Basic Vector Information
Ef1a_Large T-antigen_Ires_Puro vector Vector Map
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
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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