Lentiviral-TOP-dGFP-reporter vector (V012444)

Basic Vector Information

      • Vector Name:
      • Lentiviral-TOP-dGFP-reporter
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7151 bp
      • Type:
      • Mammalian Expression, Lentiviral
      • Copy Number:
      • High Copy
      • Cloning Method:
      • Restriction Enzyme

Lentiviral-TOP-dGFP-reporter vector Vector Map

Lentiviral-TOP-dGFP-reporter7151 bp30060090012001500180021002400270030003300360039004200450048005100540057006000630066006900oriL4440CAP binding sitelac promoterlac operatorM13 revT3RSV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptidecPPT/CTSKozak sequenceKozak sequenceWPRE3' LTR (Delta-U3)SV40 poly(A) signalSV40 oriT7M13 Forwardf1 oriAmpR promoterAmpR

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 ℃.

Lentiviral-TOP-dGFP-reporter vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                7151 bp ds-DNA     circular SYN 17-AUG-2021
DEFINITION  Beta-catenin reporter - 3rd generation self-inactivating lentiviral 
            plasmid LEF-1/TCF-responsive promoter driven d2-eGFP (destabilized, 
            half-life of 2h)..
ACCESSION   .
VERSION     .
KEYWORDS    Lentiviral-TOP-dGFP-reporter
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7151)
  AUTHORS   Reya T, Duncan AW, Ailles L, Domen J, Scherer DC, Willert K, Hintz 
            L, Nusse R, Weissman IL
  TITLE     A role for Wnt signalling in self-renewal of haematopoietic stem 
            cells.
  JOURNAL   Nature. 2003 May 22. 423(6938):409-14.
  PUBMED    12717450
REFERENCE   2  (bases 1 to 7151)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature. 
            2003 May 22. 423(6938):409-14."
FEATURES             Location/Qualifiers
     source          1..7151
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      15..603
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     504..523
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     757..774
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    891..912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        927..957
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    965..981
                     /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     970..992
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     989..1005
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     989..1005
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1024..1044
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        1026..1044
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        1072..1298
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             1299..1479
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1526..1651
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2144..2377
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2562..2606
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2873..2990
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     regulatory      3169..3178
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      3194..3203
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3200..4045
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(3245..3266)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3506..3525)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3853..3874
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4066..4654
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4119..4139)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4537..4548)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             4741..4974
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5046..5167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5083..5102)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5137..5156
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5207..5342
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5269..5288
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(5362..5381)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5363..5381)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5391..5408)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5391..5407)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5400..5422)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5549..6004
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5636..5655)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5846..5867
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6030..6134
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6135..6995
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6353..6372)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"

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