| 质粒名称: | pAAV-UBC-S2-cAMPr |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 160723 |
| 存储实验室: | Edward Boyden |
|
载体骨架基本信息 |
|
| 载体名称: | pAAV-UBC |
| 载体出品公司: | Epoch Life Science |
| 空载体大小: | 4178 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | S2-cAMPr,S2-cAMPr-HA,SiRI-S2-cAMPr |
| 物种来源: | Synthetic |
| 插入大小: | 3558 bp |
| 基因突变: | -- |
| GenBank编号 | MT449929 |
| 启动子: | 人泛素C（UBC）启动子 |
| 克隆方法: | Gibson克隆 |
| 5’测序引物 | GATCGCTGTGATCGTCACTTGG |
| 3’测序引物 | WPRE-R |
|
文献引用方法 |
|
| 材料和方法部分: | pAAV-UBC-S2-cAMPr was a gift from Edward Boyden (Addgene plasmid # 160723 ; http://n2t.net/addgene:160723 ; RRID:Addgene_160723) |
| 参考文献部分: | Spatial Multiplexing of Fluorescent Reporters for Imaging Signaling Network Dynamics. Linghu C, Johnson SL, Valdes PA, Shemesh OA, Park WM, Park D, Piatkevich KD, Wassie AT, Liu Y, An B, Barnes SA, Celiker OT, Yao CC, Yu CJ, Wang R, Adamala KP, Bear MF, Keating AE, Boyden ES. Cell. 2020 Nov 17. pii: S0092-8674(20)31399-4. doi: 10.1016/j.cell.2020.10.035. 10.1016/j.cell.2020.10.035 PubMed 33232692 |
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 pAAV-UBC-S2-cAMPr 7742 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Signaling reporter island (SiRI) construct for spatially multiplexed
imaging. Contains GFP-based fluorescent reporter cAMPr (cAMP
indicator), HA tag, and S2 protein scaffold..
ACCESSION .
VERSION .
KEYWORDS pAAV-UBC-S2-cAMPr
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7742)
AUTHORS Linghu C, Johnson SL, Valdes PA, Shemesh OA, Park WM, Park D,
Piatkevich KD, Wassie AT, Liu Y, An B, Barnes SA, Celiker OT, Yao
CC, Yu CJ, Wang R, Adamala KP, Bear MF, Keating AE, Boyden ES
TITLE Spatial Multiplexing of Fluorescent Reporters for Imaging Signaling
Network Dynamics.
JOURNAL Cell. 2020 Nov 17. pii: S0092-8674(20)31399-4. doi:
10.1016/j.cell.2020.10.035.
PUBMED 33232692
REFERENCE 2 (bases 1 to 7742)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7742
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..399
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 2621..2642
/label=EGFP-C
/note="EGFP, forward primer"
CDS 3633..3659
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
misc_feature 3984..4572
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4037..4057)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4455..4466)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
polyA_signal 4615..4736
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4626..4645)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 4680..4699
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
repeat_region 4898..5027
/label=AAV2 ITR
primer_bind complement(5093..5110)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5264..5852)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5344..5363)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6023..6883)
/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 6646..6665
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6884..6988)
/gene="bla"
/label=AmpR promoter
rep_origin complement(7014..7469)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7151..7172)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 7363..7382
/label=F1ori-R
/note="F1 origin, reverse primer"
repeat_region 7541..7670
/label=AAV2 ITR
ORIGIN
1 ggcctccgcg ccgggttttg gcgcctcccg cgggcgcccc cctcctcacg gcgagcgctg
61 ccacgtcaga cgaagggcgc agcgagcgtc ctgatccttc cgcccggacg ctcaggacag
121 cggcccgctg ctcataagac tcggccttag aaccccagta tcagcagaag gacattttag
181 gacgggactt gggtgactct agggcactgg ttttctttcc agagagcgga acaggcgagg
241 aaaagtagtc ccttctcggc gattctgcgg agggatctcc gtggggcggt gaacgccgat
301 gattatataa ggacgcgccg ggtgtggcac agctagttcc gtcgcagccg ggatttgggt
361 cgcggttctt gtttgtggat cgctgtgatc gtcacttggc gactagtgcc accatgaagc
421 ccaccacaat cagcctgctg cagaagtaca agcaggagaa gaagcggttc gccaccatca
481 cagcttacga ctactccttc gccaagctgt ttgctgatga gggactgaac gtgatgctgg
541 tgggagactc tctgggcatg accgtgcagg gacacgacag caccctgcca gtgacagtgg
601 ccgatatcgc ttaccacaca gccgctgtga ggagaggagc ccctaactgc ctgctgctgg
661 ctgatctgcc cttcatggcc tacgctaccc ctgagcaggc ctttgagaac gccgctacag
721 tgatgagggc cggcgctaac atggtgaaga tcgagggcgg agagtggctg gtggagaccg
781 tgcagatgct gacagagaga gccgtgccag tgtgcggaca cctgggactg accccacagt
841 ctgtgaacat ctttggcgga tacaaggtgc agggaagggg actgctggct ggactgcagc
901 tgctgtccga cgctctggct ctggaggctg ctggagctca gctgctggtg ctggagtgcg
961 tgcctgtgga gctggctaag cgcatcacag aggccctggc tatcccagtg atcggaatcg
1021 gagctggaaa cgtgaccgac ggacagatcc tggtcatgca cgatgctttc ggcatcacag
1081 gcggacacat cccaaagttc gccaagaact ttctggctga gaccggcgat atcagggctg
1141 ctgtgcgcca gtacatggct gaggtggagt ccggagtgta cccaggagag gagcacagct
1201 ttcactctag aggaggttct ggtggaaccg gtggaagtgg agggacagga ggttcaggtg
1261 gtactggtgg ctctggagga acaggaggct ccggaggtac aggagggtca ggagggactg
1321 gaggctctgg aggtactggc ggttcaggag gtaccatggg caacgccgcc gccgccaaga
1381 agggcagcga gcaggagagc gtgaaagagt tcctagccaa agccaaggaa gatttcctga
1441 aaaaatggga gaccccttct cagaatacag cccagttgga tcagtttgat agaatcaaga
1501 cccttggcac cggctccttt gggcgagtga tgctggtgaa gcacaaggag agtgggaacc
1561 actacgccat gaagatctta gacaagcaga aggtggtgaa gctaaagcag atcgagcaca
1621 ctctgaatga gaagcgcatc ctgcaggccg tcaacttccc gttcctggtc aaacttgaat
1681 tctccttcaa ggacaactca aacctgtaca tggtcatgga gtatgtagct ggtggcgaga
1741 tgttctccca cctacggcgg attggaaggt tcagcgagcc ccatgcccgt ttctacgcgg
1801 cgcagatcgt cctgaccttt gagtatctgc actccctgga cctcatctac cgggacctga
1861 agcccgagaa tcttctcatc gaccagcagg gctatattca ggtgacagac ttcggttttg
1921 ccaagcgtgt gaaaggccgt acttggacct tgtgtgggac ccctgagtac ttggcccccg
1981 agattatcct gagcaaaggc tacaacaagg ctgtggactg gtgggctctc ggagtcctca
2041 tctacgagat ggctgctggt tacccaccct tcttcgctga ccagcctatc cagatctatg
2101 agaaaatcgt ctctgggaag gtgcggttcc catcccactt cagctctgac ttgaaggacc
2161 tgctgcggaa ccttctgcaa gtggatctca ccaagcgctt tgggaacctc aagaacgggg
2221 tcaatgacat caagaaccac aagtggtttg ccacgactga ctggattgcc atctatcaga
2281 gaaaggtgga agctcccttc ataccaaagt ttaaaggccc tggggacacg agtaactttg
2341 acgactatga ggaggaagag atccgggtct ccatcaatga gaagtgtggc aaggagttta
2401 ctgagtttcc cgggaacgtc tatatcaagg ccgacgagca gaagaacggc atcaaggcgt
2461 acttcaagat ccgccacaac atcgagggcg gcggcgtgca gctcgcctac cactaccagc
2521 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc
2581 agtccatgct ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
2641 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt accggaggga
2701 gcgaatccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atccaggtcg
2761 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggt gagggcgatg
2821 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
2881 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
2941 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacatc caggagcgca
3001 ccatcttctt caagggcgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
3061 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
3121 tggggcacaa gctggagtac aacaccgccc cccccgcatg caagggccgg cggcgacgcg
3181 gggccatcag cgctgaggtc tacaccgagg aggatgccgc gtcctatgtt cggaaggtta
3241 taccgaaaga ttataagaca atggctgctt tagctaaagc cattgaaaag aacgtactgt
3301 tttcacatct tgatgataac gagagaagtg acatttttga cgccatgttc ccggtttcct
3361 ttattgctgg agagactgtt attcagcagg gtgacgaagg ggataacttc tacgtgattg
3421 accaaggaga gatggacgtc tatgtcaaca atgaatgggc aaccagtgtt ggggaaggag
3481 ggagcttcgg ggagcttgct ctgatttacg ggactcctcg agcggccact gtcaaggcca
3541 agacgaacgt gaaactgtgg ggcattgacc gggacagcta cagaaggatc ctcatgggaa
3601 gcacgctgag aaagcggaag atgggatccg gttacccata cgacgttcca gactacgctg
3661 gaggttctgg aggttctggt ggtactggtg gttctggagg atctggtggt acaggaggta
3721 gcggaggttc tggtggaacc ggaagcagag tgtacgagtc tgagaaactg gctagagagg
3781 cagacaagct ggctcagaag agcgaggata tggctagaga ggcagataag caggccagga
3841 gggctgagga aagacctgac agagaagaga ttgctaggct cgctgccatc attgctagga
3901 tggtggctct caactccaga atcgcaatgc tcatggcacg catgatcatg ctgaactctc
3961 aagagagcta aaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
4021 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
4081 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
4141 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
4201 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
4261 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
4321 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
4381 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
4441 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
4501 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
4561 ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat
4621 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
4681 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
4741 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
4801 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac
4861 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
4921 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
4981 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa
5041 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
5101 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
5161 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
5221 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
5281 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
5341 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
5401 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
5461 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
5521 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
5581 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
5641 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
5701 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
5761 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
5821 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
5881 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
5941 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
6001 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
6061 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
6121 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
6181 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
6241 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
6301 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
6361 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
6421 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
6481 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
6541 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
6601 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
6661 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
6721 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
6781 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
6841 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
6901 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
6961 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
7021 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
7081 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
7141 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
7201 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
7261 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
7321 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
7381 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
7441 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
7501 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctgcgcgctc gctcgctcac
7561 tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
7621 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct tgtagttaat
7681 gattaacccg ccatgctact tatctacgta gccatgctct aggaagatcg taggttaatt
7741 aa
//
