LOCUS       Exported                7299 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Pasteurellaceae shuttle vector pMK-Express, complete sequence.
ACCESSION   GQ334690
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7299)
  AUTHORS   Bosse JT, Durham AL, Rycroft AN, Kroll JS, Langford PR.
  TITLE     New plasmid tools for genetic analysis of Actinobacillus 
            pleuropneumoniae and other pasteurellaceae
  JOURNAL   Appl. Environ. Microbiol. 75 (19), 6124-6131 (2009)
  PUBMED    19666733
REFERENCE   2  (bases 1 to 7299)
  AUTHORS   Bosse JT, Durham AL, Rycroft AN, Kroll JS, Langford PR.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-JUN-2009) Paediatrics, Imperial College London, 
            Norfolk Place, London W2 1PG, UK
REFERENCE   3  (bases 1 to 7299)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7299
                     /organism="Pasteurellaceae shuttle vector pMK-Express"
                     /mol_type="other DNA"
                     /db_xref="taxon:656015"
     misc_feature    53..261
                     /note="contains Actinobacillus pleuropneumoniae sodC 
                     promoter"
     misc_feature    262..304
                     /note="synthetic polylinker cloning domain; contains 
                     restriction sites EcoRI, PstI, KpnI, ApaI, BamHI, and NheI"
     CDS             332..1048
                     /codon_start=1
                     /gene="GFPmut3"
                     /product="GFPmut3"
                     /label=GFPmut3
                     /note="green fluorescent protein mut3 variant"
                     /protein_id="ACT56510.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            332..1048
                     /gene="GFPmut3"
                     /label=GFPmut3
     misc_feature    1088..1173
                     /note="synthetic polylinker cloning domain; contains 
                     restriction sites XbaI, SacI, BstXI, NotI, and a second 
                     XbaI site"
     primer_bind     1094..1110
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1117..1135
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1199..1215)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1223..1239
                     /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(1247..1277)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1292..1313
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(1921..3357)
                     /codon_start=1
                     /gene="mobA"
                     /product="MobA"
                     /label=mobA
                     /note="mobilization protein A; MobA/MobL family; pfam03389"
                     /protein_id="ACT56511.1"
                     /translation="MAIYHLNVRYCSKSKGQSAQAKNDYINRNDKYSKRLDDLQFSGYG
                     NMPKFAEDNPQEFWRLSDIYERANARVCTEIEFALPRELTLEQQQKLVSSFIENTVDSG
                     SNKLPYSFAIHTDKNNHNPHCHLIFSERQLDGIDRTAEQFFKRANTKSPEKGGAMKTAD
                     FRDREFIQSVRKTWREQANQALEQYGYAARIDERSYKEQGIEQAPRARIDRVTWQELNR
                     LEQEERQIVQELALKGQEINKEKSYLQKIEEKQAQGMGKYESKFAAAFSKLSESALKHD
                     LSNEKEKDSKIHTQEEKVPQNRIQGLSQADFDQFLIDEWLPQIEKYVKAQEKRDGMEVE
                     ITQYDKDLQRIQGDYNKLTDKNQGFLGLWETKEQKAKKKELEDEYKHTAEQRNAKSQEL
                     AEYSQKIKAYEQKTLEPINEKIAKYQADNPEIKMRSLGFVKKIKAQGAYKAAQERMERE
                     KQHQQEKQQRHLERESGLSL"
     gene            complement(1921..3357)
                     /gene="mobA"
                     /label=mobA
     CDS             complement(5994..6788)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="AphA-3"
                     /label=aphA-3
                     /note="aminoglycoside phosphotransferase; kanamycin 
                     resistance cassette"
                     /protein_id="ACT56512.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCIRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(5994..6788)
                     /gene="aphA-3"
                     /label=aphA-3