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pwtCas9-bacteria vector (V012764)

Basic Vector Information

aTc-inducible expression of wild-type Cas9 (S .pyogenes) for bacterial gene knockdown

Vector Name:
pwtCas9-bacteria
Antibiotic Resistance:
Ampicillin
Length:
6932 bp
Type:
CRISPR Plasmids
Replication origin:
ori
Source/Author:
Stanley Qi
Copy Number:
High copy number
Promoter:
tetR/tetAs
Expression Method:
aTc-inducible

pwtCas9-bacteria vector Vector Map

pwtCas9-bacteria6932 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900TetRtetR/tetA promotersRBSCas9rrnB T1 terminatorT7Te terminatororilambda t0 terminatorAmpRAmpR 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 ℃.

pwtCas9-bacteria vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pwtCas9-bacteria        6932 bp DNA     circular SYN 18-SEP-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pwtCas9-bacteria.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6932)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6932)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(10..633)
                     /label=TetR
                     /note="tetracycline repressor TetR"
     promoter        649..704
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     RBS             722..733
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             740..4843
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     terminator      4870..4941
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4957..4984
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(5151..5739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(5827..5921)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(5954..6811)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6812..6916)
                     /label=AmpR promoter

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