Ef1a_Large T-antigen_Ires_Puro vector (V006660)

Basic Vector Information

      • Vector Name:
      • Ef1a_Large T-antigen_Ires_Puro
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 11672 bp
      • Type:
      • Mammalian Expression, Lentiviral, Cre/Lox
      • Replication origin:
      • ori
      • Selection Marker:
      • Puromycin
      • Promoter:
      • EF-1α
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • Ef1a

Ef1a_Large T-antigen_Ires_Puro vector Vector Map

Ef1a_Large T-antigen_Ires_Puro11672 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000105001100011500large T antigenIRES2PuroRloxP5' LTR (truncated)bGH poly(A) signalf1 oriSV40 promoterEM7 promoterBleoRSV40 poly(A) signalM13 revlac operatorlac promoterCAP binding siteL4440oriAmpRAmpR promoterpRS-markerCMV enhancerCMV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509cPPT/CTSEF-1-alpha promoterT7 promoter

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 ℃.

Ef1a_Large T-antigen_Ires_Puro vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Ef1a_Large_T-ant       11672 bp DNA     circular SYN 16-AUG-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    Ef1a_Large T-antigen_Ires_Puro.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11672)
  AUTHORS   Mali P, Ye Z, Hommond HH, Yu X, Lin J, Chen G, Zou J, Cheng L
  TITLE     Improved efficiency and pace of generating induced pluripotent stem 
            cells from human adult and fetal fibroblasts.
  JOURNAL   Stem Cells. 2008 Aug . 26(8):1998-2005.
  PUBMED    18511599
REFERENCE   2  (bases 1 to 11672)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 11672)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cells.
            2008 Aug . 26(8):1998-2005."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11672
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..2124
                     /label=large T antigen
                     /note="large T antigen from SV40 (simian virus 40)"
     misc_feature    2159..2745
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2746..3342
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     protein_bind    complement(3459..3492)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     LTR             3517..3697
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    3729..3953
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3999..4427
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4441..4770
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        4818..4865
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             4884..5255
                     /label=BleoR
                     /note="antibiotic-binding protein"
     polyA_signal    5388..5521
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5558..5574)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5558..5574)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5571..5593)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5582..5598
                     /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(5606..5636)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5651..5672)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(5789..5806)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5960..6548)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6722..7579)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7580..7684)
                     /label=AmpR promoter
     primer_bind     complement(7759..7778)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     enhancer        7950..8329
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8330..8532
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             8547..8727
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    8774..8899
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    9392..9625
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             9810..9854
                     /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             10003..10044
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    10152..10269
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        10405..11583
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     promoter        11600..11618
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"

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