| 质粒名称: | pRPF215 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 106377 |
| 存储实验室: | Robert Fagan, Neil Fairweather |
|
载体骨架基本信息 |
|
| 载体名称: | pMTL960 |
| 载体出品公司: | Clontech |
| 空载体大小: | 6191bp |
| 完整质粒大小: | 9231bp |
| 载体类型: | E. coli - C. difficile shuttle vector |
| 载体抗性: | Chloramphenicol, 15 μg/mL |
| 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: | Ptet-himar1-Ter(slpA)-transposon |
| 插入大小: | 3040bp |
| 基因突变: | Codon optimised for C. difficile |
| 启动子: | Ptet |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | KpnI (not destroyed) |
| 3’克隆位点: | BstXI (not destroyed) |
| 5’测序引物 | CACCTCCTTTTTGACTTTAAGCCTACGAATACC |
| 3’测序引物 |
CACCGACGAGCAAGGCAAGACCG
(Common Sequencing Primers)
|
|
文献引用方法 |
|
| 材料和方法部分: | pRPF215 was a gift from Robert Fagan & Neil Fairweather (Addgene plasmid # 106377 ; http://n2t.net/addgene:106377 ; RRID:Addgene_106377) |
| 参考文献部分: | High-throughput analysis of gene essentiality and sporulation in Clostridium difficile. Dembek M, Barquist L, Boinett CJ, Cain AK, Mayho M, Lawley TD, Fairweather NF, Fagan RP. MBio. 2015 Feb 24;6(2):e02383. doi: 10.1128/mBio.02383-14. 10.1128/mBio.02383-14 PubMed 25714712 |
买家导航
pRPF215载体质粒基本信息
pRPF215质粒图谱载体图谱和pRPF215载体序列质粒序列多克隆位点信息
pRPF215质粒载体简介
pRPF215质粒序列载体序列
LOCUS pRPF215 9231 bp DNA circular SYN 14-APR-2025
DEFINITION C. difficile transposon mutagenesis system. Shuttle vector
containing tet-inducible Himar1 transposase and custom ErmR
transposon.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9231)
AUTHORS Dembek M, Barquist L, Boinett CJ, Cain AK, Mayho M, Lawley TD,
Fairweather NF, Fagan RP
TITLE High-throughput analysis of gene essentiality and sporulation in
Clostridium difficile.
JOURNAL MBio. 2015 Feb 24;6(2):e02383. doi: 10.1128/mBio.02383-14.
PUBMED 25714712
REFERENCE 2 (bases 1 to 9231)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "MBio.";
date: "2015-02-24"; volume: "6(2):e02383. doi"; pages: "
10.1128/mBio.02383-14"
COMMENT Sequence Label: pRPF215
FEATURES Location/Qualifiers
source 1..9231
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(2710..3336)
/codon_start=1
/gene="tetR"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind 3241..3260
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
protein_bind 3443..3461
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
CDS complement(5942..6313)
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT 6346..6455
/note="incP origin of transfer"
primer_bind complement(7646..7663)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7817..8405)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7897..7916)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cctctatcat ttattttagt ttcccataca tctaaaattt tagtatctaa taaagattct
61 atcatatttg aaattctttg ctgtgttact cctaaatctt ttgctaattc ctcttgatta
121 attcttaata tatttccttt tattttatca tctttgacca ataaatcttg cttatatctc
181 atatgagtat ataatcttaa ttcttcagtt gaaattattc cccaaattac tgccatggta
241 acaaagtaac taatagtata ttgtttatct atattacatc taacttcttt tacattgtat
301 gtatctttta ttcctagctt tgtaacacct tttgtttttg taatataatc attatgttga
361 agttctttta atgtttctct taatgtcttt tcactcataa caatgttttt aactttcttc
421 ttacgtttat aggttatcag ctccattata tcgtctatgt ttaattctct atctttattg
481 ttaagtaaca cattatagat aaacaattga ttaccagtca ttattttaac ccctttccta
541 ttcttatatt tcaaatcctt tgagtgctta tgtggcatat tgaacaaagt atcttcatca
601 ctatatataa aatctgattc tattttattc caacaatcac tattaaactc acaacctgta
661 caaccaaaat taaccgtttt aagcttctca tacatatact tgacttgata ttctagctca
721 ctatttgaaa gactattttg tgatatatag ttccattctc tagcaacgga tattacttga
781 cttaaacttt tattcctcaa tcttaaagtc acaactattt tttgcaaatc taggttttta
841 tgtccatctt tactaccatt ttttaaaata tctactatac aatttggtag cttactatct
901 aatatatatt ttgtattagt attacctact gtttttacat ctttgcaata ttttttagct
961 aacttttcta tgtcatatct atatatattt ttgtcttggt gtattatttt tacttgttta
1021 gtagtatttt taatattata tgtataaggt actcttaata cttgagtaga taaatttgct
1081 tgtttatctg cacccaattt atttattaat gtatttgtaa cttctgatac taacttaatg
1141 ttattagttt tattaattag cacataaaca tggaatccat ttccactatc aactaggata
1201 tgataatgta atcctatctt agtaaataaa ttaattatat ctttataatt aaaaccttgt
1261 cctaattctt ttttgtcaaa atcaaatcct aatgcatatc tatttattaa gttttcattg
1321 gttcctgctt ttccatcagt tgtacatagg ttaaaaaatg tattttcata ataaatattt
1381 tttgcgaagg taaattctac tatatcatct atatttttaa aaaaagttgt tttactatat
1441 tctttaaatt cttctccgag ttctaattgt ttactattac ttttattatt ttgaaatact
1501 ctaacatatt cattatcaat aagattattt ccttgtccta taccatctct aaatactgcc
1561 attagaaatt ttttcaactt atattttgaa tctaattgtt ccatatattt attatctcct
1621 tatctagttt ttttaagaga taacactaca ttaaatatat acttattgac taatcaactt
1681 ttacaactat tgaataattt tagcttagta tgtataataa tattgttaag atgctaaaat
1741 tgatagcaat attattatat aactacagat acttaatcag ttagaccgat attggagttg
1801 gtagctccat aaactcaatt ggtagttgag ttgtcattgt ctagtagtat tatctcattt
1861 tgttgttatc ttaaaatatt attttaaaaa tactctgctt ggtcggctta gggtattttt
1921 tttgtttaat tccaatattg gttcaaactc ctgtgtaata cttttgacac ttccatctgt
1981 attgaattga cagttgatat ggtatctcat tattggcttg ttcttatgtt tttctatgta
2041 tttatttttt actttcaaat aatttgtatt tccattttta gttatgttta acacactatc
2101 cttattactc aataaaatta ttttttgttt taacatatga tttatagaat ctaaaaattg
2161 atattgactt ataatagtac tttctacctc tctagctttt tctagtgttt caaattcttc
2221 aaactctgct atattgctat agtatagctt atcattaata atattatctg acaactttaa
2281 ttccatttta taaaacaatt ccaagttatg tataaaattt ttaatgttac aacatataga
2341 cttattattt gttcttatat ggtatctcca tttactactt ctagctctca tgttatatgc
2401 acaactactc ccaacataaa catatattgc ttgttctaac tcatattttt gtaaaatttc
2461 aacatcatat ccagtcaaat cgtactctct cttatataca aatggttttt ctaatttttt
2521 gttcaataaa tctcctccta tttcttactt aatttagcta aaaatctatt tatatttcac
2581 ctcctttttg actttaagcc tacgaatacc atatttatga ttataacatc ttctattttt
2641 cccaaatcct tacatctccc ctaattttta gacttaaggg cgaattctgc atcaagctag
2701 cttggtacct taagacccac tttcacattt aagttgtttt tctaatccgc atatgatcaa
2761 ttcaaggccg aataagaagg ctggctctgc accttggtga tcaaataatt cgatagcttg
2821 tcgtaataat ggcggcatac tatcagtagt aggtgtttcc ctttcttctt tagcgacttg
2881 atgctcttga tcttccaata cgcaacctaa agtaaaatgc cccacagcgc tgagtgcata
2941 taatgcattc tctagtgaaa aaccttgttg gcataaaaag gctaattgat tttcgagagt
3001 ttcatactgt ttttctgtag gccgtgtacc taaatgtact tttgctccat cgcgatgact
3061 tagtaaagca catctaaaac ttttagcgtt attacgtaaa aaatcttgcc agctttcccc
3121 ttctaaaggg caaaagtgag tatggtgcct atctaacatc tcaatggcta aggcgtcgag
3181 caaagcccgc ttatttttta catgccaata caatgtaggc tgctctacac ctagcttctg
3241 ggcgagttta cgggttgtta aaccttcgat tccgacctca ttaagcagct ctaatgcgct
3301 gttaatcact ttacttttat ctaatctaga catcattaat tcctcctttt tgttgacatt
3361 atatcattga tagagttatt tgtcaaacta gttttttatt tcgatgccct ggacttcatg
3421 aaaaactaaa aaaaatattg acactctatc attgatagag tataattaaa ataagcttga
3481 tcgtagcgtt aacagatctg agctcaataa tactaggagg tttttatgga aaaaaaagaa
3541 tttagagtat taataaaata ttgtttttta aaaggaaaga atacagtaga agcaaaaaca
3601 tggttagata atgaatttcc agatagtgca ccaggaaaaa gtacaataat agattggtat
3661 gcaaaattta aaagaggtga aatgagtact gaagatggtg aaagaagtgg tagaccaaaa
3721 gaagtagtaa ctgatgaaaa tataaaaaaa atacataaaa tgatacttaa tgatagaaaa
3781 atgaaattaa tagaaatagc agaagcatta aaaataagta aagaaagagt aggacatata
3841 atacatcaat atttagatat gagaaaatta tgtgcaaaat gggttccaag agaattaaca
3901 tttgatcaaa aacaacaaag agtagatgat tctgaaagat gtttacaatt attaacaaga
3961 aatacaccag aattttttag aagatatgta acaatggatg aaacatggtt acatcattat
4021 acaccagaaa gtaatagaca aagtgcagaa tggacagcaa caggtgaacc atctccaaaa
4081 agaggaaaaa cacaaaaatc tgctggaaaa gtaatggcaa gtgtattttg ggatgcacat
4141 ggaataatat ttatagatta tttagaaaaa ggtaaaacaa taaatagtga ttattatatg
4201 gcattattag aaagattaaa agtagaaata gctgcaaaaa gaccacatat gaaaaaaaaa
4261 aaagtattat ttcatcaaga taatgcacca tgtcataaaa gtttaagaac aatggcaaaa
4321 atacatgaat taggatttga attattacca catccaccat atagtccaga tttagcacca
4381 agtgattttt ttttatttag tgatttaaaa agaatgttag caggaaaaaa atttggttgt
4441 aatgaagaag taatagcaga aacagaagca tattttgaag caaaaccaaa agaatattat
4501 caaaatggaa taaaaaaatt agaaggtaga tataatagat gtatagcatt agaaggtaat
4561 tatgtagaat aataaaggcc tcggatccta taagttttaa taaaacttta aatagaaaaa
4621 ggcttctctc atgagaagtc ttttttattt aaaataaata taaaataaaa tagaggctat
4681 aaatagcctc tattttatgt gagaaatccc taaataaaaa gatgccagtg tgctggtaac
4741 aggttggctg ataagtcccc ggtctgacag aagcaaactt aagagtgtgt tgatagtgca
4801 gtatcttaaa attttgtata ataggaattg aagttaaatt agatgctaaa aatttgtaat
4861 taagaaggag tgattacatg aacaaaaata taaaatattc tcaaaacttt ttaacgagtg
4921 aaaaagtact caaccaaata ataaaacaat tgaatttaaa agaaaccgat accgtttacg
4981 aaattggaac aggtaaaggg catttaacga cgaaactggc taaaataagt aaacaggtaa
5041 cgtctattga attagacagt catctattca acttatcgtc agaaaaatta aaactgaata
5101 ctcgtgtcac tttaattcac caagatattc tacagtttca attccctaac aaacagaggt
5161 ataaaattgt tgggagtatt ccttaccatt taagcacaca aattattaaa aaagtggttt
5221 ttgaaagcca tgcgtctgac atctatctga ttgttgaaga aggattctac aagcgtacct
5281 tggatattca ccgaacacta gggttgctct tgcacactca agtctcgatt cagcaattgc
5341 ttaagctgcc agcggaatgc tttcatccta aaccaaaagt aaacagtgtc ttaataaaac
5401 ttacccgcca taccacagat gttccagata aatattggaa gctatatacg tactttgttt
5461 caaaatgggt caatcgagaa tatcgtcaac tgtttactaa aaatcagttt catcaagcaa
5521 tgaaacacgc caaagtaaac aatttaagta ccgttactta tgagcaagta ttgtctattt
5581 ttaatagtta tctattattt aacgggagga aataattcta tgagtcgctt ttgtaaattt
5641 ggaaagttac acgttactaa aggcataaaa ataagaagcc tgcaaatgca ggcttcttat
5701 ttttatgtgt tagaccgggg acttatcagc caacctgtta ccagtgtgct ggaattcgcc
5761 cttaggctgc gctagggacc tctttagctc cttggaagct gtcagtagta ggggatcggt
5821 cttgccttgc tcgtcggtga tgtacttcac cagctccgcg aagtcgctct tcttgatgga
5881 gcgcatgggg acgtgcttgg caatcacgcg caccccccgg ccgttttagc ggctaaaaaa
5941 gtcatggctc tgccctcggg cggaccacgc ccatcatgac cttgccaagc tcgtcctgct
6001 tctcttcgat cttcgccagc agggcgagga tcgtggcatc accgaaccgc gccgtgcgcg
6061 ggtcgtcggt gagccagagt ttcagcaggc cgcccaggcg gcccaggtcg ccattgatgc
6121 gggccagctc gcggacgtgc tcatagtcca cgacgcccgt gattttgtag ccctggccga
6181 cggccagcag gtaggccgac aggctcatgc cggccgccgc cgccttttcc tcaatcgctc
6241 ttcgttcgtc tggaaggcag tacaccttga taggtgggct gcccttcctg gttggcttgg
6301 tttcatcagc catccgcttg ccctcatctg ttacgccggc ggtagccggc cagcctcgca
6361 gagcaggatt cccgttgagc accgccaggt gcgaataagg gacagtgaag aaggaacacc
6421 cgctcgcggg tgggcctact tcacctatcc tgcccggctg acgccgttgg atacaccaag
6481 gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg tgcgaaaaag gatggatata
6541 ccgaaaaaat cgctataatg accccgaagc aggaagggcg aattctgcag atctgaccgg
6601 tctctgaaaa tataaaaacc acagattgat actaaaacct tggttgtgtt gcttttcggg
6661 gcttaaatca aggaaaaatc cttgttttaa gcctttcaaa aagaaacaca aggtctttgt
6721 actaacctgt ggttatgtat aaaattgtag attttagggt aacaaaaaac accgtatttc
6781 tacgatgttt ttgcttaaat acttgttttt agttacagac aaacctgaag ttaactattt
6841 atcaattcct gcaattcgtt tacaaaacgg caaatgtgaa atccgtcaca tactgcgtga
6901 tgaacttgaa ttgccaaagg aagtataatt ttgttatctt ctttataata tttccccata
6961 gtaaaaatag gaatcaaata atcatatcct ttctgcaaat tcagattaaa gccatcgaag
7021 gttgaccacg gtatcataga tacattaaaa atgttttccg gagcatttgg ctttccttcc
7081 attctatgat tgtttccata ccgttgcgta tcactttcat aatctgctaa aaatgattta
7141 aagtcagact tacactcagt ccaaaggctg gaaaatgttt cagtatcatt gtgaaatatt
7201 gtatagcttg gtatcatctc atcatatatc cccaattcac catcttgatt gattgccgtc
7261 ctaaactctg aatggcggtt tacaatcatt gcaatataat aaagcattgc aggatatagt
7321 ttcattccct tttcctttat ttgtgtgata tccactttaa cggtcatgct gtaggtacaa
7381 ggtacacttg caaagtagtg gtcaaaatac tcttttctgt tccaactatt tttatcaatt
7441 ttttcaaata ccatctaagt tccctctcaa attcaagttt atcgctctaa tgaacaaaga
7501 tattatacca catttttgtg aatttttcaa cttgcccact tcgactgcac tcccgactta
7561 ataacttctt gaacacttgc cgaaaaagaa aaactgccgg gtacgtaccc ggggatcgat
7621 ccccggccga gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
7681 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
7741 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
7801 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
7861 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctt
7921 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
7981 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
8041 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
8101 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
8161 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
8221 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
8281 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
8341 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
8401 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
8461 gttaagggat tttggtcatg aaatgcaagt ttctaactaa catacatcat agttactaaa
8521 ctatggtaac tataatttta ttaactatat taagcactga ttagtactat aactcaatat
8581 aagcatatcc cctgtatcgt aactagagaa ccaaacgacg gaaaaagcga tatagataag
8641 tttagatagt gcaccaattt tttacttttc tattttagaa tctagcattt ccaatgctta
8701 ctaatactat ttttgatttt taacttgttt taaaaaagca cctactaatt aggattctag
8761 ctctatattc tagttataca ttctctatat tctagctcta tattctagtt atacattctc
8821 tatattctag ctctatattc tagttataca ttctctatat tctagctcta tattctagtt
8881 atacattctc tatattctag ctctatattc tagttataca ttctctatat tctagctcta
8941 tattctagtt atacattctc tatattctag ctctatattc tagttataca ttctctaaaa
9001 aacatatata tatatactac atatatatgt acaattattt taccaaaaga aatttttaaa
9061 acggcttaat ttcaatactt tgatgccatt ttttttttgc aattctcgta caattatttt
9121 accaaaagaa atttttatta attagttttt ttgcataaaa ataggactag aaaatcaatt
9181 ctaatcctat caaaaatcta cttgtttaat ctatatgtat agtacataaa t
//
