| 质粒名称: | pKO (Kan) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 110086 |
| 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: | pKO (Kan) was a gift from Markus Seeger (Addgene plasmid # 110086 ; http://n2t.net/addgene:110086 ; RRID:Addgene_110086) |
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 (Kan) 5773 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 (Kan)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5773)
TITLE A uniform cloning platform for mycobacterial genetics and protein
production
REFERENCE 2 (bases 1 to 5773)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5773
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 165..195
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind 201..220
/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 1796..1818
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1810..1827
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1811..1827
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(2071..2659)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2151..2170)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 2804..3249
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 3250..4671
/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"
CDS complement(4955..5770)
/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 5678..5697
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 5762..8
/label=pENTR-R
/note="pENTR vectors, reverse primer"
ORIGIN
1 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
61 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttccc cccccccgac
121 gtcaggtggc tagaactaga tatccatgga tccagacccc aggcttgaca ctttatgctt
181 ccggctcgta taatgtgtgg aattgtgagc gctcacaatt cgcatgcggt accaagctga
241 tccgataaca caggaacaga tcttccggag tgtaagaaat gagtctgaaa gaaaaaacac
301 aatctctgtt tgccaacgca tttggctacc ctgccactca caccattcag gcgcctggcc
361 gcgtgaattt gattggtgaa cacaccgact acaacgacgg tttcgttctg ccctgcgcga
421 ttgattatca aaccgtgatc agttgtgcac cacgcgatga ccgtaaagtt cgcgtgatgg
481 cagccgatta tgaaaatcag ctcgacgagt tttccctcga tgcgcccatt gtcgcacatg
541 aaaactatca atgggctaac tacgttcgtg gcgtggtgaa acatctgcaa ctgcgtaaca
601 acagcttcgg cggcgtggac atggtgatca gcggcaatgt gccgcagggt gccgggttaa
661 gttcttccgc ttcactggaa gtcgcggtcg gaaccgtatt gcagcagctt tatcatctgc
721 cgctggacgg cgcacaaatc gcgcttaacg gtcaggaagc agaaaaccag tttgtaggct
781 gtaactgcgg gatcatggat cagctaattt ccgcgctcgg caagaaagat catgccttgc
841 tgatcgattg ccgctcactg gggaccaaag cagtttccat gcccaaaggt gtggctgtcg
901 tcatcatcaa cagtaacttc aaacgtaccc tggttggcag cgaatacaac acccgtcgtg
961 aacagtgcga aaccggtgcg cgtttcttcc agcagccagc cctgcgtgat gtcaccattg
1021 aagagttcaa cgctgttgcg catgaactgg acccgatcgt ggcaaaacgc gtgcgtcata
1081 tactgactga aaacgcccgc accgttgaag ctgccagcgc gctggagcaa ggcgacctga
1141 aacgtatggg cgagttgatg gcggagtctc atgcctctat gcgcgatgat ttcgaaatca
1201 ccgtgccgca aattgacact ctggtagaaa tcgtcaaagc tgtgattggc gacaaaggtg
1261 gcgtacgcat gaccggcggc ggatttggcg gctgtatcgt cgcgctgatc ccggaagaac
1321 tggtgcctgc cgtacagcaa gctgtcgctg aacaatatga agcaaaaaca ggtattaaag
1381 agacttttta cgtttgtaaa ccatcacaag gagcaggaca gtgctgaacg aaactcctgc
1441 agcgcgtagt acctcacacc gtcctgcggg ttgacgtatg aggcttggat cctcccgctg
1501 cgctgagtct tcagcgatcc ccatccgcga cgtgccaact aggtctcctc tcgtcgtgaa
1561 caaggctacc gggttgcaac tcctgtgcaa ctctcaggct tcaacgcgct tctacgacct
1621 gcaatttctt tccacttaga ggatgcagcc gagagggggt aaaaacctcc cattcgccat
1681 tcaggctgcg caactgttgg gaagggcgtt tcggtgcggg cctcttcgct attacgccag
1741 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
1801 tcacgacgtt gtaaaacgac ggccagtgag cgcgacgtaa taagttgaag agcatctgct
1861 cttcagcatc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaacatg
1921 gtcatagctg tttccttgcg tattgggcgc tctccgcttc ctcgctcact gactcgctgc
1981 gctcggtcgt tcgggtaaag cctggggtgc ctaatgagca aaaggccagc aaaaggccag
2041 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
2101 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
2161 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
2221 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
2281 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
2341 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
2401 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
2461 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
2521 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
2581 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
2641 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
2701 gaacgaaggc gcgcctgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
2761 cgagtcagtg agcgaggaag cggaagatcg ggcccagccg gcccacatat acctgccgtt
2821 cactattatt tagtgaaatg agatattatg atattttctg aattgtgatt aaaaaggcaa
2881 ctttatgccc atgcaacaga aactataaaa aatacagaga atgaaaagaa acagatagat
2941 tttttagttc tttaggcccg tagtctgcaa atccttttat gattttctat caaacaaaag
3001 aggaaaatag accagttgca atccaaacga gagtctaata gaatgaggtc gaaaagtaaa
3061 tcgcgcgggt ttgttactga taaagcaggc aagacctaaa atgtgtaaag ggcaaagtgt
3121 atactttggc gtcacccctt acatatttta ggtctttttt tattgtgcgt aactaacttg
3181 ccatcttcaa acaggagggc tggaagaagc agaccgctaa cacagtacat aaaaaaggag
3241 acatgaacga tgaacatcaa aaagtttgca aaacaagcaa cagtattaac ctttactacc
3301 gcactgctgg caggaggcgc aactcaagcg tttgcgaaag aaacgaacca aaagccatat
3361 aaggaaacat acggcatttc ccatattaca cgccatgata tgctgcaaat ccctgaacag
3421 caaaaaaatg aaaaatataa agttcctgag ttcgattcgt ccacaattaa aaatatctct
3481 tctgcaaaag gcctggacgt ttgggacagc tggccattac aaaacactga cggcactgtc
3541 gcaaactatc acggctacca catcgtcttt gcattagccg gagatcctaa aaatgcggat
3601 gacacatcga tttacatgtt ctatcaaaaa gtcggcgaaa cttctattga cagctggaaa
3661 aacgctggcc gcgtctttaa agacagcgac aaattcgatg caaatgattc tatcctaaaa
3721 gaccaaacac aagaatggtc aggttcagcc acatttacat ctgacggaaa aatccgttta
3781 ttctacactg atttctccgg taaacattac ggcaaacaaa cactgacaac tgcacaagtt
3841 aacgtatcag catcagacag ctctttgaac atcaacggtg tagaggatta taaatcaatc
3901 tttgacggtg acggaaaaac gtatcaaaat gtacagcagt tcatcgatga aggcaactac
3961 agctcaggcg acaaccatac gctgagagat cctcactacg tagaagataa aggccacaaa
4021 tacttagtat ttgaagcaaa cactggaact gaagatggct accaaggcga agaatcttta
4081 tttaacaaag catactatgg caaaagcaca tcattcttcc gtcaagaaag tcaaaaactt
4141 ctgcaaagcg ataaaaaacg cacggctgag ttagcaaacg gcgctctcgg tatgattgag
4201 ctaaacgatg attacacact gaaaaaagtg atgaaaccgc tgattgcatc taacacagta
4261 acagatgaaa ttgaacgcgc gaacgtcttt aaaatgaacg gcaaatggta cctgttcact
4321 gactcccgcg gatcaaaaat gacgattgac ggcattacgt ctaacgatat ttacatgctt
4381 ggttatgttt ctaattcttt aactggccca tacaagccgc tgaacaaaac tggccttgtg
4441 ttaaaaatgg atcttgatcc taacgatgta acctttactt actcacactt cgctgtacct
4501 caagcgaaag gaaacaatgt cgtgattaca agctatatga caaacagagg attctacgca
4561 gacaaacaat caacgtttgc gcctagcttc ctgctgaaca tcaaaggcaa gaaaacatct
4621 gttgtcaaag acagcatcct tgaacaagga caattaacag ttaacaaata aaaacgcaaa
4681 agaaaatgcc gatatcctat tggcattgcg gccgcggtct cggcggtggg tcgtctagag
4741 aaagtgaggg agccacggtt ggtgagagct ttgttgtagg tggaccagtt ggtgattttg
4801 aacttttgct ttgccacgga atggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc
4861 aactcagcaa aagttcgatt tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc
4921 tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat
4981 gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa agccgtttct
5041 gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt
5101 ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa
5161 ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagat
5221 tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac
5281 tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat
5341 cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca
5401 gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt
5461 tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga
5521 tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat
5581 cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatgg ggcttcccat
5641 acaagcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat
5701 ataaatcagc atccatgttg gaatttaatc gcggcctcga gcaagacgtt tcccgttgaa
5761 tatggctcat aac
//
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