CMVp-HHRibo-gRNA1-HDVRibo-pA vector (V010035#)

Basic Vector Information

Basic Vector Information
Vector Name CMVp-HHRibo-gRNA1-HDVRibo-pA Antibiotic Resistance Ampicillin
Length 3928 bp Type Cloning vector
Source Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK.

CMVp-HHRibo-gRNA1-HDVRibo-pA vector Vector Map

CMVp-HHRibo-gRNA1-HDVRibo-pA3928 bp60012001800240030003600AmpR promoterf1 oripmCMV5_prime HH self-cleaving ribozymegRNA13_prime HDV self-cleaving ribozymeoriampicillin resistance protein

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

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                3928 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector CMVp-HHRibo-gRNA1-HDVRibo-pA, complete sequence.
ACCESSION   KJ796498
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3928
                     /organism="Cloning vector CMVp-HHRibo-gRNA1-HDVRibo-pA"
                     /mol_type="other DNA"
                     /db_xref="taxon:1520530"
     promoter        complement(620..724)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      751..1206
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     regulatory      1335..1923
                     /regulatory_class="promoter"
                     /note="pmCMV"
     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"
     misc_feature    1930..1972
                     /label=5_prime HH self-cleaving ribozyme
                     /note="5_prime HH self-cleaving ribozyme"
     misc_feature    1973..2072
                     /label=gRNA1
                     /note="gRNA1"
     misc_RNA        1993..2068
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    2073..2140
                     /label=3_prime HDV self-cleaving ribozyme
                     /note="3_prime HDV self-cleaving ribozyme"
     regulatory      2158..2607
                     /regulatory_class="polyA_signal_sequence"
                     /gene="HSV"
     gene            2158..2607
                     /gene="HSV"
                     /label=HSV
     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(3687..619)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="AIJ03097.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"

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