| 质粒名称: | pcDNA3.1/Puro-CAG-JEDI-1P |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 202607 |
| 存储实验室: | Francois St-Pierre |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1/Puro-CAG |
| 空载体大小: | 6088 bp |
| 完整质粒大小: | 7380 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | Puromycin |
| 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: | JEDI-1P |
| 物种来源: | Synthetic |
| 插入大小: | 1284bp |
| 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 |
|
| 材料和方法部分: | pcDNA3.1/Puro-CAG-JEDI-1P was a gift from Francois St-Pierre (Addgene plasmid # 202607 ; http://n2t.net/addgene:202607 ; RRID:Addgene_202607) |
| 参考文献部分: | Widefield imaging of rapid pan-cortical voltage dynamics with an indicator evolved for one-photon microscopy. Lu X, Wang Y, Liu Z, Gou Y, Jaeger D, St-Pierre F. Nat Commun. 2023 Oct 12;14(1):6423. doi: 10.1038/s41467-023-41975-3. 10.1038/s41467-023-41975-3 PubMed 37828037 |
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-1P 7380 bp ds-DNA circular SYN 22-JUN-2023
DEFINITION Genetically encoded voltage indicator (GEVI) JEDI-1P, expressed
under strong mammalian promoter (CAG).
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1/Puro-CAG-JEDI-1P
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7380)
TITLE Detecting rapid pan-cortical voltage dynamics in vivo with a
brighter and faster voltage indicator
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7380)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jun 22, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7380
/organism="synthetic DNA construct"
/mol_type="other DNA"
regulatory 112..121
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind complement(1516..1533)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1522..1746
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 1792..2220
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(1879..1898)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2089..2110
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2229..2249)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2234..2563
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2414..2549
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2476..2495
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2574..3173
/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(2574..2593)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3070..3090
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 3333..3454
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3370..3389)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3424..3443
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3503..3519)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3503..3519)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3516..3538)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3527..3543
/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(3551..3581)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3596..3617
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3734..3751)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3905..4493)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3985..4004)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4664..5524)
/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 5287..5306
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5525..5629)
/gene="bla"
/label=AmpR promoter
enhancer 5661..6040
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
intron 6316..7332
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 7256..7278
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 7340..7359
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
ORIGIN
1 gaattcctcg aggaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt
61 gtggccaatg ccctggctca caaataccac tgagatctgc atgagctagc cgccaccatg
121 gagacgactg tgaggtatga acaggggtca gagctcacta aaacttcgag ctctccaaca
181 gcagatgagc ccacgataaa gattgatgat ggtcgtgatg agggtaatga acaagacagc
241 tgttccaata ccattaggag aaaaatttcc ccgtttgtga tgtcatttgg attcagagta
301 tttggagttg tgcttatcat tgtagacatc atagtggtga ttgtggatct ggccatcagt
361 gagaagaaaa gaggcattag agagattctt gaaggtgttt ccctggctat agcactcttc
421 ttccttgttg atgttctcat gagagtgttt gttgaaggct tcaagaacta tttccggtcc
481 aaactggtta ctttggatgc agtcatagta gtgggcactc tgctaattaa tatgacctac
541 tccttctctg accttgctgc ctttaacagc gagaacgtgt atattaccgc ggataaacag
601 aaaaacggca ttaaagcgaa ctttaccgtg cgccataacg tggaagatgg cagcgtgcag
661 ctggcggatc attatcagca gaacaccccg attggcgatg gcccggtgct gctgccggat
721 aaccattatc tgagcaccca gaccgtgctg agcaaagatc cgaacgaaaa acgcgatcac
781 atggtgctgc tggaatttgt gaccgcagcg ggcattacac acggcatgga tgaactgtat
841 ggcggcaccg gcggcagcgc gagccagggc gaagaactgt ttaccggcgt ggtgccgatt
901 ctggtggaac tggatggcga tgtgaacggc cataaattta gcgtgcgcgg cgaaggcgaa
961 ggcgatgcga ccattggcaa actgaccctg aaatttattt gcaccaccgg caaactaccg
1021 gtgccgtggc cgaccctggt gaccacctta acctatggcg tgcagtgctt tagccgctat
1081 ccggatcata tgaaacgcca tgattttttt aaaagcgcga tgccggaagg ctatgtgcag
1141 gaacgcacca ttagctttaa agatgatggc aaatataaaa cccgcgcggt ggtgaaattt
1201 gaaggcgata ccctggtgaa ccgcattgaa ctgaaaggca ccgattttaa agaagatggc
1261 aacattctgg ggcataaact ggaatataac acagatcaga tgccgcacat ggttactctt
1321 cttcgagttc tgaaaattgt tatcttaata agaatatttc gcctggcttc acagaagaaa
1381 caacttgaag tggtaaccta atgaaagctt ggtaccgagc tcggatccac tagtccagtg
1441 tggtggaatt ctgcagatat ccagcacagt ggcggccgct cgagtctaga gggcccgttt
1501 aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
1561 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
1621 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
1681 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
1741 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
1801 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
1861 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
1921 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
1981 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
2041 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
2101 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
2161 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
2221 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
2281 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
2341 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
2401 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
2461 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
2521 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgcatgaccg
2581 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
2641 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
2701 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
2761 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
2821 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
2881 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
2941 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
3001 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
3061 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
3121 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgattcgaaa
3181 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
3241 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
3301 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
3361 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
3421 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
3481 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
3541 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
3601 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
3661 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
3721 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
3781 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
3841 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
3901 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
3961 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
4021 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
4081 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
4141 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
4201 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
4261 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
4321 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
4381 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
4441 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
4501 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
4561 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
4621 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
4681 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
4741 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
4801 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
4861 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
4921 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
4981 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
5041 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
5101 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
5161 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
5221 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
5281 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
5341 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
5401 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
5461 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
5521 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
5581 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
5641 gaaaagtgcc acctgacgtc gacattgatt attgactagt tattaatagt aatcaattac
5701 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
5761 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
5821 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
5881 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
5941 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
6001 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
6061 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
6121 ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgccagg cggggcgggg
6181 cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc
6241 gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga
6301 agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc
6361 gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg
6421 gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt
6481 ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc
6541 tcggggggtg cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc
6601 ggcggctgtg agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag
6661 gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc
6721 tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg
6781 caaccccccc tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg
6841 ctccgtacgg ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg
6901 gtgccgggcg gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc
6961 ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa
7021 tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg
7081 aggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag
7141 gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag
7201 cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc
7261 ggcttctggc gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc
7321 tttttcctac agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa
//
