Basic Vector Information
- Vector Name:
- CMV-dCas13X.1-RESCUE-S-SV40pA_U6-BbsI-DR_CMV-mCherry-P2A-Puro-BGHpA
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9693 bp
- Type:
- Genome editing
- Replication origin:
- ori
- Host:
- Mammalian cells
- Promoter:
- CBh
- Growth Temperature:
- 37℃
CMV-dCas13X.1-RESCUE-S-SV40pA_U6-BbsI-DR_CMV-mCherry-P2A-Puro-BGHpA 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 ℃.
CMV-dCas13X.1-RESCUE-S-SV40pA_U6-BbsI-DR_CMV-mCherry-P2A-Puro-BGHpA vector Sequence
LOCUS 62056_546 9693 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 9693)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9693
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 396..681
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 683..960
/label=chicken beta-actin promoter
CDS 1209..1229
/codon_start=1
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
CDS 3552..3572
/codon_start=1
/label=SV40 NLS
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/translation="PKKKRKV"
polyA_signal 4800..4934
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 4941..5181
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
enhancer 5331..5634
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5635..5838
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 5873..6580
/codon_start=1
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
DDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
CDS 6647..7240
/codon_start=1
/label=PuroR
/note="puromycin N-acetyltransferase"
/translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
polyA_signal 7250..7474
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter 7896..8000
/label=AmpR promoter
CDS 8001..8858
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 9032..9620
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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