Basic Vector Information
OKSIM 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 ℃.
OKSIM vector Sequence
LOCUS OKSIM. 12417 bp DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA. ACCESSION . VERSION . KEYWORDS OKSIM. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12417) AUTHORS Ross PJ, Suhr S, Rodriguez RM, Chang EA, Wang K, Siripattarapravat K, Ko T, Cibelli JB TITLE Human Induced Pluripotent Stem Cells Produced Under Xeno-Free Conditions. JOURNAL Stem Cells Dev. 2009 Dec 23. ():. PUBMED 20030562 REFERENCE 2 (bases 1 to 12417) TITLE Direct Submission REFERENCE 3 (bases 1 to 12417) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Stem Cells Dev. 2009 Dec 23. ():." COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..12417 /mol_type="other DNA" /organism="synthetic DNA construct" misc_feature 13..563 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 608..1924 /label=c-Myc /note="human c-Myc proto-oncogene" primer_bind complement(1978..1994) /label=KS primer /note="common sequencing primer, one of multiple similar variants" misc_feature 2008..2596 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" protein_bind complement(2740..2773) /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." LTR 2793..2973 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" polyA_signal 3002..3226 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" regulatory 3269..3278 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" promoter 3322..3426 /label=AmpR promoter CDS 3427..4284 /label=AmpR /note="beta-lactamase" rep_origin 4458..5046 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" LTR 5205..5838 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 5885..6010 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 6503..6736 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 6921..6965 /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 7114..7155 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" misc_feature 7252..7369 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 7498..8676 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" promoter 8692..8710 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" CDS 8736..9815 /label=hOct4 /note="Homo sapiens Oct-4 gene. Encodes a transcription factor containing a POU homeodomain that plays a key role in embryonic development and stem cell pluripotency. Aberrant expression in adult tissues is associated with tumorigenesis." CDS 9840..9905 /codon_start=1 /product="2A peptide from foot-and-mouth disease virus polyprotein" /label=F2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="VKQTLNFDLLKLAGDVESNPGP" misc_feature 10000..10576 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 11352..11417 /codon_start=1 /product="2A peptide from foot-and-mouth disease virus polyprotein" /label=F2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="VKQTLNFDLLKLAGDVESNPGP" CDS 11430..12380 /label=hSOX2 /note="Homo sapiens transcription factor SOX-2 gene. Belongs to the SRY-related HMG-box (SOX) family of transcription factors, which is involved in the regulation of embryonic development and in the determination of cell fate"
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