pKD3
Search name
pKD3,Plasmid pKD3,pKD3 vector
pKD3 information
Prokaryotic resistance: ampicillin Amp, chloramphenicol Chl
Cloned strain: Escherichia coli DH5 alpha
Culture conditions: 37 degrees centigrade
Expression host: Escherichia coli
Use:Homologous recombination HR plasmid
pKD3 description
PKD3 is a gene knockout plasmid for the Red system of Escherichia coli.
pKD3 References
Datsenko, KA, BL Wanner 2000. One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Proc. Natl. Acad. Sci. U.S.A. 97(12):6640-5.
pKD3 sequence
LOCUS Exported 2806 bp ds-DNA circular SYN 27-JAN-2018
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..2806
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind complement(51..98)
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS complement(115..774)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=Chl
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
promoter complement(775..877)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
protein_bind complement(983..1016)
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
rep_origin 1065..1421
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS complement(1518..2378)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=Amp
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(2379..2483)
/gene="bla"
/label=AmpR promoter
terminator 2574..2660
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2752..2779
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
ORIGIN
1 agattgcagc attacacgtc ttgagcgatt gtgtaggctg gagctgcttc gaagttccta
61 tactttctag agaataggaa cttcggaata ggaacttcat ttaaatggcg cgccttacgc
121 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
181 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
241 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
301 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
361 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
421 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
481 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
541 tttcattgcc atacgtaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
601 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
661 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
721 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
781 ttccttagct cctgaaaatc tcgacaactc aaaaaatacg cccggtagtg atcttatttc
841 attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt
901 ggcccagggc ttcccggtat caacagggac accaggattt atttattctg cgaagtgatc
961 ttccgtcaca ggtaggcgcg ccgaagttcc tatactttct agagaatagg aacttcggaa
1021 taggaactaa ggaggatatt catatggacc atggctaatt cccatgtcag ccgttaagtg
1081 ttcctgtgtc actgaaaatt gctttgagag gctctaaggg cttctcagtg cgttacatcc
1141 ctggcttgtt gtccacaacc gttaaacctt aaaagcttta aaagccttat atattctttt
1201 ttttcttata aaacttaaaa ccttagaggc tatttaagtt gctgatttat attaatttta
1261 ttgttcaaac atgagagctt agtacgtgaa acatgagagc ttagtacgtt agccatgaga
1321 gcttagtacg ttagccatga gggtttagtt cgttaaacat gagagcttag tacgttaaac
1381 atgagagctt agtacgtgaa acatgagagc ttagtacgta ctatcaacag gttgaactgc
1441 ggatcttgcg gccgcaaaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
1501 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
1561 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
1621 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
1681 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
1741 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
1801 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
1861 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
1921 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
1981 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
2041 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
2101 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
2161 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
2221 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
2281 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
2341 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
2401 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
文献和实验
技术资料
