| 质粒名称: | pBEST-p15A-PLtetO1-tetR-T500 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 45391 |
| 存储实验室: | Vincent Noireaux |
|
载体骨架基本信息 |
|
| 载体名称: | pBEST-Luc |
| 载体出品公司: | Promega |
| 空载体大小: | 4486bp |
| 完整质粒大小: | 4486bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | tet repressor |
| 物种来源: | e. coli |
| 插入大小: | 623 bp |
| GenBank编号 | BAG71042.1 |
| 启动子: | PL-tetO1 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | sphI (未破坏) |
| 3’克隆位点: | NheI (未破坏) |
|
文献引用方法 |
|
| 材料和方法部分: | pBEST-p15A-PLtetO1-tetR-T500 was a gift from Vincent Noireaux (Addgene plasmid # 45391 ; http://n2t.net/addgene:45391 ; RRID:Addgene_45391) |
| 参考文献部分: | Efficient cell-free expression with the endogenous E. Coli RNA polymerase and sigma factor 70. Shin J, Noireaux V. J Biol Eng. 2010 Jun 24;4:8. 10.1186/1754-1611-4-8 PubMed 20576148 |
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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 pBEST-p15A-PLtetO1-tetR-T500 3240 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBEST-p15A-PLtetO1-tetR-T500
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3240)
AUTHORS Shin J, Noireaux V
TITLE Efficient cell-free expression with the endogenous E. Coli RNA
polymerase and sigma factor 70.
JOURNAL J Biol Eng. 2010 Jun 24;4:8.
PUBMED 20576148
REFERENCE 2 (bases 1 to 3240)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3240
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 1..19
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 26..44
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 68..90
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 97..720
/codon_start=1
/gene="tetR from transposon Tn10"
/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="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind complement(170..189)
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
rep_origin 1007..1552
/direction=RIGHT
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin.
"
primer_bind 1453..1472
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1669..1686
/label=L4440
/note="L4440 vector, forward primer"
CDS complement(1919..2779)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 2542..2561
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2780..2884)
/gene="bla"
/label=AmpR promoter
primer_bind 2952..2970
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tccctatcag tgatagagat tgacatccct atcagtgata gagatactga gcacagctag
61 caataatttt gtttaacttt aagaaggaga tataccatgt ctagattaga taaaagtaaa
121 gtgattaaca gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt aacaacccgt
181 aaactcgccc agaagctagg tgtagagcag cctacattgt attggcatgt aaaaaataag
241 cgggctttgc tcgacgcctt agccattgag atgttagata ggcaccatac tcacttttgc
301 cctttagaag gggaaagctg gcaagatttt ttacgtaata acgctaaaag ttttagatgt
361 gctttactaa gtcatcgcga tggagcaaaa gtacatttag gtacacggcc tacagaaaaa
421 cagtatgaaa ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt ttcactagag
481 aatgcattat atgcactcag cgctgtgggg cattttactt taggttgcgt attggaagat
541 caagagcatc aagtcgctaa agaagaaagg gaaacaccta ctactgatag tatgccgcca
601 ttattacgac aagctatcga attatttgat caccaaggtg cagagccagc cttcttattc
661 ggccttgaat tgatcatatg cggattagaa aaacaactta aatgtgaaag tgggtcttaa
721 ctcgagcaaa gcccgccgaa aggcgggctt ttctgtgtcg accgatgccc ttgagagcct
781 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
841 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc atgataagct gtcaaacatg
901 agaattgcaa cttatatcgt atggggctga cttcaggtgc tacatttgaa gagataaatt
961 gcactgaaat ctagaaatat tttatctgat taataagatg atcttcttga gatcgttttg
1021 gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc cttgcagggc ggtttttcga
1081 aggttctctg agctaccaac tctttgaacc gaggtaactg gcttggagga gcgcagtcac
1141 caaaacttgt cctttcagtt tagccttaac cggcgcatga cttcaagact aactcctcta
1201 aatcaattac cagtggctgc tgccagtggt gcttttgcat gtctttccgg gttggactca
1261 agacgatagt taccggataa ggcgcagcgg tcggactgaa cggggggttc gtgcatacag
1321 tccagcttgg agcgaactgc ctacccggaa ctgagtgtca ggcgtggaat gagacaaacg
1381 cggccataac agcggaatga caccggtaaa ccgaaaggca ggaacaggag agcgcacgag
1441 ggagccgcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccaccactg
1501 atttgagcgt cagatttcgt gatgcttgtc aggggggcgg agcctatgga aaaacggctt
1561 tgccgcggcc ctctcacttc cctgttaagt atcttcctgg catcttccag gaaatctccg
1621 ccccgttcgt aagccatttc cgctcgccgc agtcgaacga ccgagcgtag cgagtcagtg
1681 agcgaggaag cggaatatat cctgtatcac atattctgct gacgcaccgg tgcagccttt
1741 tttctcctgc cacatgaagc acttcactga caccctcatc agtgccaaca tagtaagcca
1801 gtatacactc cgctagggtc atgagattat caaaaaggat cttcacctag atccttttaa
1861 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
1921 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
1981 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
2041 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
2101 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
2161 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
2221 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
2281 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
2341 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
2401 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
2461 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
2521 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
2581 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
2641 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
2701 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
2761 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
2821 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
2881 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
2941 taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaattc tggcgaatcc
3001 tctgaccagc cagaaaacga cctttctgtg gtgaaaccgg atgctgcaat tcagagcgcc
3061 agcaagtggg ggacagcaga agacctgacc gccgcagagt ggatgtttga catggtgaag
3121 actatcgcac catcagccag aaaaccgaat tttgctgggt gggctaacga tatccgcctg
3181 atgcgtgaac gtgacggacg taaccaccgc gacatgtgtg tgctgttccg ctgggcatgc
//
