatg tgcccttatt cctgatttga cccgcctggt
     4981 gccttggtgt ccagataatc caccttatcg gcaatgaagt cggtcccgta gaccgtctgg
     5041 ccgtccttct cgtacttggt attccgaatc ttgccctgca cgaataccag ctccgcgaag
     5101 tcgctcttct tgatggagcg catggggacg tgcttggcaa tcacgcgcac cccccggccg
     5161 ttttagcggc taaaaaagtc atggctctgc cctcgggcgg accacgccca tcatgacctt
     5221 gccaagctcg tcctgcttct cttcgatctt cgccagcagg gcgaggatcg tggcatcacc
     5281 gaaccgcgcc gtgcgcgggt cgtcggtgag ccagagtttc agcaggccgc ccaggcggcc
     5341 caggtcgcca ttgatgcggg ccagctcgcg gacgtgctca tagtccacga cgcccgtgat
     5401 tttgtagccc tggccgacgg ccagcaggta ggcctacagg ctcatgccgg ccgccgccgc
     5461 cttttcctca atcgctcttc gttcgtctgg aaggcagtac accttgatag gtgggctgcc
     5521 cttcctggtt ggcttggttt catcagccat ccgcttgccc tcatctgtta cgccggcggt
     5581 agccggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg aataagggac
     5641 agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc ccggctgacg
     5701 ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg tatatcgtgc
     5761 gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg gttatgcagc
     5821 ggaaaagatc cgtc
//
