pET28A-blaNMC-A vector (V016064)

LOCUS       V016064                 7335 bp    DNA     circular SYN 06-MAY-2019
DEFINITION  Exported.
ACCESSION   V016064
VERSION     V016064
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7335)
  AUTHORS   Hu XM, Yan XH, Yang B, Wang Q, Zheng L.
  TITLE     Full-length Real-time PCR for Rapid Detection of Seventeen
            Carbapenemase Genes in Gram-negative bacteria
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7335)
  AUTHORS   Hu XM, Yan XH, Yang B, Wang Q, Zheng L.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-APR-2019) Department of Laboratory Medicine, Nanfang
            Hospital, Southern Medical University, No. 1383, Guangzhou Dadao
            Bei, Guangzhou, Guangdong 510515, China
REFERENCE   3  (bases 1 to 7335)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (25-APR-2019) Department of Laboratory Medicine, Nanfang Hospital,
            Southern Medical University, No. 1383, Guangzhou Dadao Bei,
            Guangzhou, Guangdong 510515, China"
FEATURES             Location/Qualifiers
     source          1..7335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(563..1375)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      1497..2085
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     misc_feature    complement(2271..2413)
                     /label="bom"
                     /note="basis of mobility region from pBR322"
     CDS             complement(2518..2706)
                     /label="rop"
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(3481..3502)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3518..4597)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(4598..4675)
                     /label="lacI promoter"
     promoter        4984..5002
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5003..5027
                     /label="lac operator"
                     /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)."
     RBS             5042..5064
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5083..5100
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             5110..5127
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             5131..5163
                     /label="T7 tag (gene 10 leader)"
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             complement(5190..6077)
                     /codon_start=1
                     /transl_table=11
                     /product="transcriptional regulator NMC-R"
                     /label="transcriptional regulator NMC-R"
                     /protein_id="QCH41444.1"
                     /translation="MRARLPLNALRAFEASARYLNFTKAGLELHVSQAAVSQQVRTLEQ
                     MLGVALFTRVPRGLQLTDEGMHLLPSITEALQMMSSAMDKFHEGKIKEVLTIAVVGTFA
                     IGWLLPRITAFLNENPWIDIRILTHNNVVNLAAEGIDASIRFGTGGWINTENILLFQAP
                     HTVLCSPETSKKLYIPSDLKKVCLLRSYRKEEWNNWFKAAGIDPWTITGPIFDSTRLMI
                     DAVKLGDYAALVPYHMFQKELNERSVAKPFEIYATLGGYWLTLQKSRVNHNSEALNVFK
                     EWIIEHSREFVLKS"
     CDS             6219..7094
                     /gene="nmcA"
                     /label="Imipenem-hydrolyzing beta-lactamase"
                     /note="Imipenem-hydrolyzing beta-lactamase from
                     Enterobacter cloacae. Accession#: P52663"
     CDS             7179..7196
                     /label="6xHis"
                     /note="6xHis affinity tag"
     terminator      7263..7310
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"