| 质粒名称: | pTriEx-mCherry-LOV2 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 81041 |
| 存储实验室: | Klaus Hahn |
|
载体骨架基本信息 |
|
| 载体名称: | pTriEx |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5040 bp |
| 完整质粒大小: | 6000 bp |
| 载体类型: | 哺乳动物表达，细菌表达，昆虫表达 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | LOV2 |
| 物种来源: | Synthetic |
| 插入大小: | 432 bp |
| 标签/融合蛋白: | mCherry（插入件上的N端子） |
| 5' 测序引物 | aagtatcgggccctttgtgc |
| 3' 测序引物 | GGCAGCCTGCACCTGAGGTTAATCAC |
|
文献引用方法 |
|
| 材料和方法部分: | pTriEx-mCherry-LOV2 was a gift from Klaus Hahn (Addgene plasmid # 81041 ; http://n2t.net/addgene:81041 ; RRID:Addgene_81041) |
| 参考文献部分: | LOVTRAP: an optogenetic system for photoinduced protein dissociation. Wang H, Vilela M, Winkler A, Tarnawski M, Schlichting I, Yumerefendi H, Kuhlman B, Liu R, Danuser G, Hahn KM. Nat Methods. 2016 Jul 18. doi: 10.1038/nmeth.3926. 10.1038/nmeth.3926 PubMed 27427858 |
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 pTriEx-mCherry-LOV2 6088 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION mammalian expression for wt LOV2.
ACCESSION .
VERSION .
KEYWORDS pTriEx-mCherry-LOV2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6088)
AUTHORS Wang H, Vilela M, Winkler A, Tarnawski M, Schlichting I, Yumerefendi
H, Kuhlman B, Liu R, Danuser G, Hahn KM
TITLE LOVTRAP: an optogenetic system for photoinduced protein
dissociation.
JOURNAL Nat Methods. 2016 Jul 18. doi: 10.1038/nmeth.3926.
PUBMED 27427858
REFERENCE 2 (bases 1 to 6088)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6088
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_recomb 110..937
/label=baculovirus recombination region (lef2/ORF603)
/note="contains ORF603 and part of lef2"
CDS 110..234
/codon_start=3
/product="baculovirus late expression factor 2"
/label=lef2
/translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
CDS complement(272..877)
/codon_start=1
/product="baculovirus ORF603 protein"
/label=ORF603
/translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
enhancer 1005..1308
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1310..1509
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1463..1483
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 1715..1737
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1799..1818
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
primer_bind 1860..1879
/label=T7
/note="T7 promoter, forward primer"
promoter 1860..1878
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1879..1903
/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)."
promoter 1919..2028
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
CDS 2043..2750
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(2188..2206)
/label=mCherry-R
/note="mCherry, reverse primer"
CDS 3231..3263
/codon_start=1
/product="HSV (herpes simplex virus) epitope tag"
/label=HSV tag
/translation="QPELAPEDPED"
CDS 3270..3293
/codon_start=1
/product="8xHis affinity tag"
/label=8xHis
/translation="HHHHHHHH"
primer_bind complement(3383..3402)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_site 3448..3503
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind complement(3502..3521)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
terminator 3591..3638
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
misc_recomb 3651..4356
/label=baculovirus recombination region (ORF1629)
/note="contains part of ORF1629"
3'UTR 3739..4111
/label=polyhedrin 3' UTR
/note="3' UTR from the Autographa californica
nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
CDS complement(3768..4356)
/codon_start=2
/product="baculovirus capsid-associated protein"
/label=ORF1629
/note="required for viral replication"
/translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
protein_bind 4365..4398
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
rep_origin complement(4566..5153)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4645..4664)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5166..6026)
/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 5789..5808
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
ORIGIN
1 aactgttaca ttgcaaacgt ggtttcgtgt gccaagtgtg aaaaccgatg tttaatcaag
61 gctctgacgc atttctacaa ccacgactcc aagtgtgtgg gtgaagtcat gcatctttta
121 atcaaatccc aagatgtgta taaaccacca aactgccaaa aaatgaaaac tgtcgacaag
181 ctctgtccgt ttgctggcaa ctgcaagggt ctcaatccta tttgtaatta ttgaataata
241 aaacaattat aaatgtcaaa tttgtttttt attaacgata caaaccaaac gcaacaagaa
301 catttgtagt attatctata attgaaaacg cgtagttata atcgctgagg taatatttaa
361 aatcattttc aaatgattca cagttaattt gcgacaatat aattttattt tcacataaac
421 tagacgcctt gtcgtcttct tcttcgtatt ccttctcttt ttcatttttc tcttcataaa
481 aattaacata gttattatcg tatccatata tgtatctatc gtatagagta aattttttgt
541 tgtcataaat atatatgtct tttttaatgg ggtgtatagt accgctgcgc atagtttttc
601 tgtaatttac aacagtgcta ttttctggta gttcttcgga gtgtgttgct ttaattatta
661 aatttatata atcaatgaat ttgggatcgt cggttttgta caatatgttg ccggcatagt
721 acgcagcttc ttctagttca attacaccat tttttagcag caccggatta acataacttt
781 ccaaaatgtt gtacgaaccg ttaaacaaaa acagttcacc tcccttttct atactattgt
841 ctgcgagcag ttgtttgttg ttaaaaataa cagccattgt aatgagacgc acaaactaat
901 atcacaaact ggaaatgtct atcaatatat agttgctgat ggccggccta ttaatagtaa
961 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
1021 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
1081 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
1141 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt
1201 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac
1261 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatgct gatgcggttt
1321 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
1381 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
1441 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
1501 ataagcagac gtcgtttagt gaaccgtcag atcactagat gctttattgc ggtagtttat
1561 cacagttaaa ttgctaacgc cagtctcgaa cttaacgtgc agaagttggt cgtgaggcac
1621 tgggcaggta agtatcgggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
1681 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
1741 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
1801 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggatcg gaccgaaatt
1861 aatacgactc actatagggg aattgtgagc ggataacaat tccccggagt taatccggga
1921 cctttaattc aacccaacac aatatattat agttaaataa gaattattat caaatcattt
1981 gtatattaat taaaatacta tactgtaaat tacattttat ttacaatcaa aggagatata
2041 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
2101 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
2161 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
2221 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
2281 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
2341 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
2401 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccctg
2461 taatgcagaa gaagactatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
2521 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
2581 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
2641 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
2701 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag ggttctggat
2761 ccttggctac tacacttgaa cgtattgaga agaactttgt cattactgac ccaagattgc
2821 cagataatcc cattatattc gcgtccgata gtttcttgca gttgacagaa tatagccgtg
2881 aagaaatttt gggaagaaac tgcaggtttc tacaaggtcc tgaaactgat cgcgcgacag
2941 tgagaaaaat tagagatgcc atagataacc aaacagaggt cactgttcag ctgattaatt
3001 atacaaagag tggtaaaaag ttctggaacc tctttcactt gcagcctatg cgagatcaga
3061 agggagatgt ccagtacttt attggggttc agttggatgg aactgagcat gtccgagatg
3121 ctgccgagag agagggagtc atgctgatta agaaaactgc agaaaatatt gatgaggcgg
3181 caaaagaact ttaaaagctt gcggccgcac agctgtatac acgtgcaagc cagccagaac
3241 tcgccccgga agaccccgag gatctcgagc accaccatca ccatcaccat cactaagtga
3301 ttaacctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
3361 tcacaaatac cactgagatc gatctttttc cctctgccaa aaattatggg gacatcatga
3421 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
3481 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
3541 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgtaa ctagcataac
3601 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaagcatgc ggaggaaatt
3661 ctccttgaag tttccctggt gttcaaagta aaggagtttg caccagacgc acctctgttc
3721 actggtccgg cgtattaaaa cacgatacat tgttattagt acatttatta agcgctagat
3781 tctgtgcgtt gttgatttac agacaattgt tgtacgtatt ttaataattc attaaattta
3841 taatctttag ggtggtatgt tagagcgaaa atcaaatgat tttcagcgtc tttatatctg
3901 aatttaaata ttaaatcctc aatagatttg taaaataggt ttcgattagt ttcaaacaag
3961 ggttgttttt ccgaaccgat ggctggacta tctaatggat tttcgctcaa cgccacaaaa
4021 cttgccaaat cttgtagcag caatctagct ttgtcgatat tcgtttgtgt tttgttttgt
4081 aataaaggtt cgacgtcgtt caaaatatta tgcgcttttg tatttctttc atcactgtcg
4141 ttagtgtaca attgactcga cgtaaacacg ttaaatagag cttggacata tttaacatcg
4201 ggcgtgttag ctttattagg ccgattatcg tcgtcgtccc aaccctcgtc gttagaagtt
4261 gcttccgaag acgattttgc catagccaca cgacgcctat taattgtgtc ggctaacacg
4321 tccgcgatca aatttgtagt tgagcttttt ggaattgcga tcgcataact tcgtatagca
4381 tacattatac gaagttataa gctcggaacg ctgcgctcgg tcgttcggct gcggcgagcg
4441 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
4501 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
4561 gcgtttttcc ataggctccg ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
4621 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
4681 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
4741 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
4801 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
4861 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
4921 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
4981 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
5041 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
5101 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
5161 ctttgttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
5221 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
5281 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
5341 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
5401 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
5461 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
5521 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
5581 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
5641 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
5701 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
5761 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
5821 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
5881 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
5941 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
6001 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
6061 cagggttatt gtctcatgtc cgcgcgtt
//
