| 出品公司: | Barry L. Wanner实验室 |
|---|---|
| 载体名称: | pKD3 |
| 质粒类型: | 细菌表达；基因敲除系统 |
| 高拷贝/低拷贝: | 高拷贝 |
| 启动子: | R6K |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 2842 bp |
| 5' 测序引物及序列: | R6K-3' |
| 3' 测序引物及序列: | rrnB-T1-term-F |
| 载体标签: | -- |
| 载体抗性: | 氨苄和氯霉素 |
| 备注: | 用于细菌基因敲除 |
| Addgene: | 45604 |
| 稳定性: | 稳定 |
| 组成型: | 非组成 |
| 病毒/非病毒: | 非病毒 |
| 参考文献： | One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. Datsenko KA, Wanner BL. Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6640-5. 10.1073/pnas.120163297 |
买家导航
pKD3载体质粒基本信息
pKD3质粒图谱载体图谱和pKD3载体序列质粒序列多克隆位点信息
pKD3质粒载体简介
pKD3质粒序列载体序列
LOCUS pKD3 2842 bp ds-DNA circular SYN 28-APR-2020
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pKD3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 2842)
AUTHORS Datsenko KA, Wanner BL
TITLE One-step inactivation of chromosomal genes in Escherichia coli K-12
using PCR products.
JOURNAL Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6640-5.
PUBMED 10829079
REFERENCE 2 (bases 1 to 2842)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 28, 2020 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..2842
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 107..193
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 285..312
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
protein_bind complement(390..437)
/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(454..1113)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind 1028..1047
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
promoter complement(1114..1216)
/label=cat promoter
/note="promoter of the E. coli cat gene encoding
chloramphenicol acetyltransferase"
protein_bind complement(1322..1355)
/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 1404..1792
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
CDS complement(1894..2754)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 2517..2536
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2755..17)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcatcgat ggcccccgat
61 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaatgaaa
121 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
181 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg
241 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac
301 ggatggcctt tttgcgtggc cagtgccaag cttgcatgca gattgcagca ttacacgtct
361 tgagcgattg tgtaggctgg agctgcttcg aagttcctat actttctaga gaataggaac
421 ttcggaatag gaacttcatt taaatggcgc gccttacgcc ccgccctgcc actcatcgca
481 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg
541 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt
601 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact
661 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc
721 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc
781 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta
841 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacgtaattc
901 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt
961 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt
1021 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc
1081 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct
1141 cgacaactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc
1201 tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc
1261 aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag gtaggcgcgc
1321 cgaagttcct atactttcta gagaatagga acttcggaat aggaactaag gaggatattc
1381 atatggacca tggctaattc ccatgtcagc cgttaagtgt tcctgtgtca ctcaaaattg
1441 ctttgagagg ctctaagggc ttctcagtgc gttacatccc tggcttgttg tccacaaccg
1501 ttaaacctta aaagctttaa aagccttata tattcttttt tttcttataa aacttaaaac
1561 cttagaggct atttaagttg ctgatttata ttaattttat tgttcaaaca tgagagctta
1621 gtacgtgaaa catgagagct tagtacgtta gccatgagag cttagtacgt tagccatgag
1681 ggtttagttc gttaaacatg agagcttagt acgttaaaca tgagagctta gtacgtgaaa
1741 catgagagct tagtacgtac tatcaacagg ttgaactgct gatcttcaga tcctctacgc
1801 cggacgcatc gtggccggat cttgcggccg caaaaattaa aaatgaagtt ttaaatcaat
1861 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
1921 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
1981 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
2041 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
2101 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
2161 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
2221 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
2281 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
2341 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
2401 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
2461 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
2521 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
2581 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
2641 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
2701 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
2761 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
2821 tgaatgtatt tagaaaaata aa
//
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