PL-SIN-PGK-EGFP vector (V007079)

Basic Vector Information

      • Vector Name:
      • PL-SIN-PGK-EGFP
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 6846 bp
      • Type:
      • Mammalian Expression, Lentiviral
      • Copy Number:
      • High Copy
      • Promoter:
      • PGK

PL-SIN-PGK-EGFP vector Vector Map

PL-SIN-PGK-EGFP6846 bp3006009001200150018002100240027003000330036003900420045004800510054005700600063006600pGEX 3'pRS-markerIn lacZ gene5' LTR (truncated)cPPT/CTSPGK promoterKozak sequence3' LTR (Delta-U3)In lacZ genelac promoterCAP binding siteL4440oriAmp-RAmpR promoterSV40 promoterpBRforEco

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 ℃.

PL-SIN-PGK-EGFP vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                6846 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Control Lentiviral vector with ubiquitous PGK promoter, expresses 
            EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    PL-SIN-PGK-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6846)
  AUTHORS   Hotta A, Cheung AY, Farra N, Vijayaragavan K, Seguin CA, Draper JS, 
            Pasceri P, Maksakova IA, Mager DL, Rossant J, Bhatia M, Ellis J
  TITLE     Isolation of human iPS cells using EOS lentiviral vectors to select 
            for pluripotency.
  JOURNAL   Nat Methods. 2009 May . 6(5):370-6.
  PUBMED    19404254
REFERENCE   2  (bases 1 to 6846)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2009 May . 6(5):370-6."
FEATURES             Location/Qualifiers
     source          1..6846
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             785..1042
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1693..1810
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2768..3267
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2803..2826)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3200..3219
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     regulatory      3281..3290
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3287..4006
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3287..4003
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3332..3353)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3593..3612)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3940..3961
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4133..4366
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(4403..4425)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4414..4430
                     /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(4438..4468)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4483..4504
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4621..4638)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4792..5380)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4872..4891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6173..6192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6411..6515)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(6552..6748)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6566..6701
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6620..6639)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6819..6837
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"

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