| 质粒名称: | Clover-mRuby2-FRET-10 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 58169 |
| 存储实验室: | Michael Davidson |
|
载体骨架基本信息 |
|
| 载体名称: | -- |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: |
Clover（插入C端子）
mRuby2（插入件上的N端子）
|
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | Clover-mRuby2-FRET-10 was a gift from Michael Davidson (Addgene plasmid # 58169 ; http://n2t.net/addgene:58169 ; RRID:Addgene_58169) |
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 Clover-mRuby2-FRET-10 5395 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Localization: Cytosolic FRET, Excitation: , Emission: .
ACCESSION .
VERSION .
KEYWORDS Clover-mRuby2-FRET-10
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5395)
TITLE FRET Indicator - Live and Fixed Cells
REFERENCE 2 (bases 1 to 5395)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5395
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 305..508
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 459..479
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 535..544
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 541..1251
/codon_start=1
/product="monomeric red fluorescent protein derived from
mRuby, with improved photophysical properties (Lam et al.,
2012)"
/label=mRuby2
/translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
AKFAGLGGGMDELYK"
regulatory 1276..1285
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1282..2001
/codon_start=1
/product="monomeric derivative of the bright green-yellow
fluorescent protein Clover"
/label=mClover2
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1327..1348)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind 1935..1956
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 2123..2244
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2160..2179)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2214..2233
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2251..2706)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2388..2409)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2600..2619
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2733..2837
/gene="bla"
/label=AmpR promoter
promoter 2839..3196
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2862..2882)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3047..3182
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3109..3128
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3231..4025
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3285..3304)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3895..3914
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4213..4232)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4257..4304
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4633..5221
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5122..5141
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccacc
541 atggtgtcta agggcgaaga gctgatcaag gaaaatatgc gtatgaaggt ggtcatggaa
601 ggttcggtca acggccacca attcaaatgc acaggtgaag gagaaggcaa tccgtacatg
661 ggaactcaaa ccatgaggat caaagtcatc gagggaggac ccctgccatt tgcctttgac
721 attcttgcca cgtcgttcat gtatggcagc cgtactttta tcaagtaccc gaaaggcatt
781 cctgatttct ttaaacagtc ctttcctgag ggttttactt gggaaagagt tacgagatac
841 gaagatggtg gagtcgtcac cgtcatgcag gacaccagcc ttgaggatgg ctgtctcgtt
901 taccacgtcc aagtcagagg ggtaaacttt ccctccaatg gtcccgtgat gcagaagaag
961 accaagggtt gggagcctaa tacagagatg atgtatccag cagatggtgg tctgagggga
1021 tacactcata tggcactgaa agttgatggt ggtggccatc tgtcttgctc tttcgtaaca
1081 acttacaggt caaaaaagac cgtcgggaac atcaagatgc ccggtatcca tgccgttgat
1141 caccgcctgg aaaggttaga ggaaagtgac aatgaaatgt tcgtagtaca acgcgaacac
1201 gcagttgcca agttcgccgg gcttggtggt gggatggacg agctgtacaa gtccggactc
1261 agatccccac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
1321 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt ccgcggcgag
1381 ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag
1441 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcgtggc ctgcttcagc
1501 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
1561 gtccaggagc gcaccatctc tttcaaggac gacggtacct acaagacccg cgccgaggtg
1621 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
1681 gacggcaaca tcctggggca caagctggag tacaacttca acagccacaa cgtctatatc
1741 acggccgaca agcagaagaa cggcatcaag gctaacttca agatccgcca caacgttgag
1801 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
1861 gtgctgctgc ccgacaacca ctacctgagc catcagtccg ccctgagcaa agaccccaac
1921 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
1981 atggacgagc tgtacaagta agcggccgcg actctagatc ataatcagcc ataccacatt
2041 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
2101 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
2161 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
2221 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
2281 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
2341 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
2401 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
2461 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
2521 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
2581 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
2641 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
2701 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
2761 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
2821 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
2881 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
2941 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
3001 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
3061 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
3121 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
3181 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
3241 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
3301 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
3361 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
3421 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
3481 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
3541 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
3601 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
3661 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
3721 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
3781 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
3841 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
3901 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
3961 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
4021 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
4081 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
4141 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
4201 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
4261 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
4321 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
4381 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
4441 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
4501 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
4561 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
4621 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
4681 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
4741 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
4801 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
4861 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
4921 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
4981 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
5041 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
5101 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
5161 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
5221 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
5281 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt
5341 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccg
//
