| 質粒名稱: | pcDNA3.1/Puro-CAG-JEDI-2P |
|---|---|
| 出品公司： | addgene |
| 目錄編號: | 179458 |
| 存儲實驗室: | Francois St-Pierre |
|
載體骨架基本信息 |
|
| 載體名稱: | pcDNA3.1/Puro-CAG |
| 空載體大小: | 6095 bp |
| 完整質粒大小: | 7383 bp |
| 載體類型: | 哺乳細胞表達 |
| 篩選標記: | 新霉素（G418） |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | JEDI-2P / JEDI2P |
| 物種來源: | Synthetic |
| 插入大小: | 1287 bp |
| 啟動子: | CAG |
| 克隆方法: | 限制性內切酶 |
| 5’克隆位點: | NheI(未破壞) |
| 3’克隆位點: | HindIII (未破壞) |
| 5’測序引物 | AAGGTGGTGGCTGGTGTGGC |
| 3’測序引物 | TAGAAGGCACAGTCGAGG |
|
文獻引用方法 |
|
| 材料和方法部分: | pcDNA3.1/Puro-CAG-JEDI-2P was a gift from Francois St-Pierre (Addgene plasmid # 179458 ; //n2t.net/addgene:179458 ; RRID:Addgene_179458) |
| 參考文獻部分: | 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 |
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pcDNA3.1/Puro-CAG-JEDI-2P載體質粒基本信息
pcDNA3.1/Puro-CAG-JEDI-2P質粒圖譜載體圖譜和pcDNA3.1/Puro-CAG-JEDI-2P載體序列質粒序列多克隆位點信息
pcDNA3.1/Puro-CAG-JEDI-2P質粒載體簡介
pcDNA3.1/Puro-CAG-JEDI-2P質粒序列載體序列
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JOURNAL Unpublished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
//gt12365.cn
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
//