pNS2Trc vector (V004276)

Basic Vector Information

Vector Name:
pNS2Trc
Antibiotic Resistance:
Kanamycin
Length:
3109 bp
Type:
Expression vector
Replication origin:
pBBR1 oriV
Source/Author:
Gillaspie D, Perkins I, Larsen K, McCord A, Pangonis S, Sweger D, Seleem MN, Sriranganathan N, Anderson BE.
Promoter:
trc

pNS2Trc vector Map

pNS2Trc3109 bp6001200180024003000trc promoterlac operatorrrnG antiterminatorminicistron6xHisKanRpBBR1 ReppBBR1 oriV

pNS2Trc vector Sequence

LOCUS       40924_33512        3109 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNS2Trc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3109)
  AUTHORS   Gillaspie D, Perkins I, Larsen K, McCord A, Pangonis S, Sweger D, 
            Seleem MN, Sriranganathan N, Anderson BE.
  TITLE     Plasmid-based system for high-level gene expression and antisense 
            gene knockdown in Bartonella henselae
  JOURNAL   Appl. Environ. Microbiol. 75 (16), 5434-5436 (2009)
  PUBMED    19542333
REFERENCE   2  (bases 1 to 3109)
  AUTHORS   Seleem MN, Anderson B, Sriranganathan N.
  TITLE     Improved Expression Vectors for Bartonella
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 3109)
  AUTHORS   Seleem MN, Anderson B, Sriranganathan N.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-MAR-2008) ICTAS, Virginia Tech, 1410 Prices Fork Rd, 
            Blacksburg, VA 24061, USA
REFERENCE   4  (bases 1 to 3109)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 3109)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2009"; volume: "75"; issue: "16"; 
            pages: "5434-5436"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (29-MAR-2008) ICTAS, Virginia Tech, 1410 Prices Fork Rd, Blacksburg,
            VA 24061, USA"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        23..52
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    60..76
                     /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)."
     misc_feature    93..162
                     /label=rrnG antiterminator
                     /note="antiterminator from the E. coli rrnG leader region
                     (Berg et al., 1989)"
     CDS             213..236
                     /codon_start=1
                     /label=minicistron
                     /note="synthetic cistron containing a ribosome binding site
                     (Shine-Dalgarno sequence), for enhancing the bacterial 
                     expression of a downstream cistron (Schoner, 1997)"
                     /translation="MYRLNKEE"
     CDS             246..263
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             595..1401
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDE"
     CDS             complement(1457..2116)
                     /codon_start=1
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      complement(2117..2886)
                     /direction=LEFT
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"

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