pLV2-CMV-3×FLAG-MCS1-Twinstrep-MCS2-AcGFP1-Puro vector (V014140)

Basic Vector Information

Vector Name:
pLV2-CMV-3×FLAG-MCS1-Twinstrep-MCS2-AcGFP1-Puro
Antibiotic Resistance:
Ampicillin
Length:
8979 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Selection Marker:
Puro
Promoter:
mPGK
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pLV2-CMV-3×FLAG-MCS1-Twinstrep-MCS2-AcGFP1-Puro vector Vector Map

pLV2-CMV-3×FLAG-MCS1-Twinstrep-MCS2-AcGFP1-Puro8979 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088003' LTRHIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSCMV enhancerCMV promoterFLAGStrep-Tag IIStrep-Tag IIAcGFP1PGK promoterPuroRWPRE5' LTRM13 revlac operatorlac promoterCAP binding siteoriAmpRAmpR promoterSV40 poly(A) signal

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 ℃.

pLV2-CMV-3×FLAG-MCS1-Twinstrep-MCS2-AcGFP1-Puro vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V014140                 8979 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014140
VERSION     V014140
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8979)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1303..1536
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1721..1765
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             1914..1955
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2027..2144
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        2201..2504
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2505..2708
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2872..2895
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             2920..2943
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
     CDS             2980..3003
                     /label="Strep-Tag II"
                     /note="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
     CDS             3067..3783
                     /label="AcGFP1"
                     /note="Aequorea coerulescens GFP"
     promoter        3814..4313
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             4334..4930
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    4947..5535
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5742..6375
                     /label="5' LTR"
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6504..6520)
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    complement(6528..6544)
                     /label="lac operator"
                     /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(6552..6582)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6597..6618)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6906..7494)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(7668..8525)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8526..8630)
                     /label="AmpR promoter"
     polyA_signal    8678..8812
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"

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