Basic Vector Information
CMVp-HHRibo-gRNA1-HDVRibo-pA 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 ℃.
CMVp-HHRibo-gRNA1-HDVRibo-pA vector Sequence
LOCUS 40924_505 3928 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector CMVp-HHRibo-gRNA1-HDVRibo-pA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3928)
AUTHORS Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK.
TITLE Multiplexed and programmable regulation of gene networks with an
integrated RNA and CRISPR/Cas toolkit in human cells
JOURNAL Mol. Cell 54 (4), 698-710 (2014)
PUBMED 24837679
REFERENCE 2 (bases 1 to 3928)
AUTHORS Perli SD.
TITLE Direct Submission
JOURNAL Submitted (04-MAY-2014) Electrical Engineering and Computer Science,
Massachusetts Institute of Technology, 77, Massachusetts Avenue,
Cambridge, MA 02139, USA
REFERENCE 3 (bases 1 to 3928)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3928)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Cell";
date: "2014"; volume: "54"; issue: "4"; pages: "698-710"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(04-MAY-2014) Electrical Engineering and Computer Science,
Massachusetts Institute of Technology, 77, Massachusetts Avenue,
Cambridge, MA 02139, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3928
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(620..724)
/label=AmpR promoter
rep_origin 751..1206
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
enhancer 1395..1698
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1699..1902
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
ncRNA 1936..1972
/label=hammerhead ribozyme
misc_RNA 1993..2068
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
ncRNA 2073..2140
/label=HDV ribozyme
polyA_signal 2389..2436
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin complement(2928..3516)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(join(3690..3928,1..619))
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
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