| 质粒名称: | pTriEx-mCherry-PA-Rac1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 22027 |
| 存储实验室: | Klaus Hahn |
|
载体骨架基本信息 |
|
| 载体名称: | pTriEx |
| 载体出品公司: | Novagen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6000 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，细菌表达，昆虫表达 |
| 筛选标记:-- | |
| 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: | PA-Rac1,LOV(Leu404-Leu546)-Rac1(Ile4)-Q61L/E91H/N92H |
| 物种来源: | H. sapiens (human); Avena sativa (oat) |
| 插入大小: | 1100 bp |
| 基因突变: | Rac1从I4开始，包含突变Q61L、E91H和N92H（导致Rac1的光激活类似物） |
| GenBank编号 | AAA36544 |
| 启动子 | -- |
| 标签/融合蛋白: |
6X His（主干上的N终端）
mCherry（主干网上的N个终端）
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | Bam HI(未破坏) |
| 3' 克隆位点: | HindIII(未破坏) |
| 5' 测序引物 | pTriExUP |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pTriEx-mCherry-PA-Rac1 was a gift from Klaus Hahn (Addgene plasmid # 22027 ; http://n2t.net/addgene:22027 ; RRID:Addgene_22027) |
| 参考文献部分: | A genetically encoded photoactivatable Rac controls the motility of living cells. Wu YI, Frey D, Lungu OI, Jaehrig A, Schlichting I, Kuhlman B, Hahn KM. Nature. 2009 Sep 3. 461(7260):104-8. 10.1038/nature08241 PubMed 19693014 |
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-PA-Rac1 6747 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pTriEx-mCherry-PA-Rac1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6747)
AUTHORS Wu YI, Frey D, Lungu OI, Jaehrig A, Schlichting I, Kuhlman B, Hahn
KM
TITLE A genetically encoded photoactivatable Rac controls the motility of
living cells.
JOURNAL Nature. 2009 Sep 3. 461(7260):104-8.
PUBMED 19693014
REFERENCE 2 (bases 1 to 6747)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6747
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_recomb 1..706
/label=baculovirus recombination region (ORF1629)
/note="contains part of ORF1629"
CDS 1..589
/codon_start=2
/product="baculovirus capsid-associated protein"
/label=ORF1629
/note="required for viral replication"
/translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
3'UTR 246..618
/label=polyhedrin 3' UTR
/note="3' UTR from the Autographa californica
nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
terminator 719..766
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 836..855
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
polyA_site 854..909
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal"
primer_bind 955..974
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
CDS complement(1064..1087)
/codon_start=1
/product="8xHis affinity tag"
/label=8xHis
/translation="HHHHHHHH"
CDS complement(1094..1126)
/codon_start=1
/product="HSV (herpes simplex virus) epitope tag"
/label=HSV tag
/translation="QPELAPEDPED"
CDS complement(2168..2872)
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
primer_bind complement(2443..2462)
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 2712..2730
/label=mCherry-R
/note="mCherry, reverse primer"
CDS complement(2879..2896)
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
promoter complement(2917..3026)
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
protein_bind 3042..3066
/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)."
primer_bind complement(3066..3085)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3067..3085)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3127..3146)
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
primer_bind complement(3208..3230)
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
promoter complement(3436..3635)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(3462..3482)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 3637..3940
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
misc_recomb 4008..4835
/label=baculovirus recombination region (lef2/ORF603)
/note="contains ORF603 and part of lef2"
CDS 4068..4673
/codon_start=1
/product="baculovirus ORF603 protein"
/label=ORF603
/translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
CDS complement(4711..4835)
/codon_start=3
/product="baculovirus late expression factor 2"
/label=lef2
/translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
CDS 5078..5938
/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 complement(5296..5315)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 5951..6538
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6440..6459
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
protein_bind complement(6706..6739)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
ORIGIN
1 aattccaaaa agctcaacta caaatttgat cgcggacgtg ttagccgaca caattaatag
61 gcgtcgtgtg gctatggcaa aatcgtcttc ggaagcaact tctaacgacg agggttggga
121 cgacgacgat aatcggccta ataaagctaa cacgcccgat gttaaatatg tccaagctct
181 atttaacgtg tttacgtcga gtcaattgta cactaacgac agtgatgaaa gaaatacaaa
241 agcgcataat attttgaacg acgtcgaacc tttattacaa aacaaaacac aaacgaatat
301 cgacaaagct agattgctgc tacaagattt ggcaagtttt gtggcgttga gcgaaaatcc
361 attagatagt ccagccatcg gttcggaaaa acaacccttg tttgaaacta atcgaaacct
421 attttacaaa tctattgagg atttaatatt taaattcaga tataaagacg ctgaaaatca
481 tttgattttc gctctaacat accaccctaa agattataaa tttaatgaat tattaaaata
541 cgtacaacaa ttgtctgtaa atcaacaacg cacagaatct agcgcttaat aaatgtacta
601 ataacaatgt atcgtgtttt aatacgccgg accagtgaac agaggtgcgt ctggtgcaaa
661 ctcctttact ttgaacacca gggaaacttc aaggagaatt tcctccgcat gctttcagca
721 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttac atatgggcat
781 atgttgccaa actctaaacc aaatactcat tctgatgttt taaatgattt gccctcccat
841 atgtccttcc gagtgagaga cacaaaaaat tccaacacac tattgcaatg aaaataaatt
901 tcctttatta gccagaagtc agatgctcaa ggggcttcat gatgtcccca taatttttgg
961 cagagggaaa aagatcgatc tcagtggtat ttgtgagcca gggcattggc cacaccagcc
1021 accaccttct gataggcagc ctgcacctga ggttaatcac ttagtgatgg tgatggtgat
1081 ggtggtgctc gagatcctcg gggtcttccg gggcgagttc tggctggctt gcacgtgtat
1141 acagctgtgc ggccgcaagc ttttacaaca gcaggcattt tctcttcctc ttcttcacgg
1201 gaggcgggca gaggactgct cggatcgctt cgtcaaacac tgtcttgagg cctcgctgtg
1261 tgagcgccga gcactccagg tattttacag caccaatctc cttagccatg gctagaccct
1321 gcggataggt gatgggagtc agcttcttct ccttcagttt ctcgatcgtg tctttatcat
1381 ccctaagatc aagtttagtt cccactagga tgatgggagt gttgggacag tggtgccgca
1441 cctcaggata ccactttgca cggacgtggt ggaatgatgc aggactcaca agggaaaagc
1501 aaattaagaa cacatctgtt tgcggatagg atagggggcg taatctgtca taatcttcta
1561 gtccagctgt atcccataag cccagattca ccggttttcc atctaccata acattggcag
1621 aataattgtc aaagacagta gggatatatt ctccaggaaa tgcattggtt gtgtaactga
1681 tcagtaggca agttttacct acagctccgt ctcccaccac cacacacttg ataagttctt
1741 ttgccgcctc atcaatattt tctgcagttt tcttaatcag catgactccc tctctctcgg
1801 cagcatctcg gacatgctca gttccatcca actgaacccc aataaagtac tggacatctc
1861 ccttctgatc tcgcataggc tgcaagtgaa agaggttcca gaacttttta ccactctttg
1921 tataattaat cagctgaaca gtgacctctg tttggttatc tatggcatct ctaatttttc
1981 tcactgtcgc gcgatcagtt tcaggacctt gtagaaacct gcagtttctt cccaaaattt
2041 cttcacggct atattctgtc aactgcaaga aactatcgga cgcgaatata atgggattat
2101 ctggcaatct tgggtcagta atgacaaagt tcttctcaat acgttcaagt gtagtagcca
2161 aggatccctt gtacagctcg tccatgccgc cggtggagtg gcggccctcg gcgcgttcgt
2221 actgttccac gatggtgtag tcctcgttgt gggaggtgat gtccaacttg atgttgacgt
2281 tgtaggcgcc gggcagctgc acgggcttct tggccttgta ggtggtcttg acctcagcgt
2341 cgtagtggcc gccgtccttc agcttcagcc tctgcttgat ctcgcccttc agggcgccgt
2401 cctcggggta catccgctcg gaggaggcct cccagcccat ggtcttcttc tgcattacgg
2461 ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt gtagatgaac tcgccgtcct
2521 gcagggagga gtcctgggtc acggtcacca cgccgccgtc ctcgaagttc atcacgcgct
2581 cccacttgaa gccctcgggg aaggacagct tcaagtagtc ggggatgtcg gcggggtgct
2641 tcacgtaggc cttggagccg tacatgaact gaggggacag gatgtcccag gcgaagggca
2701 gggggccacc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct
2761 cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga gccctccatg tgcaccttga
2821 agcgcatgaa ctccttgatg atggccatgt tatcctcctc gcccttgctc acagatccgt
2881 gatggtggtg atggtgtgcc atggtatatc tcctttgatt gtaaataaaa tgtaatttac
2941 agtatagtat tttaattaat atacaaatga tttgataata attcttattt aactataata
3001 tattgtgttg ggttgaatta aaggtcccgg attaactccg gggaattgtt atccgctcac
3061 aattccccta tagtgagtcg tattaatttc ggtccgatcc aattctttgc caaaatgatg
3121 agacagcaca ataaccagca cgttgcccag gagctgtagg aaaaagaaga aggcatgaac
3181 atggttagca gaggctctag agccgccggt cacacgccag aagccgaacc ccgccctgcc
3241 ccgtcccccc cgaaggcagc cgtccccccg cggacagccc cgagccgctc cccccgcaca
3301 aagggcccga tacttacctg cccagtgcct cacgaccaac ttctgcacgt taagttcgag
3361 actggcgtta gcaatttaac tgtgataaac taccgcaata aagcatctag tgatctgacg
3421 gttcactaaa cgacgtctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg
3481 cgtcaacggg gcggggttat tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa
3541 caaactccca ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta
3601 tccacgccca ttggtgtact gccaaaaccg catcagcatg gtaatagcga tgactaatac
3661 gtagatgtac tgccaagtag gaaagtcccg taaggtcatg tactgggcat aatgccaggc
3721 gggccattta ccgtcattga cgtcaatagg gggcggactt ggcatatgat acacttgatg
3781 tactgccaag tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc
3841 tattggcgtt actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg
3901 cggtcagcca ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct
3961 atgaactaat gaccccgtaa ttgattacta ttaataggcc ggccatcagc aactatatat
4021 tgatagacat ttccagtttg tgatattagt ttgtgcgtct cattacaatg gctgttattt
4081 ttaacaacaa acaactgctc gcagacaata gtatagaaaa gggaggtgaa ctgtttttgt
4141 ttaacggttc gtacaacatt ttggaaagtt atgttaatcc ggtgctgcta aaaaatggtg
4201 taattgaact agaagaagct gcgtactatg ccggcaacat attgtacaaa accgacgatc
4261 ccaaattcat tgattatata aatttaataa ttaaagcaac acactccgaa gaactaccag
4321 aaaatagcac tgttgtaaat tacagaaaaa ctatgcgcag cggtactata caccccatta
4381 aaaaagacat atatatttat gacaacaaaa aatttactct atacgataga tacatatatg
4441 gatacgataa taactatgtt aatttttatg aagagaaaaa tgaaaaagag aaggaatacg
4501 aagaagaaga cgacaaggcg tctagtttat gtgaaaataa aattatattg tcgcaaatta
4561 actgtgaatc atttgaaaat gattttaaat attacctcag cgattataac tacgcgtttt
4621 caattataga taatactaca aatgttcttg ttgcgtttgg tttgtatcgt taataaaaaa
4681 caaatttgac atttataatt gttttattat tcaataatta caaataggat tgagaccctt
4741 gcagttgcca gcaaacggac agagcttgtc gacagttttc attttttggc agtttggtgg
4801 tttatacaca tcttgggatt tgattaaaag atgcatgact tcacccacac acttggagtc
4861 gtggttgtag aaatgcgtca gagccttgat taaacatcgg ttttcacact tggcacacga
4921 aaccacgttt gcaatgtaac agtttagtat aaacctcttc ctgtacatac cggcgcgttt
4981 atacacatct tgggatttga ttaaaagatg caggaaacgc gcggacatga gacaataacc
5041 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
5101 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
5161 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
5221 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
5281 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
5341 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
5401 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
5461 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
5521 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
5581 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
5641 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
5701 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
5761 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
5821 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
5881 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaaca
5941 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
6001 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
6061 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
6121 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
6181 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
6241 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
6301 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
6361 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
6421 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
6481 acctctgact tgagcgtcga tttttgtgat gctcgtcagg gggcggagcc tatggaaaaa
6541 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
6601 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
6661 taccgctcgc cgcagccgaa cgaccgagcg cagcgttccg agcttataac ttcgtataat
6721 gtatgctata cgaagttatg cgatcgc
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：EGFP-p65
- 下一篇：pCI-neo.mCherry-NEDD4
