Basic Vector Information
U6p-gRNA1 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 ℃.
U6p-gRNA1 vector Sequence
LOCUS 40924_48987 3493 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector U6p-gRNA1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3493)
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 3493)
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 3493)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3493)
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..3493
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 33..42
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
regulatory 37..46
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
protein_bind 61..85
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
regulatory 90..95
/label=PolIII terminator
/note="PolIII terminator"
/regulatory_class="terminator"
misc_RNA complement(97..172)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(201..441)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
promoter complement(618..636)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(654..670)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(678..694)
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
promoter complement(702..732)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(747..768)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1056..1644)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1818..2675)
/label=AmpR
/note="beta-lactamase"
promoter complement(2676..2780)
/label=AmpR promoter
rep_origin complement(2858..3313)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 3454..3470
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 3477..3493
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
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