LOCUS       40924_11371        5329 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCM51, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5329)
  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 5329)
  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 5329)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5329)
  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..5329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      3..712
                     /label=oriV
                     /note="incP origin of replication"
     CDS             complement(1071..1718)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             1824..3020
                     /codon_start=1
                     /label=TcR
                     /note="tetracycline efflux protein"
                     /translation="VKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     CDS             complement(3257..4402)
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(4674..5045)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             complement(4929..5045)
                     /codon_start=1
                     /gene="traJ'"
                     /product="TraJ'"
                     /label=traJ'
                     /note="truncated oriT-recognizing protein due to a G to T 
                     transversion at nucleotide 7428 of pDN19X that changes a 
                     TCG codon to TAG. This premature stop codon produces a 
                     traJ' allele that lacks the final 85 of its 123 amino 
                     acids"
                     /protein_id="AAK73395.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL"
     gene            complement(4929..5045)
                     /gene="traJ'"
                     /label=traJ'
     oriT            complement(5078..5187)
                     /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 agccaagctc
      721 gttcagggca tcaaatgcag actcgtttct gacacttgca gactggatct atctcatctg
      781 cgcaaggcag aacgtgaaga cggccgccct ggacctcgcc cgcgagcgcc aggcgcacga
      841 ggccggcgcg cggacccgcg ccacggccca cgagcggacg ccgcagcagg agcgccagaa
      901 ggccgccaga gaggccgagc gcggccgtga ggcttggacg ctagggcagg gcatgaaaaa
      961 gcccgtagcg ggctgctacg ggcgtctgac gcggtggaaa gggggagggg atgttgtcta
     1021 catggctctg ctgtagtgag tgggttgcgc tccggcagcg gtcctgatca atcgtcaccc
     1081 tttctcggtc cttcaacgtt cctgacaacg agcctccttt tcgccaatcc atcgacaatc
     1141 accgcgagtc cctgctcgaa cgctgcgtcc ggaccggctt cgtcgaaggc gtctatcgcg
     1201 gcccgcaaca gcggcgagag cggagcctgt tcaacggtgc cgccgcgctc gccggcatcg
     1261 ctgtcgccgg cctgctcctc aagcacggcc ccaacagtga agtagctgat tgtcatcagc
     1321 gcattgacgg cgtccccggc cgaaaaaccc gcctcgcaga ggaagcgaag ctgcgcgtcg
     1381 gccgtttcca tctgcggtgc gcccggtcgc gtgccggcat ggatgcgcgc gccatcgcgg
     1441 taggcgagca gcgcctgcct gaagctgcgg gcattcccga tcagaaatga gcgccagtcg
     1501 tcgtcggctc tcggcaccga atgcgtatga ttctccgcca gcatggcttc ggccagtgcg
     1561 tcgagcagcg cccgcttgtt cctgaagtgc cagtaaagcg ccggctgctg aacccccaac
     1621 cgttccgcca gtttgcgtgt cgtcagaccg tctacgccga cctcgttcaa caggtccagg
     1681 gcggcacgga tcactgtatt cggctgcaac tttgtcatga ttgacacttt atcactgata
     1741 aacataatat gtccaccaac ttatcagtga taaagaatcc gcgcgttcaa tcggaccagc
     1801 ggaggctggt ccggaggcca gacgtgaaac ccaacatacc cctgatcgta attctgagca
     1861 ctgtcgcgct cgacgctgtc ggcatcggcc tgattatgcc ggtgctgccg ggcctcctgc
     1921 gcgatctggt tcactcgaac gacgtcaccg cccactatgg cattctgctg gcgctgtatg
     1981 cgttggtgca atttgcctgc gcacctgtgc tgggcgcgct gtcggatcgt ttcgggcggc
     2041 ggccaatctt gctcgtctcg ctggccggcg ccactgtcga ctacgccatc atggcgacag
     2101 cgcctttcct ttgggttctc tatatcgggc ggatcgtggc cggcatcacc ggggcgactg
     2161 gggcggtagc cggcgcttat attgccgata tcactgatgg cgatgagcgc gcgcggcact
     2221 tcggcttcat gagcgcctgt ttcgggttcg ggatggtcgc gggacctgtg ctcggtgggc
     2281 tgatgggcgg tttctccccc cacgctccgt tcttcgccgc ggcagccttg aacggcctca
     2341 atttcctgac gggctgtttc cttttgccgg agtcgcacaa aggcgaacgc cggccgttac
     2401 gccgggaggc tctcaacccg ctcgcttcgt tccggtgggc ccggggcatg accgtcgtcg
     2461 ccgccctgat ggcggtcttc ttcatcatgc aacttgtcgg acaggtgccg gccgcgcttt
     2521 gggtcatttt cggcgaggat cgctttcact gggacgcgac cacgatcggc atttcgcttg
     2581 ccgcatttgg cattctgcat tcactcgccc aggcaatgat caccggccct gtagccgccc
     2641 ggctcggcga aaggcgggca ctcatgctcg gaatgattgc cgacggcaca ggctacatcc
     2701 tgcttgcctt cgcgacacgg ggatggatgg cgttcccgat catggtcctg cttgcttcgg
     2761 gtggcatcgg aatgccggcg ctgcaagcaa tgttgtccag gcaggtggat gaggaacgtc
     2821 aggggcagct gcaaggctca ctggcggcgc tcaccagcct gacctcgatc gtcggacccc
     2881 tcctcttcac ggcgatctat gcggcttcta taacaacgtg gaacgggtgg gcatggattg
     2941 caggcgctgc cctctacttg ctctgcctgc cggcgctgcg tcgcgggctt tggagcggcg
     3001 cagggcaacg agccgatcgc tgatcgtgga aacgataggc ctatgccatg cgggtcaagg
     3061 cgacttccgg caagctatac gcgccctaga attgtcaatt ttaatcctct gtttatcggc
     3121 agttcgtaga gcgcgccgtg cgtcccgagc gatactgagc gaagcaagtg cgtcgagcag
     3181 tgcccgcttg ttcctgaaat gccagtaaag cgctggctgc tgaaccccca gccggaactg
     3241 accccacaag gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca
     3301 ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca
     3361 cgcgggtgga atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct
     3421 cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact
     3481 tctcccatat gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga
     3541 tcaggacctg gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca
     3601 gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta
     3661 ggcgcgacag gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt
     3721 cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt
     3781 actccaacac ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt
     3841 aggtgatctt cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg
     3901 ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc
     3961 gcatcgtgtc cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct
     4021 gcttcgtgtg tttcagcaac gcggcctgct tggcctcgct gacctgtttt gccaggtcct
     4081 cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact
     4141 tcgccaaacc tgccgcctcc tgttcgagac gacgcgaacg ctccacggcg gccgatggcg
     4201 cgggcagggc agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct
     4261 ggaccatcga gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg
     4321 cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct
     4381 tccggtcaaa cgtccgattc attcaccctc cttgcgggat tgccccgact cacgccgggg
     4441 caatgtgccc ttattcctga tttgacccgc ctggtgcctt ggtgtccaga taatccacct
     4501 tatcggcaat gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac ttggtattcc
     4561 gaatcttgcc ctgcacgaat accagctccg cgaagtcgct cttcttgatg gagcgcatgg
     4621 ggacgtgctt ggcaatcacg cgcacccccc ggccgtttta gcggctaaaa aagtcatggc
     4681 tctgccctcg ggcggaccac gcccatcatg accttgccaa gctcgtcctg cttctcttcg
     4741 atcttcgcca gcagggcgag gatcgtggca tcaccgaacc gcgccgtgcg cgggtcgtcg
     4801 gtgagccaga gtttcagcag gccgcccagg cggcccaggt cgccattgat gcgggccagc
     4861 tcgcggacgt gctcatagtc cacgacgccc gtgattttgt agccctggcc gacggccagc
     4921 aggtaggcct acaggctcat gccggccgcc gccgcctttt cctcaatcgc tcttcgttcg
     4981 tctggaaggc agtacacctt gataggtggg ctgcccttcc tggttggctt ggtttcatca
     5041 gccatccgct tgccctcatc tgttacgccg gcggtagccg gccagcctcg cagagcagga
     5101 ttcccgttga gcaccgccag gtgcgaataa gggacagtga agaaggaaca cccgctcgcg
     5161 ggtgggccta cttcacctat cctgcccggc tgacgccgtt ggatacacca aggaaagtct
     5221 acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa aggatggata taccgaaaaa
     5281 atcgctataa tgaccccgaa gcagggttat gcagcggaaa agatccgtc
//
