- 载体名称:
- pcDNA3.1-ccdB-3xFLAG-V5
- 载体抗性:
- Chloramphenicol, Ampicillin
- 载体长度:
- 7412 bp
- 载体类型:
- Mammalian Expression
- 复制子:
- ori
- 筛选标记:
- Hygromycin
- 拷贝数:
- High Copy
pcDNA3.1-ccdB-3xFLAG-V5 载体载体图谱
pcDNA3.1-ccdB-3xFLAG-V5 载体载体序列
LOCUS 40924_10101 7412 bp DNA circular SYN 13-MAY-2021 DEFINITION Mammalian Gateway-compatible expression vector backbone with a C-terminal 3xFLAG-V5 tag. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7412) AUTHORS Taipale M, Krykbaeva I, Koeva M, Kayatekin C, Westover KD, Karras GI, Lindquist S TITLE Quantitative analysis of HSP90-client interactions reveals principles of substrate recognition. JOURNAL Cell. 2012 Aug 31;150(5):987-1001. doi: 10.1016/j.cell.2012.06.047. PUBMED 22939624 REFERENCE 2 (bases 1 to 7412) TITLE Direct Submission REFERENCE 3 (bases 1 to 7412) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1016/j.cell.2012.06"; journalName: "Cell"; date: "2012-08-31- 31"; volume: "150"; issue: "5"; pages: "987-1001" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..7412 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 969..1093 /label=attR1 /note="recombination site for the Gateway(R) LR reaction" promoter 1118..1148 /label=lac UV5 promoter /note="E. coli lac promoter with an 'up' mutation" CDS 1202..1858 /codon_start=1 /label=CmR /note="chloramphenicol acetyltransferase" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" CDS 2203..2505 /codon_start=1 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" protein_bind complement(2549..2673) /label=attR2 /note="recombination site for the Gateway(R) LR reaction" CDS 2675..2740 /codon_start=1 /label=3xFLAG /note="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 2759..2800 /codon_start=1 /label=V5 tag /note="epitope tag from simian virus 5" /translation="GKPIPNPLLGLDST" polyA_signal 2850..3064 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3110..3538 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 3552..3881 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 3930..4952 /codon_start=1 /label=HygR /note="aminoglycoside phosphotransferase from E. coli" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" polyA_signal 5085..5218 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5255..5271) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(5279..5295) /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(5303..5333) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(5348..5369) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(5486..5503) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5657..6245) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6419..7276) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(7277..7381) /label=AmpR promoter
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