pcDNA5/FRT vector is a 5.1 kb expression vector designed for use with the Flp-In™ System. This vector features: 1. CMV promoter for high level expression in mammalian cells; 2. ten unique restriction site for easy cloning; 3. FLP Recombination Target (FRT) site for Flp recombinase-mediated integration of the vector into the Flp-In™ host cell line and 4. Hygromycin resistance gene for selection of stable cell lines.
| 载体基本信息 | |||
| 载体名称 | pcDNA5/FRT | 载体抗性 | Ampicillin |
|---|---|---|---|
| 载体长度 | 5070 bp | 载体类型 | Mammalian Expression Vectors |
| 载体来源 | Invitrogen | 筛选标记 | Hygromycin |
| 拷贝数 | High copy number | 启动子 | CMV |
| 克隆方法 | Enzyme Cut | 5引物 | T7 promoter |
| 3引物 | BGH Reverse primer | 表达方法 | Constiutive, Stable / Transient |
pcDNA5/FRT 载体载体图谱
pcDNA5/FRT 载体载体序列
LOCUS Exported 5070 bp ds-DNA circular SYN 20-AUG-2020
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA5-FRT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5070)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..5070
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 235..614
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 1028..1252
/note="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
protein_bind 1536..1583
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FRT"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 1591..2613
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/note="HygR"
/note="confers resistance to hygromycin"
/translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
"
polyA_signal 2743..2864
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(2913..2929)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 2937..2953
/bound_moiety="lac repressor encoded by lacI"
/note="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(2961..2991)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 3006..3027
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(3315..3903)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4074..4934)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(4935..5039)
/gene="bla"
/note="AmpR promoter"
