LOCUS       Exported                7427 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Plasmid vector pEXGm5B, complete sequence.
ACCESSION   JX504716
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7427)
  AUTHORS   Kvitko BH, Bruckbauer S, Prucha J, McMillan I, Breland EJ, Lehman S,
            Mladinich K, Choi KH, Karkhoff-Schweizer R, Schweizer HP.
  TITLE     A simple method for construction of pir+ Enterobacterial hosts for 
            maintenance of R6K replicon plasmids
  JOURNAL   BMC Res Notes 5, 157 (2012)
  PUBMED    22433797
REFERENCE   2  (bases 1 to 7427)
  AUTHORS   Kvitko BH, Goodyear A, Propst KL, Dow SW, Schweizer HP.
  TITLE     Burkholderia pseudomallei Known Siderophores and Hemin Uptake Are 
            Dispensable for Lethal Murine Melioidosis
  JOURNAL   PLoS Negl Trop Dis 6 (6), E1715 (2012)
  PUBMED    22745846
REFERENCE   3  (bases 1 to 7427)
  AUTHORS   Kvitko BH, Schweizer HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-AUG-2012) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   4  (bases 1 to 7427)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7427
                     /organism="Plasmid vector pEXGm5B"
                     /mol_type="other DNA"
                     /db_xref="taxon:1245208"
     misc_feature    12..836
                     /label=RP4 oriT
                     /note="RP4 oriT"
     oriT            358..467
                     /note="incP origin of transfer"
     terminator      892..919
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      1011..1097
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1253..1558)
                     /codon_start=1
                     /product="LacZ alpha polypeptide"
                     /label=LacZ alpha polypeptide
                     /note="LacZ alpha-complementation polypeptide for blue 
                     white screening on X-gal; lacZ alpha"
                     /protein_id="AFV53144.1"
                     /translation="MTMITPSSRDNRVIPNSRGGRPGRDNRVMSLALAVVLQRRDWENP
                     GVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRAWLFWRMREDFQPDTD"
     primer_bind     1447..1463
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1554..1570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1578..1594
                     /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(1602..1632)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1647..1668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      1874..2547
                     /label=pUC origin
                     /note="pUC origin"
     rep_origin      complement(1956..2544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        2780..2808
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2997..3530
                     /codon_start=1
                     /gene="aacCI"
                     /product="AacCI"
                     /EC_number="2.3.1.81"
                     /label=aacCI
                     /note="aminoglycoside 3-acetyltransferase; gentamicin 
                     resistance"
                     /protein_id="AFV53143.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            2997..3530
                     /gene="aacCI"
                     /label=aacCI
     promoter        3626..3654
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3980..5788
                     /codon_start=1
                     /gene="uidA"
                     /gene_synonym="gusA"
                     /product="UidA"
                     /EC_number="3.2.1.31"
                     /label=uidA
                     /note="beta-glucuronidase; creates blue precipitate on 
                     X-gluc"
                     /protein_id="AFV53142.1"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQVHGNISPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
                     FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
                     DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
                     KSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            3980..5788
                     /gene="uidA"
                     /gene_synonym="gusA"
                     /label=uidA
     terminator      5845..5890
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5901..7361)
                     /codon_start=1
                     /gene="sacB"
                     /product="SacB"
                     /EC_number="2.4.1.10"
                     /label=sacB
                     /note="sucrose counter-selection; levansucrase"
                     /protein_id="AFV53141.1"
                     /translation="MTYSKQAVSRRHKRLALSAAALTAAACMSAHAQSDGATQAFAKET
                     NQKPYKETYGISHITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQ
                     NADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFD
                     ANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNIN
                     GVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTE
                     DGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKV
                     MKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTG
                     PYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAP
                     SFLLNIKGKKTSVVKDSILEQGQLTVNK"
     gene            complement(5901..7361)
                     /gene="sacB"
                     /label=sacB