| 质粒名称: | pSC101_TIMER |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 103057 |
| 存储实验室: | Dirk Bumann |
|
载体骨架基本信息 |
|
| 载体名称: | pSC101 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 9262 bp |
| 完整质粒大小: | -- |
| 载体修饰: | 用pSC101复制子在pUA66中克隆了PybaJ timerbac |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | TIMER-bac |
| 物种来源: | Synthetic; Discosoma spp |
| 插入大小: | -- |
| 基因突变: | mutations: S4T and S179T |
| GenBank编号 | -- |
| 启动子 | constitutive ybaJp promoter |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 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: | pSC101_TIMER was a gift from Dirk Bumann (Addgene plasmid # 103057 ; http://n2t.net/addgene:103057 ; RRID:Addgene_103057) |
| 参考文献部分: | Phenotypic variation of Salmonella in host tissues delays eradication by antimicrobial chemotherapy. Claudi B, Sprote P, Chirkova A, Personnic N, Zankl J, Schurmann N, Schmidt A, Bumann D. Cell. 2014 Aug 14;158(4):722-733. doi: 10.1016/j.cell.2014.06.045. 10.1016/j.cell.2014.06.045 PubMed 25126781 |
买家导航
pSC101TIMER载体基本信息
pSC101TIMER载体图谱质粒图谱和多克隆位点信息
pSC101TIMER载体简介
pSC101TIMER载体序列
LOCUS pSC101_TIMER 4946 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION low copy plasmid for expression of a variant of the fluorescent
timer protein suitable for bacteria under the control of the
constitutive ybaJp promoter (works in E. coli and Salmonella
enterica).
ACCESSION .
VERSION .
KEYWORDS pSC101_TIMER
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4946)
AUTHORS Claudi B, Sprote P, Chirkova A, Personnic N, Zankl J, Schurmann N,
Schmidt A, Bumann D
TITLE Phenotypic variation of Salmonella in host tissues delays
eradication by antimicrobial chemotherapy.
JOURNAL Cell. 2014 Aug 14;158(4):722-733. doi: 10.1016/j.cell.2014.06.045.
PUBMED 25126781
REFERENCE 2 (bases 1 to 4946)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4946
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 64..158
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
rep_origin 650..872
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
CDS 920..1870
/codon_start=1
/gene="rep101"
/product="RepA protein needed for replication with the
pSC101 origin"
/label=Rep101
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
terminator 2408..2494
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 2622..2641
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(3416..3434)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind complement(3466..3486)
/label=DsRed1-N
/note="DsRed1, reverse primer"
primer_bind 3684..3703
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 3878..3901
/label=DsRed1-C
/note="DsRed1, forward primer"
primer_bind complement(3988..4006)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
CDS 4185..33
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4239..4258)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4849..4868
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
ORIGIN
1 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gagagctcgc
61 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
121 cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcactagg gacagtaaga
181 cgggtaagcc tgttgatgat accgctgcct tactgggtgc attagccagt ctgaatgacc
241 tgtcacggga taatccgaag tggtcagact ggaaaatcag agggcaggaa ctgctgaaca
301 gcaaaaagtc agatagcacc acatagcaga cccgccataa aacgccctga gaagcccgtg
361 acgggctttt cttgtattat gggtagtttc cttgcatgaa tccataaaag gcgcctgtag
421 tgccatttac ccccattcac tgccagagcc gtgagcgcag cgaactgaat gtcacgaaaa
481 agacagcgac tcaggtgcct gatggtcgga gacaaaagga atattcagcg atttgcccga
541 gcttgcgagg gtgctactta agcctttagg gttttaaggt ctgttttgta gaggagcaaa
601 cagcgtttgc gacatccttt tgtaatactg cggaactgac taaagtagtg agttatacac
661 agggctggga tctattcttt ttatcttttt ttattctttc tttattctat aaattataac
721 cacttgaata taaacaaaaa aaacacacaa aggtctagcg gaatttacag agggtctagc
781 agaatttaca agttttccag caaaggtcta gcagaattta cagataccca caactcaaag
841 gaaaaggact agtaattatc attgactagc ccatctcaat tggtatagtg attaaaatca
901 cctagaccaa ttgagatgta tgtctgaatt agttgttttc aaagcaaatg aactagcgat
961 tagtcgctat gacttaacgg agcatgaaac caagctaatt ttatgctgtg tggcactact
1021 caaccccacg attgaaaacc ctacaaggaa agaacggacg gtatcgttca cttataacca
1081 atacgctcag atgatgaaca tcagtaggga aaatgcttat ggtgtattag ctaaagcaac
1141 cagagagctg atgacgagaa ctgtggaaat caggaatcct ttggttaaag gctttgagat
1201 tttccagtgg acaaactatg ccaagttctc aagcgaaaaa ttagaattag tttttagtga
1261 agagatattg ccttatcttt tccagttaaa aaaattcata aaatataatc tggaacatgt
1321 taagtctttt gaaaacaaat actctatgag gatttatgag tggttattaa aagaactaac
1381 acaaaagaaa actcacaagg caaatataga gattagcctt gatgaattta agttcatgtt
1441 aatgcttgaa aataactacc atgagtttaa aaggcttaac caatgggttt tgaaaccaat
1501 aagtaaagat ttaaacactt acagcaatat gaaattggtg gttgataagc gaggccgccc
1561 gactgatacg ttgattttcc aagttgaact agatagacaa atggatctcg taaccgaact
1621 tgagaacaac cagataaaaa tgaatggtga caaaatacca acaaccatta catcagattc
1681 ctacctacat aacggactaa gaaaaacact acacgatgct ttaactgcaa aaattcagct
1741 caccagtttt gaggcaaaat ttttgagtga catgcaaagt aagtatgatc tcaatggttc
1801 gttctcatgg ctcacgcaaa aacaacgaac cacactagag aacatactgg ctaaatacgg
1861 aaggatctga ggttcttatg gctcttgtat ctatcagtga agcatcaaga ctaacaaaca
1921 aaagtagaac aactgttcac cgttacatat caaagggaaa actgtccata tgcacagatg
1981 aaaacggtgt aaaaaagata gatacatcag agcttttacg agtttttggt gcattcaaag
2041 ctgttcacca tgaacagatc gacaatgtaa cagatgaaca gcatgtaaca cctaatagaa
2101 caggtgaaac cagtaaaaca aagcaactag aacatgaaat tgaacacctg agacaacttg
2161 ttacagctca acagtcacac atagacagcc tgaaacaggc gatgctgctt atcgaatcaa
2221 agctgccgac aacacgggag ccagtgacgc ctcccgtggg gaaaaaatca tggcaattct
2281 ggaagaaata gcgctttcag ccggcaaacc ggctgaagcc ggatctgcga ttctgataac
2341 aaactagcaa caccagaaca gcccgtttgc gggcagcaaa acccgtaccc taggtctagg
2401 gcggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
2461 cccagtcttt cgactgagcc tttcgtttta tttgatgcct ctagagcttg catgcccaag
2521 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
2581 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
2641 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
2701 atcgtataat ccttactgcg ttaaaggctt ttcttaggaa agttggccat ttcttaattc
2761 agccattaat taagaaatat taagaatatt cctggctatt ttctcctgtc agagtctatt
2821 gttttagcct gaaaagctaa aaaacgttaa cccaatgatt acacaaacaa taaaactggt
2881 tcctttttag gcgaccgacg atcactgtta aaattcgaaa aagtatggca acacgcggct
2941 ttcacgcaat tgtaattttt agtaatatga cgatgaaaag ttttttagag tagattatag
3001 ttaaatcata aggtgacgtg ggaagtacca ggttagttag ttgtatccat cccgaaggtg
3061 ttcggttagt ttaagccact aaaaagggga tgcattatgg acgaatactc acccaaaaga
3121 catgatatag cacagcttaa atttctttgc gaaaccctgt atcatgactg tcttgcaaac
3181 cttgaagaaa gcaaccatgg ctgggttaat gacccgacct cagcagtcaa cctgcagctc
3241 aatgagctga tagagatcct ctagatttaa gaaggagata tacatatggc atccaccgag
3301 gacgtcatca aggagttcat gcgcttcaaa gtgcgcatgg agggctccgt gaacggccac
3361 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
3421 ctgaaggtga ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc cccccagttc
3481 cagtacggct ccaaggtgta cgtgaagcac cccgccgaca tccccgacta caagaagctg
3541 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
3601 accgtgaccc aggactcctc cctgcaggac ggctgcttca tctacaaggt gaagttcatc
3661 ggcgtgaact tcccctccga cggccccgta atgcagaaga agactatggg ctgggagccc
3721 tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg gcgagatcca caaggccctg
3781 aagctgaagg acggcggcca ctacctggtg gagttcaagt ccatctacat ggccaagaag
3841 cccgtgcaac tgcccggcta ctactacgtg gacaccaagc tggacatcac ctcccacaac
3901 gaggactaca ccatcgtgga gcagtacgaa cgcaccgagg gccgctacca cctgttcctg
3961 tagaagcttc gaggtgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
4021 tcatgataat aatggtttct tagacgtcgg aattgccagc tggggcgccc tctggtaagg
4081 ttgggaagcc ctgcaaagta aactggatgg ctttcttgcc gccaaggatc tgatggcgca
4141 ggggatcaag atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
4201 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
4261 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
4321 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
4381 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
4441 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
4501 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
4561 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
4621 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
4681 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
4741 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
4801 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
4861 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
4921 tcgccgctcc cgattcgcag cgcatc
//
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pSC101TIMER载体质粒应用举例
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