LOCUS       Exported                5953 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pR6KT2, complete sequence.
ACCESSION   JX239259
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5953)
  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 5953)
  AUTHORS   Bruckbauer ST.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2012) Microbiology Immunology and Pathology, 
            Colorado State University, IDRC at Foothills Campus, Campus Delivery
            0922, Fort Collins, CO 80523-0922, USA
REFERENCE   3  (bases 1 to 5953)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5953
                     /organism="Cloning vector pR6KT2"
                     /mol_type="other DNA"
                     /db_xref="taxon:1227932"
     primer_bind     269..285
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      436..817
                     /label=R6K oriV
                     /note="R6K oriV"
     rep_origin      441..797
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    841..1102
                     /label=oriT
                     /note="oriT"
     oriT            848..956
                     /note="incP origin of transfer"
     promoter        1306..1334
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             1523..2056
                     /codon_start=1
                     /gene="aacC1"
                     /product="Aminoglycoside N(3)-acetyltransferase"
                     /label=aacC1
                     /note="Confers gentamicin (aminoglycoside) resistance"
                     /protein_id="AFR24687.1"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     gene            1523..2056
                     /gene="aacC1"
                     /label=aacC1
     promoter        2152..2180
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2506..4314
                     /codon_start=1
                     /gene="uidA"
                     /product="beta-D-glucuronidase"
                     /label=uidA
                     /note="Breaks down X-Gluc producing a blue phenotype"
                     /protein_id="AFR24688.1"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQVHGNISPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
                     FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
                     DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
                     KSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            2506..4314
                     /gene="uidA"
                     /label=uidA
     terminator      4371..4416
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(4427..5887)
                     /codon_start=1
                     /gene="sacB"
                     /product="Levansucrase"
                     /label=sacB
                     /note="Counter-selection marker in the presence of sucrose"
                     /protein_id="AFR24689.1"
                     /translation="MTYSKQAVSRRHKRLALSAAALTAAACMSAHAQSDGATQAFAKET
                     NQKPYKETYGISHITRHDMLQIPEQQKNEKYQVSEFDSSTIKNISSAKGLDVWDSWPLQ
                     NADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFD
                     ANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNIN
                     GVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTE
                     DGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKV
                     MKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTG
                     PYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAP
                     SFLLNIKGKKTSVVKDSILEQGQLTVNK"
     gene            complement(4427..5887)
                     /gene="sacB"
                     /label=sacB