| 质粒名称: | GLP1R-Tango |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 66295 |
| 存储实验室: | Bryan Roth |
|
载体骨架基本信息 |
|
| 载体名称: | Tango |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 6632 bp |
| 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: | GLP1R |
| 物种来源: | -- |
| 插入大小: | 1389 bp |
| 基因突变: | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | GLP1R-Tango was a gift from Bryan Roth (Addgene plasmid # 66295 ; http://n2t.net/addgene:66295 ; RRID:Addgene_66295) |
| 参考文献部分: | PRESTO-Tango as an open-source resource for interrogation of the druggable human GPCRome. Kroeze WK, Sassano MF, Huang XP, Lansu K, McCorvy JD, Giguere PM, Sciaky N, Roth BL. Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014. Epub 2015 Apr 20. 10.1038/nsmb.3014 PubMed 25895059 |
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 GLP1R-Tango 8027 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expression of G protein-coupled receptors for PRESTO-Tango: parallel
receptorome expression and screening via transcriptional output,
with transcriptional activation following arrestin translocation.
ACCESSION .
VERSION .
KEYWORDS GLP1R-Tango
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8027)
AUTHORS Kroeze WK, Sassano MF, Huang XP, Lansu K, McCorvy JD, Giguere PM,
Sciaky N, Roth BL
TITLE PRESTO-Tango as an open-source resource for interrogation of the
druggable human GPCRome.
JOURNAL Nat Struct Mol Biol. 2015 May;22(5):362-9. doi: 10.1038/nsmb.3014.
Epub 2015 Apr 20.
PUBMED 25895059
REFERENCE 2 (bases 1 to 8027)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8027
/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 2582..3583
/codon_start=1
/product="tetracycline-controlled transactivator,
comprising a fusion of the tetracycline repressor TetR with
the C-terminal activation domain of herpes simplex virus
VP16"
/label=tTA
/note="In the Tet-Off(R) system, tTA binds to the
Tet-responsive element and stimulates transcription only in
the absence of tetracycline or doxycycline."
/translation="RLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVKN
KRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRPT
EKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTDS
MPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIEG
LLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHADA
LDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
primer_bind complement(2649..2668)
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
primer_bind complement(3621..3638)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3627..3851
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3897..4325
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3984..4003)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4194..4215
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4334..4354)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4339..4668
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4519..4654
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4581..4600
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4735..5529
/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(4789..4808)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5399..5418
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5703..5824
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5740..5759)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5794..5813
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5873..5889)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5873..5889)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5886..5908)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5897..5913
/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(5921..5951)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5966..5987
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6104..6121)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6275..6860)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6355..6374)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7031..7891)
/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 7654..7673
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7892..7996)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 gctggctagc
901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
961 agatatccag cacagtggcg gccgcgccac catgaagacg atcatcgccc tgagctacat
1021 cttctgcctg gtattcgccg actacaagga cgatgatgac gccagcatcg atatggcagg
1081 tgcccctgga cccctcaggc tcgcactgct gcttctgggg atggtcggac gagccggacc
1141 cagacctcag ggcgctaccg tgagtctttg ggagaccgtc cagaagtggc gggagtatcg
1201 ccgccagtgt caacgcagcc tgacagaaga tcctcctcca gccaccgact tgttttgcaa
1261 taggacattt gatgagtatg cctgctggcc agacggcgag cccggctcat tcgtgaatgt
1321 gtcatgtcca tggtacctgc catgggcatc atctgtgcct caaggacacg tctacaggtt
1381 ctgcacagct gagggcctct ggctgcaaaa agataacagt tcccttccat ggcgcgatct
1441 tagcgagtgc gaggaatcta agaggggcga gcgcagttcc cccgaggaac agctgctgtt
1501 tctgtacatt atctacaccg tcggctacgc gctgtccttt tccgctctgg ttatcgctag
1561 tgccatcctt ttggggtttc gccaccttca ctgcacaaga aattacatcc acttgaatct
1621 ctttgcaagt tttattctgc gggccttgag tgtgttcatt aaggacgctg ccctgaagtg
1681 gatgtactct accgcggccc aacagcacca atgggatgga ctcctcagct atcaggattc
1741 tctttcctgt cgactcgtgt ttctgcttat gcagtactgt gtggcagcca attattattg
1801 gctgctggtc gaaggagtat atctttacac actgctcgca ttctccgtgc tgagtgaaca
1861 gtggattttc aggctttacg tgtccattgg atggggtgtg cctctgctgt tcgtggtgcc
1921 ttggggcatc gtcaagtacc tgtatgaaga tgaaggctgt tggaccagaa actctaacat
1981 gaactactgg ctcatcatac gcctgcccat actcttcgcg attggagtca actttcttat
2041 atttgtgaga gtgatctgta tcgtcgtgtc aaagctgaaa gctaacctta tgtgcaaaac
2101 cgacattaag tgcagactgg ctaagtccac tctcaccctc atcccgctgc tcggaacaca
2161 tgaagtgata tttgccttcg tgatggacga gcatgcgaga ggcacactgc ggtttatcaa
2221 actgttcaca gagctgtctt tcactagttt ccaaggattg atggtggcta ttctctactg
2281 cttcgtgaac aatgaggtcc aacttgaatt tcggaagtca tgggaacggt ggcggcttga
2341 acacctccat attcagcgcg atagttctat gaagccactt aaatgtccca cttccagcct
2401 gagttccgga gccaccgccg gctcttcaat gtatacagcc acttgtcagg cgtcctgctc
2461 tatcgatacc ggtggacgca ccccacccag cctgggtccc caagatgagt cctgcaccac
2521 cgccagctcc tccctggcca aggacacttc atcgaccggt gagaacctgt acttccagct
2581 aagattagat aaaagtaaag tgattaacag cgcattagag ctgcttaatg aggtcggaat
2641 cgaaggttta acaacccgta aactcgccca gaagctaggt gtagagcagc ctacattgta
2701 ttggcatgta aaaaataagc gggctttgct cgacgcctta gccattgaga tgttagatag
2761 gcaccatact cacttttgcc ctttagaagg ggaaagctgg caagattttt tacgtaataa
2821 cgctaaaagt tttagatgtg ctttactaag tcatcgcgat ggagcaaaag tacatttagg
2881 tacacggcct acagaaaaac agtatgaaac tctcgaaaat caattagcct ttttatgcca
2941 acaaggtttt tcactagaga atgcattata tgcactcagc gctgtggggc attttacttt
3001 aggttgcgta ttggaagatc aagagcatca agtcgctaaa gaagaaaggg aaacacctac
3061 tactgatagt atgccgccat tattacgaca agctatcgaa ttatttgatc accaaggtgc
3121 agagccagcc ttcttattcg gccttgaatt gatcatatgc ggattagaaa aacaacttaa
3181 atgtgaaagt gggtccgcgt acagccgcgc gcgtacgaaa aacaattacg ggtctaccat
3241 cgagggcctg ctcgatctcc cggacgacga cgcccccgaa gaggcggggc tggcggctcc
3301 gcgcctgtcc tttctccccg cgggacacac gcgcagactg tcgacggccc ccccgaccga
3361 tgtcagcctg ggggacgagc tccacttaga cggcgaggac gtggcgatgg cgcatgccga
3421 cgcgctagac gatttcgatc tggacatgtt gggggacggg gattccccgg gtccgggatt
3481 taccccccac gactccgccc cctacggcgc tctggatatg gccgacttcg agtttgagca
3541 gatgtttacc gatgcccttg gaattgacga gtacggtggg tagctcgagt ctagagggcc
3601 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
3661 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
3721 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
3781 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
3841 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
3901 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
3961 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
4021 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
4081 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
4141 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
4201 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
4261 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
4321 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
4381 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
4441 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
4501 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
4561 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
4621 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
4681 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
4741 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
4801 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
4861 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
4921 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
4981 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
5041 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
5101 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
5161 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
5221 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
5281 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
5341 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
5401 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
5461 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
5521 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
5581 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
5641 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
5701 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
5761 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
5821 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
5881 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
5941 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
6001 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
6061 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
6121 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
6181 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
6241 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
6301 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
6361 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
6421 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
6481 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
6541 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
6601 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
6661 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
6721 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
6781 gatccggcaa acaaaccacc gctggtagcg gtttttttgt ttgcaagcag cagattacgc
6841 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
6901 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
6961 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
7021 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
7081 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
7141 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
7201 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
7261 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
7321 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
7381 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
7441 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
7501 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
7561 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
7621 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
7681 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
7741 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
7801 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
7861 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
7921 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
7981 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
