| 质粒名称: | pKO (Apr) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 110088 |
| 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: | 安普霉素 |
| 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 (Apr) was a gift from Markus Seeger (Addgene plasmid # 110088 ; http://n2t.net/addgene:110088 ; RRID:Addgene_110088) |
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 (Apr) 5685 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 (Apr)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5685)
TITLE A uniform cloning platform for mycobacterial genetics and protein
production
REFERENCE 2 (bases 1 to 5685)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5685
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 105..135
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
protein_bind 141..160
/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 1736..1758
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 1750..1767
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 1751..1767
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(2011..2599)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2091..2110)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
promoter 2744..3189
/gene="sacR"
/label=sacB promoter
/note="sacB promoter and control region"
CDS 3190..4611
/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 4796..5596
/codon_start=1
/gene="aac(3)IV"
/product="aminoglycoside 3-N-acetyltransferase type IV
"
/label=ApmR
/note="confers resistance to apramycin (Paget and Davies,
1996)"
/translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
ORIGIN
1 tttcttcagc tcgctgatga tatgctgacg ctcaatgccg tttggcctcc gactaacgaa
61 aatcccgcat ttggacggct gatccatgga tccagacccc aggcttgaca ctttatgctt
121 ccggctcgta taatgtgtgg aattgtgagc gctcacaatt cgcatgcggt accaagctga
181 tccgataaca caggaacaga tcttccggag tgtaagaaat gagtctgaaa gaaaaaacac
241 aatctctgtt tgccaacgca tttggctacc ctgccactca caccattcag gcgcctggcc
301 gcgtgaattt gattggtgaa cacaccgact acaacgacgg tttcgttctg ccctgcgcga
361 ttgattatca aaccgtgatc agttgtgcac cacgcgatga ccgtaaagtt cgcgtgatgg
421 cagccgatta tgaaaatcag ctcgacgagt tttccctcga tgcgcccatt gtcgcacatg
481 aaaactatca atgggctaac tacgttcgtg gcgtggtgaa acatctgcaa ctgcgtaaca
541 acagcttcgg cggcgtggac atggtgatca gcggcaatgt gccgcagggt gccgggttaa
601 gttcttccgc ttcactggaa gtcgcggtcg gaaccgtatt gcagcagctt tatcatctgc
661 cgctggacgg cgcacaaatc gcgcttaacg gtcaggaagc agaaaaccag tttgtaggct
721 gtaactgcgg gatcatggat cagctaattt ccgcgctcgg caagaaagat catgccttgc
781 tgatcgattg ccgctcactg gggaccaaag cagtttccat gcccaaaggt gtggctgtcg
841 tcatcatcaa cagtaacttc aaacgtaccc tggttggcag cgaatacaac acccgtcgtg
901 aacagtgcga aaccggtgcg cgtttcttcc agcagccagc cctgcgtgat gtcaccattg
961 aagagttcaa cgctgttgcg catgaactgg acccgatcgt ggcaaaacgc gtgcgtcata
1021 tactgactga aaacgcccgc accgttgaag ctgccagcgc gctggagcaa ggcgacctga
1081 aacgtatggg cgagttgatg gcggagtctc atgcctctat gcgcgatgat ttcgaaatca
1141 ccgtgccgca aattgacact ctggtagaaa tcgtcaaagc tgtgattggc gacaaaggtg
1201 gcgtacgcat gaccggcggc ggatttggcg gctgtatcgt cgcgctgatc ccggaagaac
1261 tggtgcctgc cgtacagcaa gctgtcgctg aacaatatga agcaaaaaca ggtattaaag
1321 agacttttta cgtttgtaaa ccatcacaag gagcaggaca gtgctgaacg aaactcctgc
1381 agcgcgtagt acctcacacc gtcctgcggg ttgacgtatg aggcttggat cctcccgctg
1441 cgctgagtct tcagcgatcc ccatccgcga cgtgccaact aggtctcctc tcgtcgtgaa
1501 caaggctacc gggttgcaac tcctgtgcaa ctctcaggct tcaacgcgct tctacgacct
1561 gcaatttctt tccacttaga ggatgcagcc gagagggggt aaaaacctcc cattcgccat
1621 tcaggctgcg caactgttgg gaagggcgtt tcggtgcggg cctcttcgct attacgccag
1681 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
1741 tcacgacgtt gtaaaacgac ggccagtgag cgcgacgtaa taagttgaag agcatctgct
1801 cttcagcatc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaacatg
1861 gtcatagctg tttccttgcg tattgggcgc tctccgcttc ctcgctcact gactcgctgc
1921 gctcggtcgt tcgggtaaag cctggggtgc ctaatgagca aaaggccagc aaaaggccag
1981 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
2041 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
2101 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
2161 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
2221 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
2281 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
2341 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
2401 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
2461 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
2521 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
2581 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
2641 gaacgaaggc gcgcctgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
2701 cgagtcagtg agcgaggaag cggaagatcg ggcccagccg gcccacatat acctgccgtt
2761 cactattatt tagtgaaatg agatattatg atattttctg aattgtgatt aaaaaggcaa
2821 ctttatgccc atgcaacaga aactataaaa aatacagaga atgaaaagaa acagatagat
2881 tttttagttc tttaggcccg tagtctgcaa atccttttat gattttctat caaacaaaag
2941 aggaaaatag accagttgca atccaaacga gagtctaata gaatgaggtc gaaaagtaaa
3001 tcgcgcgggt ttgttactga taaagcaggc aagacctaaa atgtgtaaag ggcaaagtgt
3061 atactttggc gtcacccctt acatatttta ggtctttttt tattgtgcgt aactaacttg
3121 ccatcttcaa acaggagggc tggaagaagc agaccgctaa cacagtacat aaaaaaggag
3181 acatgaacga tgaacatcaa aaagtttgca aaacaagcaa cagtattaac ctttactacc
3241 gcactgctgg caggaggcgc aactcaagcg tttgcgaaag aaacgaacca aaagccatat
3301 aaggaaacat acggcatttc ccatattaca cgccatgata tgctgcaaat ccctgaacag
3361 caaaaaaatg aaaaatataa agttcctgag ttcgattcgt ccacaattaa aaatatctct
3421 tctgcaaaag gcctggacgt ttgggacagc tggccattac aaaacactga cggcactgtc
3481 gcaaactatc acggctacca catcgtcttt gcattagccg gagatcctaa aaatgcggat
3541 gacacatcga tttacatgtt ctatcaaaaa gtcggcgaaa cttctattga cagctggaaa
3601 aacgctggcc gcgtctttaa agacagcgac aaattcgatg caaatgattc tatcctaaaa
3661 gaccaaacac aagaatggtc aggttcagcc acatttacat ctgacggaaa aatccgttta
3721 ttctacactg atttctccgg taaacattac ggcaaacaaa cactgacaac tgcacaagtt
3781 aacgtatcag catcagacag ctctttgaac atcaacggtg tagaggatta taaatcaatc
3841 tttgacggtg acggaaaaac gtatcaaaat gtacagcagt tcatcgatga aggcaactac
3901 agctcaggcg acaaccatac gctgagagat cctcactacg tagaagataa aggccacaaa
3961 tacttagtat ttgaagcaaa cactggaact gaagatggct accaaggcga agaatcttta
4021 tttaacaaag catactatgg caaaagcaca tcattcttcc gtcaagaaag tcaaaaactt
4081 ctgcaaagcg ataaaaaacg cacggctgag ttagcaaacg gcgctctcgg tatgattgag
4141 ctaaacgatg attacacact gaaaaaagtg atgaaaccgc tgattgcatc taacacagta
4201 acagatgaaa ttgaacgcgc gaacgtcttt aaaatgaacg gcaaatggta cctgttcact
4261 gactcccgcg gatcaaaaat gacgattgac ggcattacgt ctaacgatat ttacatgctt
4321 ggttatgttt ctaattcttt aactggccca tacaagccgc tgaacaaaac tggccttgtg
4381 ttaaaaatgg atcttgatcc taacgatgta acctttactt actcacactt cgctgtacct
4441 caagcgaaag gaaacaatgt cgtgattaca agctatatga caaacagagg attctacgca
4501 gacaaacaat caacgtttgc gcctagcttc ctgctgaaca tcaaaggcaa gaaaacatct
4561 gttgtcaaag acagcatcct tgaacaagga caattaacag ttaacaaata aaaacgcaaa
4621 agaaaatgcc gatatcctat tggcattgcg gccgcggtct cggcggtggg tcgtctagaa
4681 ggatcttcac ctagatcctt ttggttcatg tgcagctcca tcagcaaaag gggatgataa
4741 gtttatcacc accgactatt tgcaacagtg ccgttgatcg tgctatgatc gactgatgtc
4801 atcagcggtg gagtgcaatg tcgtgcaata cgaatggcga aaagccgagc tcatcggtca
4861 gcttctcaac cttggggtta cccccggcgg tgtgctgctg gtccacagct ccttccgtag
4921 cgtccggccc ctcgaagatg ggccacttgg actgatcgag gccctgcgtg ctgcgctggg
4981 tccgggaggg acgctcgtca tgccctcgtg gtcaggtctg gacgacgagc cgttcgatcc
5041 tgccacgtcg cccgttacac cggaccttgg agttgtctct gacacattct ggcgcctgcc
5101 aaatgtaaag cgcagcgccc atccatttgc ctttgcggca gcggggccac aggcagagca
5161 gatcatctct gatccattgc ccctgccacc tcactcgcct gcaagcccgg tcgcccgtgt
5221 ccatgaactc gatgggcagg tacttctcct cggcgtggga cacgatgcca acacgacgct
5281 gcatcttgcc gagttgatgg caaaggttcc ctatggggtg ccgagacact gcaccattct
5341 tcaggatggc aagttggtac gcgtcgatta tctcgagaat gaccactgct gtgagcgctt
5401 tgccttggcg gacaggtggc tcaaggaaaa gagccttcag aaggaaggtc cagtcggtca
5461 tgcctttgct cggttgatcc gctcccgcga cattgtggcg acagccctgg gtcaactggg
5521 ccgagatccg ttgatcttcc tgcatccgcc agaggcggga tgcgaagaat gcgatgccgc
5581 tcgccagtcg attggctgag ctcatgagcg gagaacgaga tgacgttgga ggggcaaggt
5641 cgcgctgatt gctggggcaa cacgtggagc ggatcgggga ttgtc
//
