| 质粒名称: | FRB-NLuc |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 31181 |
| 存储实验室: | David Piwnica-Worms |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1-V5/HIS TOPO |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5523 bp |
| 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: | Rapamycin-binding domain and the N-terminus of Firefly Luciferase,FRB,FLUC |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1608 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | TOPO克隆 |
| 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: | FRB-NLuc was a gift from David Piwnica-Worms (Addgene plasmid # 31181 ; http://n2t.net/addgene:31181 ; RRID:Addgene_31181) |
| 参考文献部分: | Kinetics of regulated protein-protein interactions revealed with firefly luciferase complementation imaging in cells and living animals. Luker KE, Smith MC, Luker GD, Gammon ST, Piwnica-Worms H, Piwnica-Worms D. Proc Natl Acad Sci U S A. 2004 Aug 17. 101(33):12288-93. 10.1073/pnas.0404041101 PubMed 15284440 |
买家导航
FRBNLuc载体基本信息
FRBNLuc载体图谱质粒图谱和多克隆位点信息
FRBNLuc载体简介
FRBNLuc载体序列
LOCUS FRB-NLuc 7145 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS FRB-NLuc
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7145)
AUTHORS Luker KE, Smith MC, Luker GD, Gammon ST, Piwnica-Worms H,
Piwnica-Worms D
TITLE Kinetics of regulated protein-protein interactions revealed with
firefly luciferase complementation imaging in cells and living
animals.
JOURNAL Proc Natl Acad Sci U S A. 2004 Aug 17. 101(33):12288-93.
PUBMED 15284440
REFERENCE 2 (bases 1 to 7145)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7145
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 965..1234
/codon_start=1
/product="FKBP-rapamycin binding domain of human FRAP"
/label=FRB
/translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
primer_bind complement(1343..1361)
/label=LucNrev
/note="Firefly luciferase, reverse primer"
CDS 2642..2683
/codon_start=1
/product="epitope tag from simian virus 5"
/label=V5 tag
/translation="GKPIPNPLLGLDST"
CDS 2693..2710
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(2733..2750)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2739..2963
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3009..3437
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3096..3115)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3306..3327
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3446..3466)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3451..3781
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3632..3767
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3694..3713
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3848..4642
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3902..3921)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4512..4531
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4818..4939
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4855..4874)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4909..4928
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4988..5004)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4988..5004)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5001..5023)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5012..5028
/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 complement(5036..5066)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5081..5102
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5219..5236)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5390..5978)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5470..5489)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6149..7009)
/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 6772..6791
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7010..7114)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
901 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattgcc cttacaccac
961 catggagatg tggcatgaag gcctggaaga ggcatctcgt ttgtactttg gggaaaggaa
1021 cgtgaaaggc atgtttgagg tgctggagcc cttgcatgct atgatggaac ggggccccca
1081 gactctgaag gaaacatcct ttaatcaggc ctatggtcga gatttaatgg aggcccaaga
1141 gtggtgcagg aagtacatga aatcagggaa tgtcaaggac ctcacccaag cctgggacct
1201 ctattatcat gtgttccgac gaatctcaaa gcagcagatc tcgtacgcgt cccggggcgg
1261 tggctcatct ggcggaggtg aagacgccaa aaacataaag aaaggcccgg cgccattcta
1321 tccgctggaa gatggaaccg ctggagagca actgcataag gctatgaaga gatacgccct
1381 ggttcctgga acaattgctt ttacagatgc acatatcgag gtggacatca cttacgctga
1441 gtacttcgaa atgtccgttc ggttggcaga agctatgaaa cgatatgggc tgaatacaaa
1501 tcacagaatc gtcgtatgca gtgaaaactc tcttcaattc tttatgccgg tgttgggcgc
1561 gttatttatc ggagttgcag ttgcgcccgc gaacgacatt tataatgaac gtgaattgct
1621 caacagtatg ggcatttcgc agcctaccgt ggtgttcgtt tccaaaaagg ggttgcaaaa
1681 aattttgaac gtgcaaaaaa agctcccaat catccaaaaa attattatca tggattctaa
1741 aacggattac cagggatttc agtcgatgta cacgttcgtc acatctcatc tacctcccgg
1801 ttttaatgaa tacgattttg tgccagagtc cttcgatagg gacaagacaa ttgcactgat
1861 catgaactcc tctggatcta ctggtctgcc taaaggtgtc gctctgcctc atagaactgc
1921 ctgcgtgaga ttctcgcatg ccagagatcc tatttttggc aatcaaatca ttccggatac
1981 tgcgatttta agtgttgttc cattccatca cggttttgga atgtttacta cactcggata
2041 tttgatatgt ggatttcgag tcgtcttaat gtatagattt gaagaagagc tgtttctgag
2101 gagccttcag gattacaaga ttcaaagtgc gctgctggtg ccaaccctat tctccttctt
2161 cgccaaaagc actctgattg acaaatacga tttatctaat ttacacgaaa ttgcttctgg
2221 tggcgctccc ctctctaagg aagtcgggga agcggttgcc aagaggttcc atctgccagg
2281 tatcaggcaa ggatatgggc tcactgagac tacatcagct attctgatta cacccgaggg
2341 ggatgataaa ccgggcgcgg tcggtaaagt tgttccattt tttgaagcga aggttgtgga
2401 tctggatacc gggaaaacgc tgggcgttaa tcaaagaggc gaactgtgtg tgagaggtcc
2461 tatgattatg tccggttatg taaacaatcc ggaagcgacc aacgccttga ttgacaagga
2521 tggatgataa cattctggta gctaggtaat gcataactag ctaaggtacc caattaaggg
2581 caattctgca gatatccagc acagtggcgg ccgctcgagt ctagagggcc cgcggttcga
2641 aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtcatcatca
2701 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
2761 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
2821 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
2881 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
2941 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
3001 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
3061 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
3121 tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc ctttagggtt
3181 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
3241 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
3301 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
3361 tgatttataa gggattttgg ggatttcggc ctattggtta aaaaatgagc tgatttaaca
3421 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
3481 ggctccccag gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
3541 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
3601 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
3661 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
3721 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
3781 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
3841 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
3901 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
3961 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
4021 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
4081 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
4141 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
4201 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
4261 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
4321 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
4381 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
4441 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
4501 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
4561 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
4621 ccttcttgac gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga
4681 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
4741 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
4801 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
4861 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
4921 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
4981 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
5041 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
5101 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
5161 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
5221 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
5281 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
5341 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
5401 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
5461 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
5521 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
5581 ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
5641 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
5701 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
5761 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
5821 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
5881 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
5941 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
6001 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
6061 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
6121 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
6181 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
6241 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
6301 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
6361 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
6421 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
6481 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
6541 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
6601 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
6661 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
6721 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
6781 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
6841 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
6901 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
6961 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
7021 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
7081 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
7141 acgtc
//
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