| 质粒名称: | pKO (Hyg) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 110087 |
| 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: | 潮霉素 |
| 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: | pKO (Hyg) was a gift from Markus Seeger (Addgene plasmid # 110087 ; http://n2t.net/addgene:110087 ; RRID:Addgene_110087) |
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 pKO (Hyg) 6203 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Suicide vector for generation of gene deletions in mycobacteria.
Contains the two genes sacB and galK for negative selection. .
ACCESSION .
VERSION .
KEYWORDS pKO (Hyg)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6203)
TITLE A uniform cloning platform for mycobacterial genetics and protein
production
REFERENCE 2 (bases 1 to 6203)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6203
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 21..51
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind 57..76
/label=lac operator (symmetric)
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG). The
symmetric lac operator was optimized for tight binding of
lac repressor."
primer_bind 1652..1674
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1666..1683
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1667..1683
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(1927..2515)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2007..2026)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 2660..3105
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 3106..4527
/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 catggatcca gaccccaggc ttgacacttt atgcttccgg ctcgtataat gtgtggaatt
61 gtgagcgctc acaattcgca tgcggtacca agctgatccg ataacacagg aacagatctt
121 ccggagtgta agaaatgagt ctgaaagaaa aaacacaatc tctgtttgcc aacgcatttg
181 gctaccctgc cactcacacc attcaggcgc ctggccgcgt gaatttgatt ggtgaacaca
241 ccgactacaa cgacggtttc gttctgccct gcgcgattga ttatcaaacc gtgatcagtt
301 gtgcaccacg cgatgaccgt aaagttcgcg tgatggcagc cgattatgaa aatcagctcg
361 acgagttttc cctcgatgcg cccattgtcg cacatgaaaa ctatcaatgg gctaactacg
421 ttcgtggcgt ggtgaaacat ctgcaactgc gtaacaacag cttcggcggc gtggacatgg
481 tgatcagcgg caatgtgccg cagggtgccg ggttaagttc ttccgcttca ctggaagtcg
541 cggtcggaac cgtattgcag cagctttatc atctgccgct ggacggcgca caaatcgcgc
601 ttaacggtca ggaagcagaa aaccagtttg taggctgtaa ctgcgggatc atggatcagc
661 taatttccgc gctcggcaag aaagatcatg ccttgctgat cgattgccgc tcactgggga
721 ccaaagcagt ttccatgccc aaaggtgtgg ctgtcgtcat catcaacagt aacttcaaac
781 gtaccctggt tggcagcgaa tacaacaccc gtcgtgaaca gtgcgaaacc ggtgcgcgtt
841 tcttccagca gccagccctg cgtgatgtca ccattgaaga gttcaacgct gttgcgcatg
901 aactggaccc gatcgtggca aaacgcgtgc gtcatatact gactgaaaac gcccgcaccg
961 ttgaagctgc cagcgcgctg gagcaaggcg acctgaaacg tatgggcgag ttgatggcgg
1021 agtctcatgc ctctatgcgc gatgatttcg aaatcaccgt gccgcaaatt gacactctgg
1081 tagaaatcgt caaagctgtg attggcgaca aaggtggcgt acgcatgacc ggcggcggat
1141 ttggcggctg tatcgtcgcg ctgatcccgg aagaactggt gcctgccgta cagcaagctg
1201 tcgctgaaca atatgaagca aaaacaggta ttaaagagac tttttacgtt tgtaaaccat
1261 cacaaggagc aggacagtgc tgaacgaaac tcctgcagcg cgtagtacct cacaccgtcc
1321 tgcgggttga cgtatgaggc ttggatcctc ccgctgcgct gagtcttcag cgatccccat
1381 ccgcgacgtg ccaactaggt ctcctctcgt cgtgaacaag gctaccgggt tgcaactcct
1441 gtgcaactct caggcttcaa cgcgcttcta cgacctgcaa tttctttcca cttagaggat
1501 gcagccgaga gggggtaaaa acctcccatt cgccattcag gctgcgcaac tgttgggaag
1561 ggcgtttcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
1621 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
1681 agtgagcgcg acgtaataag ttgaagagca tctgctcttc agcatcactg cccgctttcc
1741 agtcgggaaa cctgtcgtgc cagctgcatt aacatggtca tagctgtttc cttgcgtatt
1801 gggcgctctc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg gtaaagcctg
1861 gggtgcctaa tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
1921 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
1981 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
2041 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
2101 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
2161 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
2221 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
2281 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
2341 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
2401 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
2461 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
2521 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaggcgcgc ctgagtgagc
2581 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
2641 agatcgggcc cagccggccc acatatacct gccgttcact attatttagt gaaatgagat
2701 attatgatat tttctgaatt gtgattaaaa aggcaacttt atgcccatgc aacagaaact
2761 ataaaaaata cagagaatga aaagaaacag atagattttt tagttcttta ggcccgtagt
2821 ctgcaaatcc ttttatgatt ttctatcaaa caaaagagga aaatagacca gttgcaatcc
2881 aaacgagagt ctaatagaat gaggtcgaaa agtaaatcgc gcgggtttgt tactgataaa
2941 gcaggcaaga cctaaaatgt gtaaagggca aagtgtatac tttggcgtca ccccttacat
3001 attttaggtc tttttttatt gtgcgtaact aacttgccat cttcaaacag gagggctgga
3061 agaagcagac cgctaacaca gtacataaaa aaggagacat gaacgatgaa catcaaaaag
3121 tttgcaaaac aagcaacagt attaaccttt actaccgcac tgctggcagg aggcgcaact
3181 caagcgtttg cgaaagaaac gaaccaaaag ccatataagg aaacatacgg catttcccat
3241 attacacgcc atgatatgct gcaaatccct gaacagcaaa aaaatgaaaa atataaagtt
3301 cctgagttcg attcgtccac aattaaaaat atctcttctg caaaaggcct ggacgtttgg
3361 gacagctggc cattacaaaa cactgacggc actgtcgcaa actatcacgg ctaccacatc
3421 gtctttgcat tagccggaga tcctaaaaat gcggatgaca catcgattta catgttctat
3481 caaaaagtcg gcgaaacttc tattgacagc tggaaaaacg ctggccgcgt ctttaaagac
3541 agcgacaaat tcgatgcaaa tgattctatc ctaaaagacc aaacacaaga atggtcaggt
3601 tcagccacat ttacatctga cggaaaaatc cgtttattct acactgattt ctccggtaaa
3661 cattacggca aacaaacact gacaactgca caagttaacg tatcagcatc agacagctct
3721 ttgaacatca acggtgtaga ggattataaa tcaatctttg acggtgacgg aaaaacgtat
3781 caaaatgtac agcagttcat cgatgaaggc aactacagct caggcgacaa ccatacgctg
3841 agagatcctc actacgtaga agataaaggc cacaaatact tagtatttga agcaaacact
3901 ggaactgaag atggctacca aggcgaagaa tctttattta acaaagcata ctatggcaaa
3961 agcacatcat tcttccgtca agaaagtcaa aaacttctgc aaagcgataa aaaacgcacg
4021 gctgagttag caaacggcgc tctcggtatg attgagctaa acgatgatta cacactgaaa
4081 aaagtgatga aaccgctgat tgcatctaac acagtaacag atgaaattga acgcgcgaac
4141 gtctttaaaa tgaacggcaa atggtacctg ttcactgact cccgcggatc aaaaatgacg
4201 attgacggca ttacgtctaa cgatatttac atgcttggtt atgtttctaa ttctttaact
4261 ggcccataca agccgctgaa caaaactggc cttgtgttaa aaatggatct tgatcctaac
4321 gatgtaacct ttacttactc acacttcgct gtacctcaag cgaaaggaaa caatgtcgtg
4381 attacaagct atatgacaaa cagaggattc tacgcagaca aacaatcaac gtttgcgcct
4441 agcttcctgc tgaacatcaa aggcaagaaa acatctgttg tcaaagacag catccttgaa
4501 caaggacaat taacagttaa caaataaaaa cgcaaaagaa aatgccgata tcctattggc
4561 attgcggccg cggtctcggc ggtgggtcgt ctagaggtct gacgctcagt ggaacgaaaa
4621 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
4681 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
4741 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
4801 agttgcctga ctccccgacg tggccgacca gcccgtcatc gtcaacgcct gatccgcggt
4861 gcggacaggc cgtgtcgcga ccggccgtgc ggaattaagc cggcccgtac cctgtgaata
4921 gaggtccgct gtgacacaag aatccctgtt acttctcgac cgtattgatt cggatgattc
4981 ctacgcgagc ctgcggaacg accaggaatt ctgggagccg ctggcccgcc gagccctgga
5041 ggagctcggg ctgccggtgc cgccggtgct gcgggtgccc ggcgagagca ccaaccccgt
5101 actggtcggc gagcccggcc cggtgatcaa gctgttcggc gagcactggt gcggtccgga
5161 gagcctcgcg tcggagtcgg aggcgtacgc ggtcctggcg gacgccccgg tgccggtgcc
5221 ccgcctcctc ggccgcggcg agctgcggcc cggcaccgga gcctggccgt ggccctacct
5281 ggtgatgagc cggatgaccg gcaccacctg gcggtccgcg atggacggca cgaccgaccg
5341 gaacgcgctg ctcgccctgg cccgcgaact cggccgggtg ctcggccggc tgcacagggt
5401 gccgctgacc gggaacaccg tgctcacccc ccattccgag gtcttcccgg aactgctgcg
5461 ggaacgccgc gcggcgaccg tcgaggacca ccgcgggtgg ggctacctct cgccccggct
5521 gctggaccgc ctggaggact ggctgccgga cgtggacacg ctgctggccg gccgcgaacc
5581 ccggttcgtc cacggcgacc tgcacgggac caacatcttc gtggacctgg ccgcgaccga
5641 ggtcaccggg atcgtcgact tcaccgacgt ctatgcggga gactcccgct acagcctggt
5701 gcaactgcat ctcaacgcct tccggggcga ccgcgagatc ctggccgcgc tgctcgacgg
5761 ggcgcagtgg aagcggaccg aggacttcgc ccgcgaactg ctcgccttca ccttcctgca
5821 cgacttcgag gtgttcgagg agaccccgct ggatctctcc ggcttcaccg atccggagga
5881 actggcgcag ttcctctggg ggccgccgga caccgccccc ggcgcctgac gccccgggcc
5941 cgcgccgaag cccgctgctg cgagcccgga gcgggccggc cgacggcggt gcgggcccgg
6001 cggcggacgc tcagcagcgg cgggcgtgaa aggccctggc atcctcgatc atctcctcca
6061 gggtgggtcg gccgggcttc catcccagct cggcaaggat caacgaggac agtcgcacga
6121 cgaagttctt ctggatcgcg cccgtgctgg aagcactcaa cctcgaagcg tgtggttgcg
6181 gagccatcta gcaaccacac gac
//
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