载体基本信息 | |||
载体名称 | pCX-OKS-2A | 载体抗性 | Ampicillin |
---|---|---|---|
载体长度 | 8495 bp | 载体类型 | Mammalian Expression |
拷贝数 | High Copy | 克隆方法 | Restriction Enzyme |
5'引物 | GCGAGCCGCAGCCATTGCCTTTTA | 3'引物 | TTAGCCAGAAGTCAGATGCTC |
pCX-OKS-2A 载体载体图谱
pCX-OKS-2A 载体载体序列
LOCUS Exported 8495 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Non-integrating (episomal) expression of mouse Oct3/4, Klf4 and Sox2. ACCESSION . VERSION . KEYWORDS pCX-OKS-2A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8495) AUTHORS Okita K, Nakagawa M, Hyenjong H, Ichisaka T, Yamanaka S TITLE Generation of Mouse Induced Pluripotent Stem Cells Without Viral Vectors. JOURNAL Science. 2008 Oct 9. ():. PUBMED 18845712 REFERENCE 2 (bases 1 to 8495) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science. 2008 Oct 9. ():." FEATURES Location/Qualifiers source 1..8495 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 4..383 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 385..661 /label=chicken beta-actin promoter intron 662..1678 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1602..1624 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1686..1705 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 2842..2907 /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 4357..4422 /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" primer_bind complement(5516..5535) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 5581..5636 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(5635..5654) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(5997..6013) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5997..6013) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6010..6032) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6021..6037 /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(6045..6075) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6090..6111 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6170..6365 /label=SV40 promoter /note="SV40 early promoter" rep_origin 6216..6351 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6278..6297 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 6371..6505 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6408..6427) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6462..6481 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6573..6590) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6744..7332) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6824..6843) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7503..8363) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 8126..8145 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8364..8468) /gene="bla" /label=AmpR promoter
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