VEE(wt).Pac-2A-GFP vector (V001513)

Basic Vector Information

      • Vector Name:
      • VEE(wt).Pac-2A-GFP
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 14800 bp
      • Type:
      • Cloning vector
      • Source/Author:
      • Kim YG, Baltabekova AZ, Shustov AV.

VEE(wt).Pac-2A-GFP vector Vector Map

VEE(wt).Pac-2A-GFP14800 bp700140021002800350042004900560063007000770084009100980010500112001190012600133001400014700RBSPuroREGFPAmpR promoterAmpRori

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 ℃.

VEE(wt).Pac-2A-GFP vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported               14800 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector VEE(wt).Pac-2A-GFP, complete sequence.
ACCESSION   MF136451
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14800)
  AUTHORS   Kim YG, Baltabekova AZ, Shustov AV.
  TITLE     Poxvirus' interferon inhibitor B18R: recombinant expression, 
            refolding and use in mammalian expression system based on viral 
            vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14800)
  AUTHORS   Kim YG, Baltabekova AZ, Shustov AV.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-MAY-2017) National Center for Biotechnology, 
            Korgalzhin hwy 13/5, Astana, Akmola region 010000, Kazakhstan
REFERENCE   3  (bases 1 to 14800)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..14800
                     /organism="Cloning vector VEE(wt).Pac-2A-GFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:2022541"
     RBS             1536..1544
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             7576..8172
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     CDS             8224..8943
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     promoter        13038..13142
                     /gene="bla"
                     /label=AmpR promoter
     CDS             13143..14003
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      14174..14762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        14784..2
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"

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