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P1-EYFP-pA vector (V009798#)

Basic Vector Information

Basic Vector Information
Vector Name P1-EYFP-pA Antibiotic Resistance Ampicillin
Length 4215 bp Type Cloning vector
Source Nissim L, Perli SD, Fridkin A, Perez-Pinera P, Lu TK.

P1-EYFP-pA vector Vector Map

ampicillin resistance protein AmpR promoter f1 ori synthetic polyadenylation signal pause site 8x gRNA1 binding sites Minimal Ade MLP PhiYFP SV40

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 ℃.

P1-EYFP-pA vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                4215 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector P1-EYFP-pA, complete sequence.
ACCESSION   KJ796488
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4215)
  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 4215)
  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 4215)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4215
                     /organism="Cloning vector P1-EYFP-pA"
                     /mol_type="other DNA"
                     /db_xref="taxon:1520550"
     CDS             complement(468..1328)
                     /codon_start=1
                     /product="ampicillin resistance protein"
                     /label=ampicillin resistance protein
                     /protein_id="AIJ03087.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1329..1433)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1460..1915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    2046..2094
                     /note="synthetic polyadenylation signal"
     misc_feature    2108..2199
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     misc_feature    2224..2574
                     /label=8x gRNA1 binding sites
                     /note="8x gRNA1 binding sites"
     regulatory      2581..2614
                     /regulatory_class="promoter"
                     /note="Minimal Ade MLP"
     CDS             2649..3353
                     /codon_start=1
                     /product="Phialidium yellow fluorescent proteim"
                     /label=PhiYFP
                     /note="mammalian codon-optimized"
                     /translation="MSSGALLFHGKIPYVVEMEGDVDGHTFSIRGKGYGDASVGKVDAQ
                     FICTTGDVPVPWSTLVTTLTYGAQCFAKYGPELKDFYKSCMPDGYVQERTITFEGDGNF
                     KTRAEVTFENGSVYNRVKLNGQGFKKDGHVLGKNLEFNFTPHCLYIWGDQANHGLKSAF
                     KICHEIAGSKGDFIVADHTQMNTPIGGGPVHVPEYHHMSYHVKLSKDVTDHRDNMSLTE
                     TVRAVDCRKTYL"
     gene            2649..3350
                     /gene="EYFP"
                     /label=EYFP
     regulatory      3370..3591
                     /regulatory_class="polyA_signal_sequence"
                     /gene="SV40"
     gene            3370..3591
                     /gene="SV40"
                     /label=SV40
     polyA_signal    3379..3500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3924..297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"

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