Basic Vector Information
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.
Basic Vector Information | |||
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Vector Name | pcDNA5/FRT | Antibiotic Resistance | Ampicillin |
Length | 5070 bp | Type | Mammalian Expression Vectors |
Source | Invitrogen | Selection Marker | Hygromycin |
Copy Number | High copy number | Promoter | CMV |
Cloning Method | Enzyme Cut | 5' Primer | T7 promoter |
3' Primer | BGH Reverse primer | Expression Method | Constiutive, Stable / Transient |
pcDNA5/FRT vector Vector Map
pcDNA5/FRT vector Sequence
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) AUTHORS caoheibi TITLE Direct Submission JOURNAL Exported Thursday, August 20, 2020 from SnapGene 3.2.1 http://www.snapgene.com 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"
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