| 質粒名稱: | pCAG-VSFP Butterfly 1.2 |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 47978 |
| 存儲實驗室: | Thomas Knopfel |
|
載體骨架基本信息 |
|
| 載體名稱: | pCAG |
| 載體出品公司: | custom |
| 完整質粒大小: | 6860bp |
| 載體類型: | 哺乳細胞表達 |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | XL1 Blue |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | VSFP Butterfly 1.2 |
| 物種來源: | Synthetic |
| 克隆方法: | 限制性內切酶 |
| 5’克隆位點: | NheI (unknown if destroyed) |
| 3’克隆位點: | AflII (unknown if destroyed) |
| 5’測序引物 | atgaggagccgctttgtgaagaaagatggtcattgcaatgttcagtttatcaacgtgatggtgtctaaggg |
| 3’測序引物 |
gaccgccgccgggatcactctcggcatggacgagctgtacaagtaag
(Common Sequencing Primers)
|
|
文獻引用方法 |
|
| 材料和方法部分: | pCAG-VSFP Butterfly 1.2 was a gift from Thomas Knopfel (Addgene plasmid # 47978 ; //n2t.net/addgene:47978 ; RRID:Addgene_47978) |
| 參考文獻部分: | maging neural circuit dynamics with a voltage-sensitive fluorescent protein. Akemann W, Mutoh H, Perron A, Park YK, Iwamoto Y, Knopfel T. J Neurophysiol. 2012 Oct;108(8):2323-37. doi: 10.1152/jn.00452.2012. Epub 2012 Jul 18. 10.1152/jn.00452.2012 PubMed 22815406 |
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pCAG-VSFP Butterfly 1.2載體質粒基本信息
pCAG-VSFP Butterfly 1.2質粒圖譜載體圖譜和pCAG-VSFP Butterfly 1.2載體序列質粒序列多克隆位點信息
pCAG-VSFP Butterfly 1.2質粒載體簡介
pCAG-VSFP Butterfly 1.2質粒序列載體序列
LOCUS pCAG-VSFP Butterfly 1.2 6860 bp DNA circular SYN 14-APR-2025
DEFINITION Genetically encoded voltage indicator.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6860)
AUTHORS Akemann W, Mutoh H, Perron A, Park YK, Iwamoto Y, Knopfel T
TITLE Imaging neural circuit dynamics with a voltage-sensitive fluorescent
protein.
JOURNAL J Neurophysiol. 2012 Oct;108(8):2323-37. doi: 10.1152/jn.00452.2012.
Epub 2012 Jul 18.
PUBMED 22815406
REFERENCE 2 (bases 1 to 6860)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1152/jn.00452.2012"; journalName: "J Neurophysiol"; date:
"2012-10"; volume: "108"; issue: "8"; pages: "2323-37"
COMMENT Sequence Label: pCAG-VSFP Butterfly 1.2
FEATURES Location/Qualifiers
source 1..6860
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..661
/label=chicken beta-actin promoter
intron 662..1670
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1594..1616
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1678..1697
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
CDS join(3051..3053,3054..3056,3057..3770)
/codon_start=1
/product="enhanced YFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
/label=mEYFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(3096..3117)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3357..3376)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3704..3725
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(3884..3903)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 3949..4004
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(4003..4022)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4363..4379)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4363..4379)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4376..4398)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4387..4403
/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(join(4411..4417,4418..4435,4436..4441))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4456..4477
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4536..4731
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 4582..4717
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 4737..4871
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4774..4793)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4828..4847
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4939..4956)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5110..5698)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5190..5209)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(5869..6660,6661..6729))
/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 6492..6511
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6730..6834)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attcgctagc gccaccatga
1741 ggagccgctt tgtgaagaaa gatggtcatt gcaatgttca gtttatcaac gtgatggtgt
1801 ctaagggcga agagctgatt aaggagaaca tgcacatgaa gctgtacatg gagggcaccg
1861 tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac gagggcaccc
1921 agaccatgag aatcaaggcg gtcgagggcg gccctctccc cttcgccttc gacatcctgg
1981 ctaccagctt catgtacggc agcaaaacct tcatcaacca cacccagggc atccccgact
2041 tctttaagca gtccttcccc gagggcttca catgggagag agtcaccaca tacgaagacg
2101 ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc atctacaacg
2161 tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag aaaacactcg
2221 gctgggaggc ctccaccgag accctgtacc ccgctgacgg cggcctggaa ggcagagccg
2281 acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttgaag accacataca
2341 gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg gacagaagac
2401 tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag gtggctgtgg
2461 ccagatactg cgacctccct agcaaactgg ggcacagaga ggaacgaata gatatacccg
2521 agatttcagg tctatggtgg ggcgagaatg aacatggagt ggacgatggg agaatggaga
2581 tacctactac tggtgtaggt cgcgtccagt ttcgtgtccg agcagtgatt gatcatctag
2641 ggatgcgagt ctttggagtc ttcctaattt tcttggacat catcctcatg atcattgatc
2701 tcagtcttcc aggaaaaagt gaatcttcac aatcctttta tgacgggatg gctttggctc
2761 tttcttgtta tttcatgctg gatttaggat taaggatatt tgcctacggg cccaagaatt
2821 tcttcaccaa cccctgggag gttgctgatg gtttgattat cgttgtcaca ttcgtcgtca
2881 cgatatttta cactgtgtta gatgaatacg tgcaagaaac aggagccgat ggtttggggc
2941 agttggttgt gttggcccgt ttgctgcgtg tggttcgatt agcaagaata ttttattccc
3001 accaacaaat gaaggcttca agcagaagaa caatatcaca gtgcggccgc atggtgagca
3061 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
3121 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
3181 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
3241 ccttcggcta cggcctgatg tgcttcgcgc gctaccccga ccacatgaag cagcacgact
3301 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
3361 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
3421 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
3481 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
3541 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
3601 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
3661 accagtccaa actgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
3721 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa ggatccgagc
3781 tcggtaccaa gcttaagtgc ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg
3841 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
3901 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
3961 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
4021 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
4081 ccatatgctg gctgccatga acaaaggtgg ctataaagag gtcatcagta tatgaaacag
4141 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
4201 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
4261 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
4321 tgaagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt
4381 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
4441 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
4501 tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata
4561 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
4621 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
4681 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact
4741 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
4801 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
4861 atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
4921 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
4981 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
5041 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
5101 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
5161 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
5221 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
5281 ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt
5341 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
5401 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
5461 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
5521 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
5581 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
5641 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
5701 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
5761 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
5821 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
5881 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
5941 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
6001 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
6061 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
6121 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
6181 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
6241 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
6301 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
6361 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
6421 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
6481 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
6541 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
6601 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
6661 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
6721 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
6781 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
6841 tccccgaaaa gtgccacctg
//
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