载体基本信息 | |||
载体名称 | pcDNA-DEST53 | 载体抗性 | Ampicillin |
---|---|---|---|
载体长度 | 7767 bp | 载体类型 | Gateway Cloning Vectors |
载体来源 | Invitrogen (Life Technologies) | 筛选标记 | Neomycin/G418(Geneticin) |
拷贝数 | High copy number |
pcDNA-DEST53 载体载体图谱
pcDNA-DEST53 载体载体序列
LOCUS 40924_9836 7767 bp DNA circular SYN 01-JAN-1980 DEFINITION Gateway(R) destination vector for expressing N-terminally GFP-tagged proteins in mammalian cells. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7767) AUTHORS Invitrogen (Life Technologies) TITLE Direct Submission REFERENCE 2 (bases 1 to 7767) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" FEATURES Location/Qualifiers source 1..7767 /mol_type="other DNA" /organism="synthetic DNA construct" 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" CDS 905..1621 /codon_start=1 /label=Cycle 3 GFP /note="Cycle 3 GFP (Crameri et al., 1996)" /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITHGMDELYK" protein_bind 1643..1767 /label=attR1 /note="recombination site for the Gateway(R) LR reaction" promoter 1792..1822 /label=lac UV5 promoter /note="E. coli lac promoter with an 'up' mutation" CDS 1876..2532 /codon_start=1 /label=CmR /note="chloramphenicol acetyltransferase" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" CDS 2856..3158 /codon_start=1 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" protein_bind complement(3202..3326) /label=attR2 /note="recombination site for the Gateway(R) LR reaction" polyA_signal 3364..3588 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 3634..4062 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 4076..4405 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 4472..5263 /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 5440..5573 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5610..5626) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind complement(5634..5650) /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(5658..5688) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind complement(5703..5724) /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(6012..6600) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6774..7631) /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(7632..7736) /label=AmpR promoter
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