Basic Vector Information
- Vector Name:
- pNS2TrcD
- Antibiotic Resistance:
- Kanamycin
- Length:
- 3114 bp
- Type:
- Expression vector
- Replication origin:
- pBBR1 oriV
- Source/Author:
- Seleem MN, Anderson B, Sriranganathan N.
- Promoter:
- trc
pNS2TrcD vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pNS2TrcD vector Sequence
LOCUS 40924_33522 3114 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pNS2TrcD, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3114)
AUTHORS Seleem MN, Anderson B, Sriranganathan N.
TITLE Improved Expression Vectors for Bartonella
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 3114)
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 3 (bases 1 to 3114)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3114)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(29-MAR-2008) ICTAS, Virginia Tech, 1410 Prices Fork Rd, Blacksburg,
VA 24061, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3114
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 28..57
/label=trc promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 65..81
/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 98..167
/label=rrnG antiterminator
/note="antiterminator from the E. coli rrnG leader region
(Berg et al., 1989)"
CDS 218..241
/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 251..268
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
CDS 600..1406
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDE"
CDS complement(1462..2121)
/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(2122..2891)
/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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