Basic Vector Information
- Vector Name:
- pNICKclos1.0
- Antibiotic Resistance:
- Ampicillin
- Length:
- 11082 bp
- Type:
- Gene knockout
- Replication origin:
- ori
- Host:
- Clostridium
- Selection Marker:
- Ery
- Promoter:
- J23119(SpeI)
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pNICKclos1.0 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 ℃.
pNICKclos1.0 vector Sequence
LOCUS 62056_18020 11082 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11082)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..11082
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 14..48
/label=J23119(SpeI) promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119) modified to end with an SpeI site"
misc_RNA 69..144
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 2304..6407
/label=Cas9(D10A)
/note="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
promoter 6912..7016
/label=AmpR promoter
CDS 7017..7874
/label=AmpR
/note="beta-lactamase"
rep_origin 8048..8636
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 8924..8945
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 8960..8990
/label=lac promoter
/note="promoter for the E. coli lac operon"
CDS 9406..10137
/gene="ermC'"
/label=ermC'
/note="rRNA adenine N-6-methyltransferase from Bacillus
subtilis. Accession#: P13956"
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