载体基本信息 | |||
载体名称 | p3xFLAG-CMV-14 | 载体抗性 | Ampicillin |
---|---|---|---|
载体长度 | 6310 bp | 载体类型 | Mammalian Expression Vectors |
载体来源 | Sigma-Aldrich | 筛选标记 | Neomycin/G418(Geneticin) |
拷贝数 | High copy number | 启动子 | CMV |
克隆方法 | Enzyme digestion and ligation | 5'引物 | M13 fwd |
3'引物 | M13 rev |
p3xFLAG-CMV-14 载体载体图谱
p3xFLAG-CMV-14 载体载体序列
LOCUS Exported 6310 bp ds-DNA circular SYN 28-11-2013 DEFINITION Vector with a neomycin resistance marker for expression of C-terminally 3xFLAG-tagged proteins in mammalian cells. ACCESSION . VERSION . KEYWORDS p3xFLAG-CMV-14 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6310) AUTHORS Sigma-Aldrich TITLE Direct Submission FEATURES Location/Qualifiers source 1..6310 /organism="synthetic DNA construct" /lab_host="Mammalian Cells" /mol_type="other DNA" primer_bind 141..157 /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" enhancer 318..697 /note="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 698..901 /note="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" misc_feature 924..990 /note="MCS" /note="multiple cloning site" CDS 993..1058 /codon_start=1 /product="three tandem FLAG epitope tags, followed by an enterokinase cleavage site" /note="3xFLAG" /translation="DYKDHDGDYKDHDIDYKDDDDK" polyA_signal 1069..1691 /note="hGH poly(A) signal" /note="human growth hormone polyadenylation signal" promoter 1733..2030 /note="SV40 promoter" /note="SV40 enhancer and early promoter" CDS 2084..2878 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /note="NeoR/KanR" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin)" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" polyA_signal 3530..3664 /note="SV40 poly(A) signal" /note="SV40 polyadenylation signal" primer_bind complement(3702..3718) /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" rep_origin complement(4104..4692) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4863..5723) /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(5724..5828) /gene="bla" /note="AmpR promoter" rep_origin 5855..6310 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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