LOCUS       Exported                7965 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pCM130, complete sequence.
ACCESSION   AF327719
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7965)
  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 7965)
  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 7965)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7965
                     /organism="Cloning vector pCM130"
                     /mol_type="other DNA"
                     /note="low-background broad-host-range promoter-probe 
                     vector using xylE as a reporter gene; created by inserting 
                     a 1.0 kb fragment containing xylE (from pCM75) into pCM62 
                     cut with AflIII and HindIII, followed by the insertion of a
                     0.5 kb XbaI-XmnI trrnB fragment from pMTL20T1T2 into the 
                     blunted EcoRI site"
                     /db_xref="taxon:163918"
     rep_origin      4..712
                     /note="oriV; IncP origin of replication"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      1180..1735
                     /label=colE1
                     /note="colE1"
     CDS             complement(1770..2693)
                     /codon_start=1
                     /gene="xylE"
                     /product="XylE"
                     /label=xylE
                     /note="catechol 2,3-dioxygenase"
                     /protein_id="AAK73418.1"
                     /translation="MNKGVMRPGHVQLRVLDMSKALEHYVELLGLIEMDRDDQGRVYLK
                     AWTEVDKFSLVLREADEPGMDFMGFKVVDEDALRQLERDLMAYGCAVEQLPAGELNSCG
                     RRVRFQAPSGHHFELYADKEYTGKWGLNDVNPEAWPRDLKGMAAVRFDHALMYGDELPA
                     TYDLFTKVLGFYLAEQVLDENGTRVAQFLSLSTKAHDVAFIHHPEKGRLHHVSFHLETW
                     EDLLRAADLISMTDTSIDIGPTRHGLTHGKTIYFFDPSGNRNEVFCGGDYNYPDHKPVT
                     WTTDQLGKAIFYHDRILNERFMTVLT"
     gene            complement(1770..2693)
                     /gene="xylE"
                     /label=xylE
     regulatory      2783..3246
                     /regulatory_class="terminator"
                     /note="trrnB from Escherichia coli"
     terminator      2865..2892
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      2984..3070
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3252..3268)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3689..4354)
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /function="tetracycline resistance regulation"
                     /label=tetR
                     /protein_id="AAK73419.1"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGAKKPLLS"
     gene            complement(3689..4354)
                     /gene="tetR"
                     /label=tetR
     CDS             4460..5659
                     /codon_start=1
                     /gene="tetA"
                     /product="TetA"
                     /function="tetracycline resistance"
                     /label=tetA
                     /protein_id="AAK73420.1"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     gene            4460..5659
                     /gene="tetA"
                     /label=tetA
     CDS             complement(5890..7038)
                     /codon_start=1
                     /gene="trfA"
                     /product="TrfA"
                     /function="trans-acting replication"
                     /label=trfA
                     /protein_id="AAK73421.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(5890..7038)
                     /gene="trfA"
                     /label=trfA
     oriT            7220..7958
                     /label=IncP origin of transfer
                     /note="IncP origin of transfer"
     CDS             complement(7310..7681)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             complement(7565..7681)
                     /codon_start=1
                     /gene="traJ"
                     /product="TraJ"
                     /label=traJ
                     /note="truncated oriT-recognizing protein"
                     /protein_id="AAK73422.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL"
     gene            complement(7565..7681)
                     /gene="traJ"
                     /label=traJ
     oriT            7714..7823
                     /note="incP origin of transfer"