LOCUS       Exported                5329 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCM51, complete sequence.
ACCESSION   AF327713
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5329
                     /organism="Cloning vector pCM51"
                     /mol_type="other DNA"
                     /note="minimal transferable and selectable IncP replicon 
                     created from pDN19X, a spontaneous mutant of pDN19; served 
                     as the IncP backbone for the construction of a series of 
                     improved broad-host-range vectors"
                     /db_xref="taxon:163912"
     rep_origin      4..712
                     /note="oriV; IncP origin of replication"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     CDS             complement(1068..1718)
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /function="tetracycline resistance regulation"
                     /label=tetR
                     /protein_id="AAK73392.1"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     gene            complement(1068..1718)
                     /gene="tetR"
                     /label=tetR
     CDS             1824..3023
                     /codon_start=1
                     /gene="tetA"
                     /product="TetA"
                     /function="tetracycline resistance"
                     /label=tetA
                     /protein_id="AAK73393.1"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     gene            1824..3023
                     /gene="tetA"
                     /label=tetA
     CDS             complement(3254..4402)
                     /codon_start=1
                     /gene="trfA"
                     /product="TrfA"
                     /function="trans-acting replication"
                     /label=trfA
                     /protein_id="AAK73394.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(3254..4402)
                     /gene="trfA"
                     /label=trfA
     oriT            4584..5322
                     /label=IncP origin of transfer
                     /note="IncP origin of transfer"
     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            5078..5187
                     /note="incP origin of transfer"