pET28A-blaSPM-1 vector (V015924)

LOCUS       62056_9965        6397 bp DNA     circular SYN 06-MAY-2019
DEFINITION  Cloning vector pET28A-blaSPM-1, complete sequence.
ACCESSION   MK847895
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6397)
  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  JOURNAL   Unpublished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 6397)
  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 6397)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     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"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6397
                     /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"
     gene            5273..6103
                     /gene="blaSPM"
                     /allele="blaSPM-1"
                     /label=blaSPM
     CDS             5273..6103
                     /codon_start=1
                     /transl_table=11
                     /gene="blaSPM"
                     /allele="blaSPM-1"
                     /product="subclass B1 metallo-beta-lactamase SPM-1"
                     /label=blaSPM
                     /protein_id="QCH41439.1"
                     /translation="MNSPKSRALLGFMGAFCLLLVAGAPLSAKSSDHVDLPYNLTATKI
                     DSDVFVVTDRDFYSSNVLVAKMLDGTVVIVSSPFENLGTQTLMDWVAKTMKPKKVVAIN
                     THFHLDGTGGNEIYKKMGAETWSSDLTKQLRLEENKKDRIKAAEFYKNEDLKRRILSSH
                     PVPADNVFDLKQGKVFSFSNELVEVSFPGPAHSPDNVVVYFPKKKLLFGGCMIKPKELG
                     YLGDANVKAWPDSARRLKKFDAKIVIPGHGEWGGPEMVNKTIKVAEKAVGEMRL"
     CDS             6241..6258
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      6325..6372
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"