P2-ECFP-pA vector (V009774)

Basic Vector Information

      • Vector Name:
      • P2-ECFP-pA
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 4180 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK.

P2-ECFP-pA vector Vector Map

P2-ECFP-pA4180 bp60012001800240030003600ampicillin resistance proteinAmpR promoterf1 oripause site8x gRNA2 binding sitesMinimal Ade MLPmCerulean

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 ℃.

P2-ECFP-pA vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                4180 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector P2-ECFP-pA, complete sequence.
ACCESSION   KJ796489
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4180)
  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 4180)
  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 4180)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4180
                     /organism="Cloning vector P2-ECFP-pA"
                     /mol_type="other DNA"
                     /db_xref="taxon:1520551"
     CDS             complement(448..1308)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="AIJ03088.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1309..1413)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1440..1895
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     misc_feature    2087..2178
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     misc_feature    2186..2511
                     /label=8x gRNA2 binding sites
                     /note="8x gRNA2 binding sites"
     regulatory      2518..2549
                     /regulatory_class="promoter"
                     /note="Minimal Ade MLP"
     CDS             2575..3294
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            2575..3291
                     /gene="ECFP"
                     /label=ECFP
     regulatory      3311..3532
                     /regulatory_class="polyA_signal_sequence"
                     /gene="SV40"
     gene            3311..3532
                     /gene="SV40"
                     /label=SV40
     polyA_signal    3320..3441
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3869..277)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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