pKD3 Plasmid

$216.00

  • Model: PVT6001
  • 50 Units in Stock
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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
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