pNIC-Bio2 vector (V004323)

MSDS

Basic Vector Information

Vector Name:: pNIC-Bio2

Antibiotic Resistance:: Kanamycin

Length:: 7335 bp

Type:: Expression vector

Replication origin:: ori

Source/Author:: 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.

Promoter:: sacB

pNIC-Bio2 vector Vector Map

View Map in NovoBuilder

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 ℃.

pNIC-Bio2 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       40924_33207        7335 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pNIC-Bio2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7335)
  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 29 (5), 515-525 (2012)
  PUBMED    22027370
REFERENCE   2  (bases 1 to 7335)
  AUTHORS   Graslund S, Hammarstrom M, Gileadi O.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-MAY-2011) Structural Genomics Consortium, SGC, 
            University of Oxford, ORCRB, Roosevelt Drive, Oxford OX3 7DQ, UK
REFERENCE   3  (bases 1 to 7335)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7335)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "N 
            Biotechnol"; date: "2012"; volume: "29"; issue: "5"; pages: 
            "515-525"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-MAY-2011) Structural Genomics Consortium, SGC, University of 
            Oxford, ORCRB, Roosevelt Drive, Oxford OX3 7DQ, UK"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7335
                     /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..118
                     /label=9xHis
                     /note="9xHis 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(2084..2097)
                     /note="3'-LIC sequence; add complement to downstream PCR
                     primer"
     CDS             2099..2143
                     /label=AviTag(TM)
                     /note="peptide tag that allows for enzymatic biotinylation"
     CDS             2196..2213
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      2280..2327
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2364..2819
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(2915..3727)
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      3849..4437
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(4623..4765)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(4870..5058)
                     /label=rop
                     /note="Rop protein, which maintains plasmids at low copy
                     number"
     protein_bind    complement(5833..5854)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(5870..6949)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(6950..7027)
                     /label=lacI promoter

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