Cre-IRES-PuroR vector (V001194)

Basic Vector Information

      • Vector Name:
      • Cre-IRES-PuroR
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 9229 bp
      • Type:
      • Mammalian Expression, Lentiviral
      • Selection Marker:
      • Puromycin
      • Copy Number:
      • High Copy
      • Promoter:
      • EF1a
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • GGTTCATTCTCAAGCCTCAGACAGTG

Cre-IRES-PuroR vector Vector Map

Cre-IRES-PuroR9229 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088009200EBV-rev3' LTRHIV-1 PsiRREgp41 peptideProtein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509cPPT/CTSEF-1-alpha promoterKozak sequenceCreIRES2PuroRWPRE5' LTR (truncated)pBRforEcoAmpR promoterAmpRoriL4440SV40pro-FCAP binding sitelac promoterlac operator

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 ℃.

Cre-IRES-PuroR vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Cre-IRES-PuroR.        9229 bp DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    Cre-IRES-PuroR.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9229)
  AUTHORS   Somers A, Jean JC, Sommer CA, Omari A, Ford CC, Mills JA, Ying L, 
            Sommer AG, Jean JM, Smith BW, Lafyatis R, Demierre MF, Weiss DJ, 
            French DL, Gadue P, Murphy GJ, Mostoslavsky G, Kotton DN
  TITLE     Generation of transgene-free lung disease-specific human induced 
            pluripotent stem cells using a single excisable lentiviral stem cell
            cassette.
  JOURNAL   Stem Cells. 2010 Oct . 28(10):1728-40.
  PUBMED    20715179
REFERENCE   2  (bases 1 to 9229)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9229)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cells.
            2010 Oct . 28(10):1728-40."
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9229
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..74
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     LTR             266..899
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    946..1071
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1568..1801
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1986..2030
                     /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             2179..2220
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2323..2440
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2488..3666
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     regulatory      3671..3680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3707..4735
                     /label=Cre
                     /note="site-specific recombinase"
     misc_feature    4741..5319
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             5323..5919
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    5936..6524
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6672..6852
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6899..6917)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        6985..7089
                     /label=AmpR promoter
     CDS             7090..7947
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8121..8709
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8863..8880
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     8972..8991
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     protein_bind    9120..9141
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9156..9186
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9194..9210
                     /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)."

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