pCEP4 vector is designed for high-level, constitutive expression from either the CMV or RSV promoters. Both vectors contain the EBNA-1 gene for episomal expression in primate and canine cell lines.
| 载体基本信息 | |||
| 载体名称 | pCEP4 | 载体长度 | 10188 bp |
|---|---|---|---|
| 载体类型 | Mammalian Expression Vectors | 载体来源 | Thermo Fisher Scientific |
| 筛选标记 | Hygromycin | 拷贝数 | High copy number |
| 启动子 | CMV | 克隆方法 | Enzyme Cut |
| 表达方法 | Constiutive, Stable / Transient | ||
pCEP4 载体载体图谱
pCEP4 载体载体序列
LOCUS Exported 10188 bp ds-DNA circular SYN 28-JAN-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCEP4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10188)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..10188
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 384..587
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
polyA_signal 672..806
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 1531..3320
/note="oriP"
/note="Epstein-Barr virus oriP replication origin (Yates et
al., 2000)"
CDS complement(3622..5547)
/codon_start=1
/note="EBNA-1"
/note="EBNA1 activates the initiation of viral DNA
replication through binding to specific sites in the viral
latent origin of replication, oriP. Additionally, it
governs the segregation of viral episomes by mediating
their attachment to host cell metaphase chromosomes. Also
activates the transcription of several viral latency genes.
Finally, it can counteract the stabilization of host
p53/TP53 by host USP7, thereby decreasing apoptosis and
increasing host cell survival."
/translation="MSDEGPGTGPGNGLGEKGDTSGPEGSGGSGPQRRGGDNHGRGRGR
GRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGTHGGTGAGAGAGGAGAGGAG
AGGGAGAGGGAGGAGGAGGAGAGGGAGAGGGAGGAGGAGAGGGAGAGGGAGGAGAGGGA
GGAGGAGAGGGAGAGGGAGGAGAGGGAGGAGGAGAGGGAGAGGAGGAGGAGAGGAGAGG
GAGGAGGAGAGGAGAGGAGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGGAGAG
GGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRG
GSGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPP
PGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGR
RKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSL
YNLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLK
DAIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDE
GEEGQE"
promoter 6068..6172
/gene="bla"
/note="AmpR promoter"
CDS 6173..7033
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 7204..7792
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 8223..8368
/note="HSV TK promoter"
/note="herpes simplex virus thymidine kinase promoter"
CDS 8411..9448
/codon_start=1
/product="hygromycin B phosphotransferase"
/note="HygR"
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPDRE
MGEAN"
polyA_signal 9490..9538
/note="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
