| 质粒名称: | pRecT_2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 167546 |
| 存储实验室: | Howard Hang |
|
载体骨架基本信息 |
|
| 载体名称: | plZ12-Spec |
| 载体出品公司: | Luciano Marraffini |
| 空载体大小: | 3729bp |
| 完整质粒大小: | 6411 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 大观霉素，50μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | 大肠杆菌在IPTG诱导型启动子下的Com15-recT，具有lacI |
| 物种来源: | -- |
| 插入大小: | 2896 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | IPTG |
| 标签/融合蛋白: | Gibson Cloning |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | TTATTATAACATGTATTCACGAACG |
| 3’测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pRecT_2 was a gift from Howard Hang (Addgene plasmid # 167546 ; http://n2t.net/addgene:167546 ; RRID:Addgene_167546) |
| 参考文献部分: | RecT recombinase expression enables efficient gene editing in Enterococcus. Chen V, Griffin ME, Maguin P, Varble A, Hang HC. Appl Environ Microbiol. 2021 Jul 7:AEM0084421. doi: 10.1128/AEM.00844-21. 10.1128/AEM.00844-21 PubMed 34232061 |
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
载体简介
载体序列
LOCUS pRecT_2 6411 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Encodes an IPTG inducible recT from E. faecium Com15 used to
ectopically express recT in Enterococcus for recombineering..
ACCESSION .
VERSION .
KEYWORDS pRecT_2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6411)
TITLE RecT recombinase expression enables efficient gene editing in
Enterococcus
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 6411)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6411
/organism="synthetic DNA construct"
/mol_type="other DNA"
terminator 2246..2332
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 2351..2378
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 2458..2492
/label=lambda t0 terminator
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
protein_bind 2596..2612
/label=lac operator
/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)."
CDS 3862..4944
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKTAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind complement(3881..3900)
/label=LacI-R
/note="LacI, reverse primer"
CDS complement(5922..5933)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
ORIGIN
1 aaaatctgcg gttgcggcgt tagctataga agaatatgca agaaaggaat cagaacaaaa
61 aaaataagcg aaagctcgcg tttttagaag gatacgagtt ttcgctactt gtttttgata
121 aggtaattat atcatggcta ttaaaaatac taaagctaga aattttggat ttttattata
181 tcctgactca attcctaatg attggaaaga aaaattagag agtttgggcg tatctatggc
241 tgtcagtcct ttacacgata tggacgaaaa aaaagataaa gatacatgga atagtagtga
301 tgttatacga aatggaaagc actataaaaa accacactat cacgttatat atattgcacg
361 aaatcctgta acaatagaaa gcgttaggaa caagattaag cgaaaattgg ggaatagttc
421 agttgctcat gttgagatac ttgattatat caaaggttca tatgaatatt tgactcatga
481 atcaaaggac gctattgcta agaataaaca tatatacgac aaaaaagata ttttgaacat
541 taatgatttt gatattgacc gctatataac acttgatgaa agccaaaaaa gaaaattgaa
601 gaatttactt ttagatatag tggatgacta taatttggta aatacaaaag atttaatggc
661 ttttattcgc cttaggggag cggagtttgg aattttaaat acgaatgatg taaaagatat
721 tgtttcaaca aactctagcg cctttagatt atggtttgag ggcaattatc agtgtggata
781 tagagcaagt tatgcaaagg ttcttgatgc tgaaacgggg gaaataaaat gacaaacaaa
841 gaaaaagagt tatttgctga aaatgaggaa ttaaaaaaag aaattaagga cttaaaagag
901 cgtattgaaa gatacagaga aatggaagtt gaattaagta caacaataga tttattgaga
961 ggagggatta ttgaataaat aaaagccccc ctgacgaaag tccaagggtt tattgttttc
1021 taaaatctga ttaccaatta gaatgaatat ttcccaaata ttaaataata aaacaaaaaa
1081 attaaaaaag tgtttccacc attttttcaa tttttttata atttttttaa tctgttattt
1141 aaatagttta tagttaaatt tacattttca ttagtccatt caatattctc tccaagataa
1201 ctacgaactg ctaacaaaat tctctcccta tgttctaatg gagaagattc agccactgca
1261 tttcccgcaa tatcttttgg tatgatttta cccgtgtcca tagttaaaat catacggcat
1321 aaagttaata tagagttggt ttcatcatcc tgataattat ctattaattc ctctgacgaa
1381 tccataatgg ctcttctcac atcagaaaat ggaatatcag gtagtaattc ctctaagtca
1441 taatttccgt atattctttt attttttcgt tttgcttggt aaagcattat ggttaaatct
1501 gaatttaatt ccttctgagg aatgtatcct tgttcataaa gctcttgtaa ccattctcca
1561 taaataaatt cttgtttggg aggatgattc cacggtacca tttcttgctg aataataatt
1621 gttaattcaa tatatcgtaa gttgctttta tctcctattt tttttgaaat aggtctaatt
1681 ttttgtataa gtatttcttt actttgatct gtcaatggtt cagatacgac gactaaaaag
1741 tcaagatcac tatttggttt tagtccactc tcaactcctg atccaaacat gtaagtacca
1801 ataaggttat tttttaaatg tttccgaagt atttttttca ctttattaat ttgttcgtat
1861 gtattcaaat atatcctcct cactattttg attagtacct attttatatc catagttgtt
1921 aattaaataa acttaattta gtttatttat agatttcatt ggcttctaaa ttttttatct
1981 agataataat tattttagtt aattttattc tagattatat atgatatgat ctttcatttc
2041 cataaaacta aagtaagtgt aaacctattc attgttttaa aaatatctct tgccagtcac
2101 gttacgttat tagttatagt tattataaca tgtattcacg aacgaaaatc gacctgcagg
2161 catgcaagct tggcgttata aaattagtat aattatagca cgagctctga taaatatgaa
2221 catgcgactc tctagcttga ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
2281 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccgct
2341 ctagctaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttgtt
2401 aactctagag ctgcctgccg cgtttcggtg atgaagatct tcccgatgat taattaattc
2461 agaacgctcg gttgccgccg ggcgtttttt atgcagcaat ggcaagaacg ttgctcgagg
2521 gtaaatgtga gcactcacaa ttcattttgc aaaagttgtt gactttatct acaaggtgtg
2581 gcataatgtg tggaattgtg agcggataac aattaagctt gtacttagga ggatgattat
2641 ttatggcaac aaatgaatcg ttaaaaaacc aattggcaga aaagccacag aaacaagttg
2701 caccaggaca gttagggctt aaagctctaa tgaatacacc aacaatgaga aagaaatttg
2761 aagaagtact tcatgacaat gctaatgctt ttatgtcgaa tgttatgact cttgtatcta
2821 atgacaatta tcttgcagat agtgaaccga tgtctatcat gagtggtgcg ttaactgctg
2881 caacattaaa tcttggacta gataagaatc ttggctatgc ctatctcgtt ccatttaata
2941 gcaaaaacaa gcaaacagga aaatgggaaa agaaagctca atttatgctt ggctataaag
3001 gatatatcca attagcccaa cgatcaggta aatacaaagc attaaatgtg attgaagttt
3061 acgaaggaga actaaaaagc tggaaccgac tgacagaaga gttcgagttt gatccaaatg
3121 gtagaacatc tgatgaagtc attggatatg ttggctattt tgaattactg aatggattca
3181 agaaaactgt ctattggacc aaacaagaaa ttgaagctca tcgaattggt aacaataaag
3241 atcgagataa gacaaagtta agtggtgtgt gggcatctga ttacaatgca atggcacgaa
3301 aaactgtttt gagaaatctt ctttctaaat gggggatctt atccattgaa atgcaagaag
3361 ctaccacatc ggatgagaga gtccaaagag ttcaagaaga tggcagcatt attgctgaaa
3421 cagaagttga ggaagatatt cctgaaagaa aagaagcaga aaaagtagaa ataaacgctg
3481 acaatggatc aacagctttt gaagagtatg cagaatcagt tcagacagat gaagcacaag
3541 aatcgctgtt cgatgcatcg aatccaccat taaataaata gcccgggtaa ctagtaactg
3601 cagatgctag ctcgcatgca agctaattcg gtggaaacga ggtcatcatt tccttccgaa
3661 aaaacggttg catttaaatc ttacatatgt aatactttca aagactacat ttgtaagatt
3721 tgatgtttga gtcggctgaa agatcgtacg taccaattat tgtttcgtga ttgttcaagc
3781 cataacactg tagggatagt ggaaagagtg cttcatctgg ttacgatcaa tcaaatattc
3841 aaacggaggg agacgatttt gatgaaacca gtaacgttat acgatgtcgc agagtatgcc
3901 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
3961 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
4021 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
4081 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
4141 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaaacggc ggtgcacaat
4201 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
4261 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
4321 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
4381 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
4441 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
4501 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
4561 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
4621 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
4681 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
4741 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
4801 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
4861 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
4921 tcccgactgg aaagcgggca gtgaatgagt gatcgttaaa tttatactgc aatcggatgc
4981 gattattgaa taaaagatat gagagattta tctaatttct tttttcttgt aaaaaaagaa
5041 agttcttaaa ggttttatag ttttggtcgt aggaagatct ggatcctaat gaattcatct
5101 gcaggggccc atcagtctga cgaccaagag agccataaac accaatagcc ttaacatcat
5161 ccccatattt atccaatatt cgttccttaa tttcatgaac aatcttcatt ctttcttctc
5221 tagtcattat tattggtcca ttcactattc tcattccctt ttcagataat tttagatttg
5281 cttttctaaa taagaatatt tggagagcac cgttcttatt cagctattaa taactcgtct
5341 tcctaagcat ccttcaatcc ttttaataac aattatagca tctaatcttc aacaaactgg
5401 cccgtttgtt gaactactct ttaataaaat aatttttccg ttcccaattc cacattgcaa
5461 taatagaaaa tccatcttca tcggcttttt cgtcatcatc tgtatgaatc aaatcgcctt
5521 cttctgtgtc atcaaggttt aattttttat gtatttcttt taacaaacca ccataggaga
5581 ttaacctttt acggtgtaaa ccttcctcca aatcagacaa acgtttcaaa ttcttttctt
5641 catcatcggt cataaaatcc gtatccttta caggatattt tgcagtttcg tcaattgccg
5701 attgtatatc cgatttatat ttatttttcg gtcgattttt tattaaaacg tctcaaaatc
5761 gtttctgaga cgttttagcg tttatttcgt ttagttatcg gcataatcgt taaaacaggc
5821 gttatcgtag cgtaaaagcc cttgagcgta gcgtggcttt gcagcgaaga tgttgtctgt
5881 tagattatga aagccgatga ctgaatgaaa taataagcgc agcgtccttc tatttcggtt
5941 ggaggaggct caagggagtt tgagggaatg aaattccctc atgggtttga ttttaaaaat
6001 tgcttgcaat tttgccgagc ggtagcgctg gaaaattttt gaaaaaaatt tggaatttgg
6061 aaaaaaatgg ggggaaagga agcgaatttt gcttccgtac tacgaccccc cattaagtgc
6121 cgagtgccaa tttttgtgcc aaaaacgctc tatcccaact ggctcaaggg tttgaggggt
6181 ttttcaatcg ccaacgaatc gccaacgttt tcgccaacgt tttttataaa tctatattta
6241 agtagcttta ttgttgtttt tatgattaca aagtgataca ctaattttat aaaattattt
6301 gattggagtt ttttaaatgg tgatttcaga atcgaaaaaa agagttatga tttctctgac
6361 aaaagagcaa gataaaaaat taacagatat ggcgaaacaa aaaggttttt c
//
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