| 质粒名称: | pGP-pcDNA3.1 Puro-CAG-Voltron2 |
|---|---|
| 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 |
|
| 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 |
|
| 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 |
|
| 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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pGP-pcDNA3.1 Puro-CAG-Voltron2载体质粒基本信息
pGP-pcDNA3.1 Puro-CAG-Voltron2质粒图谱载体图谱和pGP-pcDNA3.1 Puro-CAG-Voltron2载体序列质粒序列多克隆位点信息
pGP-pcDNA3.1 Puro-CAG-Voltron2质粒载体简介
pGP-pcDNA3.1 Puro-CAG-Voltron2质粒序列载体序列
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JOURNAL unpublished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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