U6p-gRNA1 vector (V001584)

Basic Vector Information

      • Vector Name:
      • U6p-gRNA1
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 3493 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK.

U6p-gRNA1 vector Vector Map

U6p-gRNA13493 bp6001200180024003000vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)attB2PolIII terminatorgRNA1SP6 promoterM13 revlac operatorlac promoterCAP binding siteoriampicillin resistance proteinAmpR promoterf1 oriM13 fwd

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                3493 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector U6p-gRNA1, complete sequence.
ACCESSION   KJ796490
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3493
                     /organism="Cloning vector U6p-gRNA1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1520553"
     regulatory      33..42
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      37..46
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     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
                     /regulatory_class="terminator"
                     /note="PolIII terminator"
     misc_feature    96..191
                     /label=gRNA1
                     /note="gRNA1"
     misc_RNA        complement(97..172)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      193..510
                     /regulatory_class="promoter"
                     /gene="U6"
     gene            193..510
                     /gene="U6"
                     /label=U6
     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    678..694
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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    747..768
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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(1815..2675)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="AIJ03089.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2676..2780)
                     /gene="bla"
                     /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..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"

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