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PVT2339        2ug


pRRLSIN.CPPT.PGK-GFP.WPRE plasmid infomation

Promoters: hPGK, T3, T7
Replicon: pUC ori, F1 ori, SV40 ori
Terminator: SV40 poly (A) signal
Plasmid classification: virus series, lentiviral cloning vector
Prokaryotic resistance: Amp
Clone strain: Stbl3
Culture conditions: 37 LB, aerobic
Expression host: mammalian cells
Induction method: no induction, transient expression
Primers for 5'sequencing: pEGFP-C-5:CATGGTCCTGCTGGAGTTCGTG
Primers for 3'sequencing: primers were designed according to the sequence


pRRLSIN.CPPT.PGK-GFP.WPRE plasmid sequence

LOCUS       Exported                8931 bp ds-DNA     circular SYN 23-JUN-2017
DEFINITION  synthetic circular DNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8931)
  TITLE     Direct Submission
  JOURNAL   Exported Friday, June 23, 2017 from SnapGene Viewer 3.3.4
FEATURES             Location/Qualifiers
     source          1..8931
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..227
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             228..408
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    455..580
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1073..1306
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
     misc_feature    3345..3462
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        3508..4018
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             4040..4759
                     /product="enhanced GFP"
                     /note="mammalian codon-optimized"
     misc_feature    4775..5363
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5246..5257)
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
     LTR             5450..5683
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
     polyA_signal    5755..5876
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      5916..6051
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(6072..6090)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6100..6116)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
     rep_origin      6258..6713
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6739..6843
                     /label=AmpR promoter
     CDS             6844..7704
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
     rep_origin      7875..8463
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
     protein_bind    8751..8772
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8787..8817
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8825..8841
                     /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)."
     primer_bind     8849..8865
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
     promoter        8886..8904
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
        1 tgtagtctta tgcaatactc ttgtagtctt gcaacatggt aacgatgagt tagcaacatg
       61 ccttacaagg agagaaaaag caccgtgcat gccgattggt ggaagtaagg tggtacgatc
      121 gtgccttatt aggaaggcaa cagacgggtc tgacatggat tggacgaacc actgaattgc
      181 cgcattgcag agatattgta tttaagtgcc tagctcgata cataaacggg tctctctggt
      241 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc

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