pKD13 contains R6K gamma ori, which requires λ pir protein for its initiation. pKD13 therefore cannot replicate in conventional strains such as DH5α and can only proliferate in bacteria that contain pir+ genes (which can express λ pir protein), such as S17-1λpir or DH5αλpir.
| 载体基本信息 | |||
| 载体名称 | pKD13 | 载体抗性 | Ampicillin, Kanamycin |
|---|---|---|---|
| 载体长度 | 3434 bp | 载体类型 | Knockout Vectors |
| 克隆方法 | Enzyme digestion and ligation | ||
pKD13 载体载体图谱
pKD13 载体载体序列
LOCUS Exported 3434 bp ds-DNA circular SYN 13-JAN-2022
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3434)
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..3434
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind complement(51..98)
/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 complement(111..905)
/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(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
protein_bind complement(1273..1306)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FRT (minimal)"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
terminator 1386..1632
/note="lambda tL3 terminator"
/note="transcription terminator tL3 from phage lambda"
rep_origin 1693..2049
/note="R6K gamma ori"
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS complement(2146..3006)
/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(3007..3111)
/gene="bla"
/note="AmpR promoter"
terminator 3202..3288
/gene="Escherichia coli rrnB"
/note="rrnB T1 terminator"
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 3380..3407
/note="rrnB T2 terminator"
/note="transcription terminator T2 from the E. coli rrnB
gene"
