- 载体名称:
- pCX-OKS-2A
- 载体抗性:
- Ampicillin
- 载体长度:
- 8495 bp
- 载体类型:
- Mammalian Expression
- 复制子:
- ori
- 拷贝数:
- High Copy
- 启动子:
- CAG
- 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 40924_13930 8495 bp DNA circular SYN 13-MAY-2021
DEFINITION Non-integrating (episomal) expression of mouse Oct3/4, Klf4 and
Sox2.
ACCESSION .
VERSION .
KEYWORDS .
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)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 8495)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science.
2008 Oct 9. ():."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..8495
/mol_type="other DNA"
/organism="synthetic DNA construct"
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 1686..1705
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS 1762..2817
/codon_start=1
/label=mPou5f1
/note="Mus musculus Oct-4 gene. Encodes a transcription
factor containing a POU homeodomain that plays a key role
in embryonic development and stem cell pluripotency."
/translation="MAGHLASDFAFSPPPGGGDGSAGLEPGWVDSRTWLSFQGPPGGPG
IGPGSEVLGISPCPPAYEFCGGMAYCGPQVGLGLVPQVGVETLQPEGQAGARVESNSEG
TSSEPCADRPNAVKLEKVEPTPEESQDMKALQKELEQFAKLLKQKRITLGYTQADVGLT
LGVLFGKVFSQTTICRFEALQLSLKNMCKLRPLLEKWVEEADNNENLQEICKSETLVQA
RKRKRTSIENRVRWSLETMFLKCPKPSLQQITHIANQLGLEKDVVRVWFCNRRQKGKRS
SIEYSQREEYEATGTPFPGGAVSFPLPPGPHFGTPGYGSPHFTTLYSVPFPEGEAFPSV
PVTALGSPMHSN"
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 2911..4332
/codon_start=1
/label=mKlf4
/note="Mouse Kruppel-like factor (Klf4) gene. Belongs to
the relatively large family of SP1-like transcription
factors and is involved in the regulation of proliferation,
differentiation, apoptosis and somatic cell reprogramming."
/translation="MAVSDALLPSFSTFASGPAGREKTLRPAGAPTNRWREELSHMKRL
PPLPGRPYDLAATVATDLESGGAGAACSSNNPALLARRETEEFNDLLDLDFILSNSLTH
QESVAATVTTSASASSSSSPASSGPASAPSTCSFSYPIRAGGDPGVAASNTGGGLLYSR
ESAPPPTAPFNLADINDVSPSGGFVAELLRPELDPVYIPPQQPQPPGGGLMGKFVLKAS
LTTPGSEYSSPSVISVSKGSPDGSHPVVVAPYSGGPPRMCPKIKQEAVPSCTVSRSLEA
HLSAGPQLSNGHRPNTHDFPLGRQLPTRTTPTLSPEELLNSRDCHPGLPLPPGFHPHPG
PNYPPFLPDQMQSQVPSLHYQELMPPGSCLPEEPKPKRGRRSWPRKRTATHTCDYAGCG
KTYTKSSHLKAHLRTHTGEKPYHCDWDGCGWKFARSDELTRHYRKHTGHRPFQCQKCDR
AFSRSDHLALHMKRHF"
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"
CDS 4447..5403
/codon_start=1
/label=mSox2
/note="Mus musculus 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"
/translation="MYNMMETELKPPGPQQASGGGGGGGNATAAATGGNQKNSPDRVKR
PMNAFMVWSRGQRRKMAQENPKMHNSEISKRLGAEWKLLSETEKRPFIDEAKRLRALHM
KEHPDYKYRPRRKTKTLMKKDKYTLPGGLLAPGGNSMASGVGVGAGLGAGVNQRMDSYA
HMNGWSNGSYSMMQEQLGYPQHPGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGS
PTYSMSYSQQGTPGMALGSMGSVVKSEASSSPPVVTSSSHSRAPCQAGDLRDMISMYLP
GAEVPEPAAPSRLHMAQHYQSGPVPGTAINGTLPLSHM"
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"
protein_bind complement(6021..6037)
/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)."
promoter complement(6045..6075)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(6090..6111)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6169..6365
/label=SV40 promoter
/note="SV40 early promoter"
polyA_signal 6371..6505
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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"
CDS complement(7506..8363)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8364..8468)
/label=AmpR promoter
promoter 8495
/label=CAG
/note="CMV early enhancer fused to modified chicken
beta-actin promoter"
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