LOCUS       Exported                7827 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDN19, complete sequence.
ACCESSION   AF327711
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7827)
  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 7827)
  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 7827)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7827
                     /organism="Cloning vector pDN19"
                     /mol_type="other DNA"
                     /note="sequence determined by authors; created for use in 
                     Pseudomonas aeruginosa (Nunn et al., J. Bacteriol. 172, 
                     2911-2919 (1990)"
                     /db_xref="taxon:163910"
     rep_origin      4..712
                     /note="oriV; IncP origin of replication"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     CDS             complement(2576..2842)
                     /codon_start=1
                     /gene="lacZa"
                     /product="LacZ"
                     /label=lacZa
                     /note="beta-galactosidase alpha peptide"
                     /protein_id="AAK73367.1"
                     /translation="MTMITPSSNTTHYRESLHACRSTLEDPRVPSSNSLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWKL"
     gene            complement(2576..2842)
                     /gene="lacZa"
                     /label=lacZa
     primer_bind     2725..2741
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2742..2798
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(2801..2819)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(2804..2819)
                     /regulatory_class="promoter"
                     /note="PT7; T7 promoter"
     primer_bind     complement(2838..2854)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(2861..2916)
                     /regulatory_class="promoter"
                     /note="Plac; lac operon promoter/operator"
     protein_bind    2862..2878
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     promoter        complement(2886..2916)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2931..2952
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             3280..4479
                     /codon_start=1
                     /gene="tetA"
                     /product="TetA"
                     /function="tetracycline resistance"
                     /label=tetA
                     /protein_id="AAK73368.1"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     gene            3280..4479
                     /gene="tetA"
                     /label=tetA
     CDS             complement(5752..6900)
                     /codon_start=1
                     /gene="trfA"
                     /product="TrfA"
                     /label=trfA
                     /note="essential replication protein"
                     /protein_id="AAK73369.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(5752..6900)
                     /gene="trfA"
                     /label=trfA
     oriT            7172..7543
                     /label=IncP origin of transfer
                     /note="IncP origin of transfer"
     CDS             complement(7172..7543)
                     /codon_start=1
                     /gene="traJ"
                     /product="TraJ"
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /protein_id="AAK73370.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     gene            complement(7172..7543)
                     /gene="traJ"
                     /label=traJ
     oriT            7576..7685
                     /note="incP origin of transfer"