质粒名称: | pcDNA3.1/Puro-CAG-JEDI-2P |
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出品公司: | addgene |
目录编号: | 179458 |
存储实验室: | Francois St-Pierre |
载体骨架基本信息 |
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载体名称: | pcDNA3.1/Puro-CAG |
空载体大小: | 6095 bp |
完整质粒大小: | 7383 bp |
载体类型: | 哺乳细胞表达 |
筛选标记: | 新霉素(G418) |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | JEDI-2P / JEDI2P |
物种来源: | Synthetic |
插入大小: | 1287 bp |
启动子: | CAG |
克隆方法: | 限制性内切酶 |
5’克隆位点: | NheI(未破坏) |
3’克隆位点: | HindIII (未破坏) |
5’测序引物 | AAGGTGGTGGCTGGTGTGGC |
3’测序引物 | TAGAAGGCACAGTCGAGG |
文献引用方法 |
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材料和方法部分: | pcDNA3.1/Puro-CAG-JEDI-2P was a gift from Francois St-Pierre (Addgene plasmid # 179458 ; http://n2t.net/addgene:179458 ; RRID:Addgene_179458) |
参考文献部分: | Sustained deep-tissue voltage recording using a fast indicator evolved for two-photon microscopy. Liu Z, Lu X, Villette V, Gou Y, Colbert KL, Lai S, Guan S, Land MA, Lee J, Assefa T, Zollinger DR, Korympidou MM, Vlasits AL, Pang MM, Su S, Cai C, Froudarakis E, Zhou N, Patel SS, Smith CL, Ayon A, Bizouard P, Bradley J, Franke K, Clandinin TR, Giovannucci A, Tolias AS, Reimer J, Dieudonne S, St-Pierre F. Cell. 2022 Aug 16. pii: S0092-8674(22)00916-3. doi: 10.1016/j.cell.2022.07.013. 10.1016/j.cell.2022.07.013 PubMed 35985322 |
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 pcDNA3.1/Puro-CAG-JEDI-2P 7383 bp ds-DNA circular SYN 03-FEB-2022 DEFINITION Genetically encoded voltage indicator (GEVI) JEDI-2P expressed under strong mammalian promoter (CAG). ACCESSION . VERSION . KEYWORDS pcDNA3.1/Puro-CAG-JEDI-2P SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7383) TITLE Deep, rapid, and prolonged voltage imaging with an indicator optimized for two-photon microscopy JOURNAL Unpublished REFERENCE 2 (bases 1 to 7383) AUTHORS . TITLE Direct Submission JOURNAL Exported Feb 3, 2022 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..7383 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 440..459 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(678..782) /gene="bla" /label=AmpR promoter enhancer 814..1193 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 1472..2488 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 2412..2434 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 2496..2515 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 2648..2657 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(4052..4069) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4058..4282 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4328..4756 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4415..4434) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4625..4646 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4765..4785) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4770..5099 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4950..5085 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5012..5031 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5110..5709 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(5110..5129) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5606..5626 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 5869..5990 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5906..5925) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5960..5979 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6039..6055) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6039..6055) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6052..6074) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6063..6079 /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(6087..6117) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6132..6153 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6270..6287) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6441..7029) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6521..6540) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7200..677) /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" ORIGIN 1 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct 61 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt 121 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc 181 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt 241 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg 301 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg 361 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct 421 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc 481 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt 541 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt 601 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg 661 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat 721 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg 781 cgcacatttc cccgaaaagt gccacctgac gtcgacattg attattgact agttattaat 841 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac 901 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa 961 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt 1021 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc 1081 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat 1141 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg 1201 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt 1261 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg gggggsgcgc 1321 gccrggsggg gsggggsggg gsggggggsg gggsggggsg rggsggrgag gkgcggcggc 1381 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg 1441 gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc ttcgccccgt 1501 gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc 1561 acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct tggtttaatg 1621 acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg agggcccttt 1681 gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga gcgccgcgtg 1741 cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc tttgtgcgct 1801 ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg gggggggctg 1861 cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag ggggtgtggg 1921 cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg agcacggccc 1981 ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc cgggcggggg 2041 gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg agggctcggg 2101 ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga gccgcagcca 2161 ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc aaatctgtgc 2221 ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg cgaagcggtg 2281 cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc gccgccgtcc 2341 ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc gggggggacg 2401 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca 2461 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc 2521 tcatcatttt ggcaaagaat tcctcgagga attcactcct caggtgcagg ctgcctatca 2581 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctgcatga 2641 gctagccgcc accatggaga cgactgtgag gtatgaacag gggtcagagc tcactaaaac 2701 ttcgagctct ccaacagcag atgagcccac gataaagatt gatgatggtc gtgatgaggg 2761 taatgaacaa gacagctgtt ccaataccat taggagaaaa atttccccgt ttgtgatgtc 2821 atttggattc agagtatttg gagttgtgct tatcattgta gacatcatag tggtgattgt 2881 ggatctggcc atcagtgaga agaaaagagg cattagagag attcttgaag gtgtttccct 2941 ggctatagca ctcttcttcc ttgttgatgt tctcatgaga gtgtttgttg aaggcttcaa 3001 gaactatttc cggtccaaac tggttacttt ggatgcagtc atagtagtgg gcactctgct 3061 aattaatatg acctactcct tctctgacct tgctgccttt aacagcgaga acgtgtatat 3121 taccgcggat aaacagaaaa acggcattaa agcgaacttt accgtgcgcc ataacgtgga 3181 agatggcagc gtgcagctgg cggatcatta tcagcagaac accccgattg gcgatggccc 3241 ggtgctgctg ccggataacc attatctgag ccaccagacc gtgctgagca aagatccgaa 3301 cgaaaaacgc gatcacatgg tgctgctgga atttgtgacc gcagcgggca ttacacacgg 3361 catggatgaa ctgtatggcg gcaccggcgg cagcgcgagc cagggcgaag aactgtttac 3421 cggcgtggtg ccgattctgg tggaactgga tggcgatgtg aacggccata aatttagcgt 3481 gcgcggcgaa ggcgaaggcg atgcgaccat tggcaaactg accctgaaat ttatttgcac 3541 caccggcaaa ctaccggtgc cgtggccgac cctggtgacc accttaacct atggcgtgca 3601 gtgctttagc cgctatccgg atcatatgaa acgccatgat ttttttaaaa gcgcgatgcc 3661 ggaaggctat gtgcaggaac gcaccattag ctttaaagat gatggcaaat ataaaacccg 3721 cgcggtggtg aaatttgaag gcgataccct ggtgaaccgc attgaactga aaggcaccga 3781 ttttaaagaa gatggcaaca ttctggggca taaactggaa tatgatacag atcagatgcc 3841 gcacatggtt actcttcttc gagttctgaa aattgttatc ttaataagaa tatttcgcct 3901 ggcttcacag aagaaacaac ttgaagtggt aacctaatga aagcttggta ccgagctcgg 3961 atccactagt ccagtgtggt ggaattctgc agatatccag cacagtggcg gccgctcgag 4021 tctagagggc ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca 4081 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc 4141 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg 4201 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct 4261 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg 4321 tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc 4381 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt 4441 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc 4501 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt 4561 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt 4621 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt 4681 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa 4741 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag 4801 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg 4861 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 4921 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc 4981 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg 5041 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa 5101 agctcccgca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc 5161 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc 5221 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc 5281 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc 5341 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag 5401 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg 5461 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag 5521 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc 5581 gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct cggcttcacc 5641 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc 5701 ggtgcctgat tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat 5761 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg 5821 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt 5881 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 5941 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt 6001 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga 6061 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc 6121 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc 6181 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc 6241 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 6301 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 6361 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 6421 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 6481 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 6541 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 6601 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 6661 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 6721 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 6781 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 6841 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 6901 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 6961 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 7021 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 7081 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 7141 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 7201 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 7261 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 7321 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 7381 cag //