pNIC28-Bsa4 vector (V010577)

Price Information

Cat No. Plasmid Name Availability Add to cart
V010577 pNIC28-Bsa4 In stock (lyophilized plasmid)

Buy one, get one free!

Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.

Basic Vector Information

Vector Name:
pNIC28-Bsa4
Antibiotic Resistance:
Kanamycin
Length:
7284 bp
Type:
Structural Genomics Vectors
Replication origin:
ori
Source/Author:
Savitsky P, Bray J, Cooper CD, Marsden BD, Mahajan P, Burgess-Brown
Copy Number:
High copy number
Promoter:
sacB

pNIC28-Bsa4 vector Vector Map

pNIC28-Bsa47284 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200T7 promoterlac operatorRBS6xHisTEV sitesacB promoterSacB3'-LIC sequence; add complement to downstream PCR primer6xHisT7 terminatorf1 oriKanRoribomropCAP binding sitelacIlacI promoter

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 ℃.

pNIC28-Bsa4 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_33252        7284 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNIC28-Bsa4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7284)
  AUTHORS   Stols L, Gu M, Dieckman L, Raffen R, Collart FR, Donnelly MI.
  TITLE     A new vector for high-throughput, ligation-independent cloning 
            encoding a tobacco etch virus protease cleavage site
  JOURNAL   Protein Expr. Purif. 25 (1), 8-15 (2002)
  PUBMED    12071693
REFERENCE   2  (bases 1 to 7284)
  AUTHORS   Keates T, Cooper CD, Savitsky P, Allerston CK, Phillips C, 
            Hammarstrom M, Daga N, Berridge G, Mahajan P, Burgess-Brown NA, 
            Muller S, Graslund S, Gileadi O.
  TITLE     Expressing the human proteome for affinity proteomics: optimising 
            expression of soluble protein domains and in vivo biotinylation
  JOURNAL   N Biotechnol (2011) In press
  PUBMED    22027370
REFERENCE   3  (bases 1 to 7284)
  AUTHORS   Gileadi O, Burgess-Brown N, Loppnau P.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-DEC-2006) Structural Genomics Consortium, University 
            of Oxford, Botnar Research Centre, Oxford OX3 7LD, UK
REFERENCE   4  (bases 1 to 7284)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7284)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Protein 
            Expr. Purif."; date: "2002"; volume: "25"; issue: "1"; pages: "8-15"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "N 
            Biotechnol (2011) In press"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (25-DEC-2006) Structural Genomics Consortium, University of Oxford, 
            Botnar Research Centre, Oxford OX3 7LD, UK"
COMMENT     SGRef: number: 4; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7284
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /label=6xHis
                     /note="6xHis affinity tag"
     CDS             134..154
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
     promoter        170..615
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             616..2034
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     misc_feature    complement(2083..2097)
                     /note="3'-LIC sequence; add complement to downstream PCR
                     primer"
     CDS             2145..2162
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      2229..2276
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2313..2768
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2864..3676)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      3798..4386
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4572..4714)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4819..5007)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(5782..5803)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5819..6898)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(6899..6976)
                     /label=lacI promoter