| 质粒名称: | pBEST-p15A-PLtetO1-deGFP-T500 |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 45392 |
| 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: | 4486 bp |
| 完整质粒大小: | 4486 bp |
| 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: | deGFP |
| 物种来源: | e. coli |
| 启动子: | PL-tetO1 |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 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-deGFP-T500 was a gift from Vincent Noireaux (Addgene plasmid # 45392 ; http://n2t.net/addgene:45392 ; RRID:Addgene_45392) |
| 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
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载体序列
LOCUS pBEST-p15A-PLtetO1-deGFP-T500 3293 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pBEST-p15A-PLtetO1-deGFP-T500
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3293)
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 3293)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..3293
/organism="synthetic DNA construct"
/mol_type="other DNA"
RBS 7..29
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
primer_bind complement(66..87)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(327..346)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 674..695
/label=EGFP-C
/note="EGFP, forward primer"
rep_origin 1000..1544
/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 1445..1464
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 1661..1678
/label=L4440
/note="L4440 vector, forward primer"
CDS complement(1911..2771)
/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 2534..2553
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(2772..2876)
/gene="bla"
/label=AmpR promoter
primer_bind 2944..2962
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
protein_bind 3233..3251
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 3258..3276
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
1 aataattttg tttaacttta agaaggagat ataccatgga gcttttcact ggcgttgttc
61 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
121 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
181 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
241 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
301 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
361 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
421 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
481 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
541 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
601 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
661 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc taactcgagc
721 aaagcccgcc gaaaggcggg cttttctgtg tcgaccgatg cccttgagag ccttcaaccc
781 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
841 ctttatcatg caactcgtag gacaggtgcc ggcatgataa gctgtcaaac atgagaatta
901 caacttatat cgtatggggc tgacttcagg tgctacattt gaagagataa attgcactga
961 aatctagaaa tattttatct gattaataag atgatcttct tgagatcgtt ttggtctgcg
1021 cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt cgaaggttct
1081 ctgagctacc aactctttga accgaggtaa ctggcttgga ggagcgcagt caccaaaact
1141 tgtcctttca gtttagcctt aaccggcgca tgacttcaag actaactcct ctaaatcaat
1201 taccagtggc tgctgccagt ggtgcttttg catgtctttc cgggttggac tcaagacgat
1261 agttaccgga taaggcgcag cggtcggact gaacgggggg ttcgtgcata cagtccagct
1321 tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa acgcggccat
1381 aacagcggaa tgacaccggt aaaccgaaag gcaggaacag gagagcgcac gagggagccg
1441 ccaggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac tgatttgagc
1501 gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc tttgccgcgg
1561 ccctctcact tccctgttaa gtatcttcct ggcatcttcc aggaaatctc cgccccgttc
1621 gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag tgagcgagga
1681 agcggaatat atcctgtatc acatattctg ctgacgcacc ggtgcagcct tttttctcct
1741 gccacatgaa gcacttcact gacaccctca tcagtgccaa catagtaagc cagtatacac
1801 tccgctaggg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
1861 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
1921 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
1981 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
2041 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
2101 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
2161 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
2221 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
2281 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
2341 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
2401 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
2461 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
2521 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
2581 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
2641 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
2701 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
2761 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
2821 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
2881 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
2941 aaaaataggc gtatcacgag gccctttcgt cttcaagaat tctggcgaat cctctgacca
3001 gccagaaaac gacctttctg tggtgaaacc ggatgctgca attcagagcg gcagcaagtg
3061 ggggacagca gaagacctga ccgccgcaga gtggatgttt gacatggtga agactatcgc
3121 accatcagcc agaaaaccga attttgctgg gtgggctaac gatatccgcc tgatgcgtga
3181 acgtgacgga cgtaaccacc gcgacatgtg tgtgctgttc cgctgggcat gctccctatc
3241 agtgatagag attgacatcc ctatcagtga tagagatact gagcacagct agc
//
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