质粒名称: | pGP-pcDNA3.1 Puro-CAG-Voltron2 |
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出品公司: | addgene |
目录编号: | 172909 |
存储实验室: | GENIE Project |
载体骨架基本信息 |
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载体名称: | pcDNA3.1 |
完整质粒大小: | 7755 bp |
载体类型: | 哺乳细胞表达 |
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: | Voltron2 / Voltron2 insert /variant 421.2096 |
物种来源: | Synthetic |
插入大小: | 1659 bp |
基因突变: | A122D |
启动子: | CAG |
克隆方法: | Gibson Cloning |
5’测序引物 | GGTTCGGCTTCTGGCGTGTGACC |
3’测序引物 | TTAAAGCAGCGTATCCACAT |
文献引用方法 |
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材料和方法部分: | pGP-pcDNA3.1 Puro-CAG-Voltron2 was a gift from GENIE Project (Addgene plasmid # 172909 ; http://n2t.net/addgene:172909 ; RRID:Addgene_172909) |
参考文献部分: | Sensitivity optimization of a rhodopsin-based fluorescent voltage indicator. Abdelfattah AS, Zheng J, Singh A, Huang YC, Reep D, Tsegaye G, Tsang A, Arthur BJ, Rehorova M, Olson CVL, Shuai Y, Zhang L, Fu TM, Milkie DE, Moya MV, Weber TD, Lemire AL, Baker CA, Falco N, Zheng Q, Grimm JB, Yip MC, Walpita D, Chase M, Campagnola L, Murphy GJ, Wong AM, Forest CR, Mertz J, Economo MN, Turner GC, Koyama M, Lin BJ, Betzig E, Novak O, Lavis LD, Svoboda K, Korff W, Chen TW, Schreiter ER, Hasseman JP, Kolb I. Neuron. 2023 Mar 28:S0896-6273(23)00205-2. doi: 10.1016/j.neuron.2023.03.009. 10.1016/j.neuron.2023 |
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LOCUS pGP-pcDNA3.1 Puro-CAG-Voltron2 7755 bp ds-DNA circular SYN 26-AUG-2021 DEFINITION Mammalian expression of voltage sensor. ACCESSION . VERSION . KEYWORDS pGP-pcDNA3.1 Puro-CAG-Voltron2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7755) AUTHORS GENIE Project TITLE GENIE voltage sensors JOURNAL unpublished REFERENCE 2 (bases 1 to 7755) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 26, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..7755 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 57..76 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 209..218 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" primer_bind complement(1987..2004) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1993..2217 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2263..2691 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2350..2369) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2560..2581 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2700..2720) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2705..3034 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 2885..3020 /label=SV40 ori /note="SV40 origin of replication" primer_bind 2947..2966 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3045..3644 /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(3045..3064) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3541..3561 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 3804..3925 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3841..3860) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3895..3914 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(3974..3990) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3974..3990) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3987..4009) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3998..4014 /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(4022..4052) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4067..4088 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4205..4222) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4376..4964) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4456..4475) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5135..5995) /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 5758..5777 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5996..6100) /gene="bla" /label=AmpR promoter enhancer 6132..6511 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 6788..49 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 7728..7750 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" ORIGIN 1 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa 61 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctcgagg aattcactcc 121 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa 181 ataccactga gatctgcatg agctagccgc caccatggct gacgtggaaa ccgagaccgg 241 catgattgca cagtggattg tctttgctat tatggctgct gctgctattg cttttggagt 301 ggctgtgcac tttcggcctt cagagctgaa gagcgcatac tatatcaaca ttgccatctg 361 cactatcgcc gctaccgctt actatgcaat ggccgtgaac taccaggacc tgacaatgaa 421 tggtgaaagg caggtggtct acgcaagata tattaactgg gtgctgacca caccactgct 481 cctgctcgat ctcatcgtca tgaccaagat gggcggagtg atgatttctt gggtcatcgg 541 cgcagacatt ttcatgatcg tgtttggtat tctgggcgac ttcgaggatg aacacaagtt 601 caaatgggtg tactttatcg ctggatgtgt gatgcaggca gtcctgacat acgggatgta 661 taacgccact tggaaagacg atctgaagaa aagccccgag taccatagct cctatgtcag 721 tctgctcgtc ttcctgtcaa tcctctgggt gttttatcct gtcgtgtggg ctttcgggtc 781 tggtagtggc gtgctgtccg tcgacaatga ggccattctc atgggaatcc tggatgtgct 841 cgctaagcca ctgtttggaa tggggtgcct cattgcccat gagactatct tcaagatcgg 901 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt 961 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc 1021 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga 1081 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt 1141 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca 1201 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg 1261 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc 1321 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca 1381 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga 1441 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt 1501 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata 1561 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt 1621 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt 1681 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga 1741 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccacta agagcaggat 1801 caccagcgag ggcgagtaca tccccctgga ccagatcgac atcaacgtgt tctgctacga 1861 gaacgaggtg taatgaaagc ttggtaccga gctcggatcc actatccagt gtggtggaat 1921 tctgcagata tccagcacag tggcggccgc tcgagtctag agggcccgtt taaacccgct 1981 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc 2041 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg 2101 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca 2161 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt 2221 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg 2281 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc 2341 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc 2401 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg 2461 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg 2521 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg 2581 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt 2641 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg 2701 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca 2761 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg 2821 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc 2881 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat 2941 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg 3001 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgcatgacc gagtacaagc 3061 ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg 3121 cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg 3181 tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg 3241 tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg 3301 cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc 3361 agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg 3421 ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc 3481 ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc 3541 gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg 3601 aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgattcgaa atgaccgacc 3661 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt 3721 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca 3781 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa 3841 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 3901 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct 3961 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 4021 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 4081 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc 4141 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg 4201 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc 4261 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt 4321 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc 4381 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa 4441 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc 4501 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg 4561 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc 4621 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc 4681 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca 4741 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact 4801 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg 4861 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt 4921 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct 4981 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga 5041 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa 5101 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac 5161 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga 5221 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc 5281 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca 5341 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta 5401 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg 5461 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc 5521 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg 5581 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt 5641 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt 5701 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata 5761 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc 5821 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac 5881 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa 5941 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct 6001 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat 6061 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc 6121 cacctgacgt cgacattgat tattgactag ttattaatag taatcaatta cggggtcatt 6181 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg 6241 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac 6301 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt 6361 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa 6421 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta 6481 catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt tctgcttcac 6541 tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt tttaattatt 6601 ttgtgcagcg atgggggcgg gggggggggg nggsgcgccr ggcggggsgg ggsggggsgr 6661 ggggsggggs ggggsgrggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg 6721 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg 6781 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg 6841 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc 6901 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct 6961 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg 7021 tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg 7081 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc 7141 ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg 7201 gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc 7261 cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac 7321 ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg 7381 cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag 7441 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga 7501 gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg 7561 ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg 7621 cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg 7681 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg 7741 gcgtgtgacc ggcgg //
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