| 质粒名称: | pKOdeltagalK (Hyg) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 110090 |
| 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: | pKOdeltagalK (Hyg) was a gift from Markus Seeger (Addgene plasmid # 110090 ; http://n2t.net/addgene:110090 ; RRID:Addgene_110090) |
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 (Hyg) 4864 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Analogous to pKO but without the negative selection gene galK..
ACCESSION .
VERSION .
KEYWORDS pKOdeltagalK (Hyg)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4864)
TITLE A uniform cloning platform for mycobacterial genetics and protein
production
REFERENCE 2 (bases 1 to 4864)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4864
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 1785..1807
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1799..1816
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1800..1816
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(2060..2648)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2140..2159)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 2793..3238
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 3239..4660
/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 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
61 ttcatccata gttgcctgac tccccgacgt ggccgaccag cccgtcatcg tcaacgcctg
121 atccgcggtg cggacaggcc gtgtcgcgac cggccgtgcg gaattaagcc ggcccgtacc
181 ctgtgaatag aggtccgctg tgacacaaga atccctgtta cttctcgacc gtattgattc
241 ggatgattcc tacgcgagcc tgcggaacga ccaggaattc tgggagccgc tggcccgccg
301 agccctggag gagctcgggc tgccggtgcc gccggtgctg cgggtgcccg gcgagagcac
361 caaccccgta ctggtcggcg agcccggccc ggtgatcaag ctgttcggcg agcactggtg
421 cggtccggag agcctcgcgt cggagtcgga ggcgtacgcg gtcctggcgg acgccccggt
481 gccggtgccc cgcctcctcg gccgcggcga gctgcggccc ggcaccggag cctggccgtg
541 gccctacctg gtgatgagcc ggatgaccgg caccacctgg cggtccgcga tggacggcac
601 gaccgaccgg aacgcgctgc tcgccctggc ccgcgaactc ggccgggtgc tcggccggct
661 gcacagggtg ccgctgaccg ggaacaccgt gctcaccccc cattccgagg tcttcccgga
721 actgctgcgg gaacgccgcg cggcgaccgt cgaggaccac cgcgggtggg gctacctctc
781 gccccggctg ctggaccgcc tggaggactg gctgccggac gtggacacgc tgctggccgg
841 ccgcgaaccc cggttcgtcc acggcgacct gcacgggacc aacatcttcg tggacctggc
901 cgcgaccgag gtcaccggga tcgtcgactt caccgacgtc tatgcgggag actcccgcta
961 cagcctggtg caactgcatc tcaacgcctt ccggggcgac cgcgagatcc tggccgcgct
1021 gctcgacggg gcgcagtgga agcggaccga ggacttcgcc cgcgaactgc tcgccttcac
1081 cttcctgcac gacttcgagg tgttcgagga gaccccgctg gatctctccg gcttcaccga
1141 tccggaggaa ctggcgcagt tcctctgggg gccgccggac accgcccccg gcgcctgacg
1201 ccccgggccc gcgccgaagc ccgctgctgc gagcccggag cgggccggcc gacggcggtg
1261 cgggcccggc ggcggacgct cagcagcggc gggcgtgaaa ggccctggca tcctcgatca
1321 tctcctccag ggtgggtcgg ccgggcttcc atcccagctc ggcaaggatc aacgaggaca
1381 gtcgcacgac gaagttcttc tggatcgcgc ccgtgctgga agcactcaac ctcgaagcgt
1441 gtggttgcgg agccatctag caaccacacg accatggatc ctcccgctgc gctgagtctt
1501 cagcgatccc catccgcgac gtgccaacta ggtctcctct cgtcgtgaac aaggctaccg
1561 ggttgcaact cctgtgcaac tctcaggctt caacgcgctt ctacgacctg caatttcttt
1621 ccacttagag gatgcagccg agagggggta aaaacctccc attcgccatt caggctgcgc
1681 aactgttggg aagggcgttt cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
1741 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
1801 taaaacgacg gccagtgagc gcgacgtaat aagttgaaga gcatctgctc ttcagcatca
1861 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaacatgg tcatagctgt
1921 ttccttgcgt attgggcgct ctccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
1981 cgggtaaagc ctggggtgcc taatgagcaa aaggccagca aaaggccagg aaccgtaaaa
2041 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
2101 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
2161 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
2221 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
2281 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
2341 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
2401 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
2461 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
2521 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
2581 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
2641 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaggcg
2701 cgcctgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
2761 gcgaggaagc ggaagatcgg gcccagccgg cccacatata cctgccgttc actattattt
2821 agtgaaatga gatattatga tattttctga attgtgatta aaaaggcaac tttatgccca
2881 tgcaacagaa actataaaaa atacagagaa tgaaaagaaa cagatagatt ttttagttct
2941 ttaggcccgt agtctgcaaa tccttttatg attttctatc aaacaaaaga ggaaaataga
3001 ccagttgcaa tccaaacgag agtctaatag aatgaggtcg aaaagtaaat cgcgcgggtt
3061 tgttactgat aaagcaggca agacctaaaa tgtgtaaagg gcaaagtgta tactttggcg
3121 tcacccctta catattttag gtcttttttt attgtgcgta actaacttgc catcttcaaa
3181 caggagggct ggaagaagca gaccgctaac acagtacata aaaaaggaga catgaacgat
3241 gaacatcaaa aagtttgcaa aacaagcaac agtattaacc tttactaccg cactgctggc
3301 aggaggcgca actcaagcgt ttgcgaaaga aacgaaccaa aagccatata aggaaacata
3361 cggcatttcc catattacac gccatgatat gctgcaaatc cctgaacagc aaaaaaatga
3421 aaaatataaa gttcctgagt tcgattcgtc cacaattaaa aatatctctt ctgcaaaagg
3481 cctggacgtt tgggacagct ggccattaca aaacactgac ggcactgtcg caaactatca
3541 cggctaccac atcgtctttg cattagccgg agatcctaaa aatgcggatg acacatcgat
3601 ttacatgttc tatcaaaaag tcggcgaaac ttctattgac agctggaaaa acgctggccg
3661 cgtctttaaa gacagcgaca aattcgatgc aaatgattct atcctaaaag accaaacaca
3721 agaatggtca ggttcagcca catttacatc tgacggaaaa atccgtttat tctacactga
3781 tttctccggt aaacattacg gcaaacaaac actgacaact gcacaagtta acgtatcagc
3841 atcagacagc tctttgaaca tcaacggtgt agaggattat aaatcaatct ttgacggtga
3901 cggaaaaacg tatcaaaatg tacagcagtt catcgatgaa ggcaactaca gctcaggcga
3961 caaccatacg ctgagagatc ctcactacgt agaagataaa ggccacaaat acttagtatt
4021 tgaagcaaac actggaactg aagatggcta ccaaggcgaa gaatctttat ttaacaaagc
4081 atactatggc aaaagcacat cattcttccg tcaagaaagt caaaaacttc tgcaaagcga
4141 taaaaaacgc acggctgagt tagcaaacgg cgctctcggt atgattgagc taaacgatga
4201 ttacacactg aaaaaagtga tgaaaccgct gattgcatct aacacagtaa cagatgaaat
4261 tgaacgcgcg aacgtcttta aaatgaacgg caaatggtac ctgttcactg actcccgcgg
4321 atcaaaaatg acgattgacg gcattacgtc taacgatatt tacatgcttg gttatgtttc
4381 taattcttta actggcccat acaagccgct gaacaaaact ggccttgtgt taaaaatgga
4441 tcttgatcct aacgatgtaa cctttactta ctcacacttc gctgtacctc aagcgaaagg
4501 aaacaatgtc gtgattacaa gctatatgac aaacagagga ttctacgcag acaaacaatc
4561 aacgtttgcg cctagcttcc tgctgaacat caaaggcaag aaaacatctg ttgtcaaaga
4621 cagcatcctt gaacaaggac aattaacagt taacaaataa aaacgcaaaa gaaaatgccg
4681 atatcctatt ggcattgcgg ccgcggtctc ggcggtgggt cgtctagagg tctgacgctc
4741 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
4801 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
4861 cttg
//
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