| 质粒名称: | pKOdeltagalK (Kan) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 110089 |
| 存储实验室: | Markus Seeger |
|
载体骨架基本信息 |
|
| 载体名称: | pKO |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌敲除产生 |
| 筛选标记: | -- |
| 载体抗性: | 卡那霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pKOdeltagalK (Kan) was a gift from Markus Seeger (Addgene plasmid # 110089 ; http://n2t.net/addgene:110089 ; RRID:Addgene_110089) |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
质粒图谱.png)
载体简介
载体序列
LOCUS pKOdeltagalK (Kan) 4235 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Analogous to pKO but without the negative selection gene galK..
ACCESSION .
VERSION .
KEYWORDS pKOdeltagalK (Kan)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4235)
TITLE A uniform cloning platform for mycobacterial genetics and protein
production
REFERENCE 2 (bases 1 to 4235)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4235
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(123..938)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 846..865
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 930..949
/label=pENTR-R
/note="pENTR vectors, reverse primer"
primer_bind 1199..1221
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1213..1230
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1214..1230
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(1474..2062)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1554..1573)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 2207..2652
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 2653..4074
/codon_start=1
/gene="Bacillus subtilis sacB"
/product="secreted levansucrase that renders bacterial
growth sensitive to sucrose"
/label=SacB
/note="negative selection marker"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYKVPEFDSSTIKNISSAKGLDVWDSWPLQNTDGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
ORIGIN
1 gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa gttcgattta ttcaacaaag
61 ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct
121 gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca
181 ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc
241 cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa
301 cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg
361 actgaatccg gtgagaatgg caaaagatta tgcatttctt tccagacttg ttcaacaggc
421 cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat
481 tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc
541 gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga
601 tattcttcta atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca
661 tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag
721 tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga
781 aacaactctg gcgcatgggg cttcccatac aagcgataga ttgtcgcacc tgattgcccg
841 acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc
901 ggcctcgagc aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt
961 atgtaagcag acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat
1021 cagagatttt gagacacaac gtggctttcc ccccccccga cgtcaggtgg ccccattcgc
1081 cattcaggct gcgcaactgt tgggaagggc gtttcggtgc gggcctcttc gctattacgc
1141 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
1201 cagtcacgac gttgtaaaac gacggccagt gagcgcgacg taataagttg aagagcatct
1261 gctcttcagc atcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaac
1321 atggtcatag ctgtttcctt gcgtattggg cgctctccgc ttcctcgctc actgactcgc
1381 tgcgctcggt cgttcgggta aagcctgggg tgcctaatga gcaaaaggcc agcaaaaggc
1441 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
1501 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
1561 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
1621 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
1681 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
1741 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
1801 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
1861 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
1921 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
1981 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
2041 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
2101 gtggaacgaa ggcgcgcctg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
2161 cagcgagtca gtgagcgagg aagcggaaga tcgggcccag ccggcccaca tatacctgcc
2221 gttcactatt atttagtgaa atgagatatt atgatatttt ctgaattgtg attaaaaagg
2281 caactttatg cccatgcaac agaaactata aaaaatacag agaatgaaaa gaaacagata
2341 gattttttag ttctttaggc ccgtagtctg caaatccttt tatgattttc tatcaaacaa
2401 aagaggaaaa tagaccagtt gcaatccaaa cgagagtcta atagaatgag gtcgaaaagt
2461 aaatcgcgcg ggtttgttac tgataaagca ggcaagacct aaaatgtgta aagggcaaag
2521 tgtatacttt ggcgtcaccc cttacatatt ttaggtcttt ttttattgtg cgtaactaac
2581 ttgccatctt caaacaggag ggctggaaga agcagaccgc taacacagta cataaaaaag
2641 gagacatgaa cgatgaacat caaaaagttt gcaaaacaag caacagtatt aacctttact
2701 accgcactgc tggcaggagg cgcaactcaa gcgtttgcga aagaaacgaa ccaaaagcca
2761 tataaggaaa catacggcat ttcccatatt acacgccatg atatgctgca aatccctgaa
2821 cagcaaaaaa atgaaaaata taaagttcct gagttcgatt cgtccacaat taaaaatatc
2881 tcttctgcaa aaggcctgga cgtttgggac agctggccat tacaaaacac tgacggcact
2941 gtcgcaaact atcacggcta ccacatcgtc tttgcattag ccggagatcc taaaaatgcg
3001 gatgacacat cgatttacat gttctatcaa aaagtcggcg aaacttctat tgacagctgg
3061 aaaaacgctg gccgcgtctt taaagacagc gacaaattcg atgcaaatga ttctatccta
3121 aaagaccaaa cacaagaatg gtcaggttca gccacattta catctgacgg aaaaatccgt
3181 ttattctaca ctgatttctc cggtaaacat tacggcaaac aaacactgac aactgcacaa
3241 gttaacgtat cagcatcaga cagctctttg aacatcaacg gtgtagagga ttataaatca
3301 atctttgacg gtgacggaaa aacgtatcaa aatgtacagc agttcatcga tgaaggcaac
3361 tacagctcag gcgacaacca tacgctgaga gatcctcact acgtagaaga taaaggccac
3421 aaatacttag tatttgaagc aaacactgga actgaagatg gctaccaagg cgaagaatct
3481 ttatttaaca aagcatacta tggcaaaagc acatcattct tccgtcaaga aagtcaaaaa
3541 cttctgcaaa gcgataaaaa acgcacggct gagttagcaa acggcgctct cggtatgatt
3601 gagctaaacg atgattacac actgaaaaaa gtgatgaaac cgctgattgc atctaacaca
3661 gtaacagatg aaattgaacg cgcgaacgtc tttaaaatga acggcaaatg gtacctgttc
3721 actgactccc gcggatcaaa aatgacgatt gacggcatta cgtctaacga tatttacatg
3781 cttggttatg tttctaattc tttaactggc ccatacaagc cgctgaacaa aactggcctt
3841 gtgttaaaaa tggatcttga tcctaacgat gtaaccttta cttactcaca cttcgctgta
3901 cctcaagcga aaggaaacaa tgtcgtgatt acaagctata tgacaaacag aggattctac
3961 gcagacaaac aatcaacgtt tgcgcctagc ttcctgctga acatcaaagg caagaaaaca
4021 tctgttgtca aagacagcat ccttgaacaa ggacaattaa cagttaacaa ataaaaacgc
4081 aaaagaaaat gccgatatcc tattggcatt gcggccgcgg tctcggcggt gggtcgtcta
4141 gagaaagtga gggagccacg gttggtgaga gctttgttgt aggtggacca gttggtgatt
4201 ttgaactttt gctttgccac ggaatggtct gcgtt
//
