| 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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