Basic Vector Information
- Vector Name:
- pCMV-CBA-Cas9-Rosa26gRNAs
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8986 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Kozhevnikova EN, Leshchenko AE, Pindyurin AV.
- Promoter:
- CBh
pCMV-CBA-Cas9-Rosa26gRNAs vector Vector Map
pCMV-CBA-Cas9-Rosa26gRNAs vector Sequence
LOCUS 40924_11606 8986 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pCMV-CBA-Cas9-Rosa26gRNAs, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8986)
AUTHORS Kozhevnikova EN, Leshchenko AE, Pindyurin AV.
TITLE An inducible DamID system for profiling of a nuclear lamina protein
component Lamin B1 interactions with chromosomes in mouse cells
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8986)
AUTHORS Kozhevnikova EN, Leshchenko AE, Pindyurin AV.
TITLE Direct Submission
JOURNAL Submitted (17-NOV-2017) Department of the Regulation of Genetic
Processes, Institute of Molecular and Cellular Biology, Lavrentyev
ave. 8/2, Novosibirsk, Novosibirsk Region 630090, Russia
REFERENCE 3 (bases 1 to 8986)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8986)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(17-NOV-2017) Department of the Regulation of Genetic Processes,
Institute of Molecular and Cellular Biology, Lavrentyev ave. 8/2,
Novosibirsk, Novosibirsk Region 630090, Russia"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8986
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 70..310
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_feature 319..338
/label=Rosa26 gRNA2 sequence
/note="Rosa26 gRNA2 sequence"
misc_RNA 339..414
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
regulatory 415..420
/note="RNA polymerase III transcription terminator; polIII
terminator"
/regulatory_class="terminator"
regulatory 505..753
/label=human U6 promoter
/note="human U6 promoter"
/regulatory_class="promoter"
promoter 505..745
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
ncRNA 754..855
/product="guide RNA #1; specific to mouse Rosa26 locus"
/label=guide_RNA
/ncRNA_class="guide_RNA"
misc_feature 754..773
/label=Rosa26 gRNA1 sequence
/note="Rosa26 gRNA1 sequence"
misc_feature 774..849
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system; gRNA scaffold"
misc_RNA 774..849
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
regulatory 850..855
/note="RNA polymerase III transcription terminator; polIII
terminator"
/regulatory_class="terminator"
enhancer 939..1224
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 1226..1503
/label=chicken beta-actin promoter
intron 1504..1731
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 1743..1752
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
misc_feature 1752..1817
/label=Region: 3xFLAG
/note="Region: 3xFLAG"
CDS 1752..1817
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
misc_feature 1824..1844
/label=Region: nuclear localization signal (NLS)
/note="Region: nuclear localization signal (NLS)"
CDS 1824..1844
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1869..5969
/label=Cas9
/note="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
misc_feature 5970..6017
/label=Region: nuclear localization signal (NLS)
/note="Region: nuclear localization signal (NLS)"
CDS 5970..6017
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
polyA_signal 6051..6258
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 6267..6407
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
rep_origin 6482..6937
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 7219..7323
/label=AmpR promoter
CDS 7324..8181
/label=AmpR
/note="beta-lactamase"
rep_origin 8355..8943
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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