| 質粒名稱: | pCAG-RFP-p2a-postASAP |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 178793 |
| 存儲實驗室: | Rafael Yuste |
|
載體骨架基本信息 |
|
| 載體名稱: | pCAG |
| 載體出品公司: | Clontech |
| 空載體大小: | 5466 bp |
| 完整質粒大小: | 7821bp |
| 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: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | FingR.PSD95, ASAP2f and TagRFP、accelerated sensor of action potentials 2f |
| 物種來源: | Synthetic |
| 插入大小: | 2355bp |
| 基因突變: | mutations in amino acids L146G, S147T N149R, S150G, H151D, T399R of ASAP2f |
| 啟動子: | CAG |
| 標簽/融合蛋白: | TagRFP |
| 克隆方法: | Gibson Cloning |
| 5’測序引物 | taccggtaggccaccatggtgtctaagggcgaagagct |
| 3’測序引物 |
cttgacttcgagcatgggaccggggttttcttcca
(Common Sequencing Primers)
|
|
文獻引用方法 |
|
| 材料和方法部分: | pCAG-RFP-p2a-postASAP was a gift from Rafael Yuste (Addgene plasmid # 178793 ; //n2t.net/addgene:178793 ; RRID:Addgene_178793) |
| 參考文獻部分: | Voltage compartmentalization in dendritic spines in vivo. Cornejo VH, Ofer N, Yuste R. Science. 2022 Jan 7;375(6576):82-86. doi: 10.1126/science.abg0501. Epub 2021 Nov 11. 10.1126/science.abg0501 PubMed 34762487 |
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pCAG-RFP-p2a-postASAP載體質粒基本信息
pCAG-RFP-p2a-postASAP質粒圖譜載體圖譜和pCAG-RFP-p2a-postASAP載體序列質粒序列多克隆位點信息
pCAG-RFP-p2a-postASAP質粒載體簡介
pCAG-RFP-p2a-postASAP質粒序列載體序列
LOCUS pCAG-RFP-p2a-postASAP 7829 bp DNA circular SYN 14-APR-2025
DEFINITION Expresses voltage sensor targeted to dendrites and spines along with
RFP.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7829)
AUTHORS Cornejo VH, Ofer N, Yuste R
TITLE Voltage compartmentalization in dendritic spines in vivo.
JOURNAL Science. 2022 Jan 7;375(6576):82-86. doi: 10.1126/science.abg0501.
Epub 2021 Nov 11.
PUBMED 34762487
REFERENCE 2 (bases 1 to 7829)
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:
"10.1126/science.abg0501"; journalName: "Science"; date: "2022-01-7-
7"; volume: "375"; issue: "6576"; pages: "82-86"
COMMENT Sequence Label: pCAG-RFP-p2a-postASAP
FEATURES Location/Qualifiers
source 1..7829
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 5..384
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 386..660
/label=chicken beta-actin promoter
intron 661..1677
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1601..1623
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1685..1704
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1765..1774
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1771..2481
/codon_start=1
/product="monomeric derivative of red fluorescent protein
from Entacmaea quadricolor (Merzlyak et al., 2007)"
/label=TagRFP
/note="mammalian codon-optimized"
/translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
ARYCDLPSKLGHKLN"
CDS 2506..2562
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
regulatory 2839..2848
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
misc_feature 4150..4738
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(4203..4223)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(4621..4632)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(4851..4870)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 4916..4971
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(4970..4989)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(5332..5348)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5332..5348)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5345..5367)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5356..5372
/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(5380..5386,5387..5404,5405..5410))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5425..5446
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5505..5700
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 5551..5686
/label=SV40 ori
/note="SV40 origin of replication"
polyA_signal 5706..5840
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5743..5762)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5797..5816
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5908..5925)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6079..6667)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6159..6178)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(6838..7629,7630..7698))
/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 7461..7480
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7699..7803)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
481 gcgatggggg cggggggggg ggggcgcgcg ccaggcgggg cggggsgggg sgrggggsgg
541 ggsggggsgr ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc gggcggggcg
1261 gggccgcctc gggccggggr gggctcgggg grggggcgcg gcggcccccg gagcgccggc
1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt tctagagcgg
1741 taccggatcc ccgggtaccg gtaggccacc atggtgtcta agggcgaaga gctgattaag
1801 gagaacatgc acatgaagct gtacatggag ggcaccgtga acaaccacca cttcaagtgc
1861 acatccgagg gcgaaggcaa gccctacgag ggcacccaga ccatgagaat caaggtggtc
1921 gagggcggcc ctctcccctt cgccttcgac atcctggcta ccagcttcat gtacggcagc
1981 agaaccttca tcaaccacac ccagggcatc cccgacttct ttaagcagtc cttccctgag
2041 ggcttcacat gggagagagt caccacatac gaagacgggg gcgtgctgac cgctacccag
2101 gacaccagcc tccaggacgg ctgcctcatc tacaacgtca agatcagagg ggtgaacttc
2161 ccatccaacg gccctgtgat gcagaagaaa acactcggct gggaggccaa caccgagatg
2221 ctgtaccccg ctgacggcgg cctggaaggc agaagcgaca tggccctgaa gctcgtgggc
2281 gggggccacc tgatctgcaa cttcaagacc acatacagat ccaagaaacc cgctaagaac
2341 ctcaagatgc ccggcgtcta ctatgtggac cacagactgg aaagaatcaa ggaggccgac
2401 aaagagacct acgtcgagca gcacgaggtg gctgtggcca gatactgcga cctccctagc
2461 aaactggggc acaaacttaa tggcgcgccc gcgcccggct ccggagccac gaacttctct
2521 ctgttaaagc aagcaggaga cgtggaagaa aaccccggtc ccatgctcga agtcaaggaa
2581 gcatcaccaa ccagcatcca gatcagctgg gtgctccact tgcgccacgt tcgctactac
2641 cgcatcacct acggtgaaac tggtggcaat agccctgtcc aggaattcac cgtgcctggc
2701 agcaagtcca ctgctaccat cagcggcctg aaacctggtg tcgactatac catcacggtg
2761 tacgccgtca cgatcttcag cgcctaccgc tccgcctggc cgccgatctc catcaactac
2821 cgcaccggaa ccctagccgc caccatggag acgactgtga ggtatgaaca ggggtcagag
2881 ctcactaaaa cttcgagctc tccaacagca gatgagccca cgataaagat tgatgatggt
2941 cgtgatgagg gtaatgaaca agacagctgt tccaatacca ttaggagaaa aatttccccg
3001 tttgtgatgt catttggatt cagagtattt ggagttgtgc ttatcattgt agacatcata
3061 gtggtgattg tggatctggc catcagtgag aagaaaagag gcattagaga gattcttgaa
3121 ggtgtttccc tggctatagc actcttcttc cttgttgatg ttctcatgag agtgtttgtt
3181 gaaggcttca agaactattt ccggtccaaa ctgaatactt tggatgcagt catagtagtg
3241 ggcactctgc taattaatat gacctactcc ttctctgacg gtacctttag gggcgataac
3301 gtgtatatta ccgcggataa acagaaaaac ggcattaaag cgaactttac cgtgcgccat
3361 aacgtggaag atggcagcgt gcagctggcg gatcattatc agcagaacac cccgattggc
3421 gatggcccgg tgctgctgcc ggataaccat tatctgagca cccagaccgt gctgagcaaa
3481 gatccgaacg aaaaacgcga tcacatggtg ctgctggaat ttgtgaccgc agcgggcatt
3541 acacacggca tggatgaact gtatggcggc accggcggca gcgcgagcca gggcgaagaa
3601 ctgtttaccg gcgtggtgcc gattctggtg gaactggatg gcgatgtgaa cggccataaa
3661 tttagcgtgc gcggcgaagg cgaaggcgat gcgaccattg gcaaactgac cctgaaattt
3721 atttgcacca ccggcaaact accggtgccg tggccgaccc tggtgaccac cttaacctat
3781 ggcgtgcagt gctttagccg ctatccggat catatgaaac gccatgattt ttttaaaagc
3841 gcgatgccgg aaggctatgt gcaggaacgc accattagct ttaaagatga tggcaaatat
3901 aaaacccgcg cggtggtgaa atttgaaggc gataccctgg tgaaccgcat tgaactgaaa
3961 ggcaccgatt ttaaagaaga tggcaacatt ctggggcata aactggaata taacacagat
4021 cagatgccgc agatggttcg tcttcttcga gttctgagaa ttgttatctt aataagaata
4081 tttcgcctgg cttcacagaa gaaacaactt gaagtggtaa cctaggaatt cgatatcaag
4141 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
4201 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
4261 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
4321 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
4381 acccccactg gttggggaat tgccaccacc tgtcagctcc tttccggaac tttcgctttc
4441 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
4501 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
4561 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
4621 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
4681 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
4741 cgataccgag cgctgctcga gagatctggc cgcactcctc aggtgcaggc tgcctatcag
4801 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
4861 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
4921 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
4981 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
5041 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat
5101 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag
5161 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta
5221 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct
5281 cttctcttat ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct
5341 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
5401 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
5461 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
5521 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
5581 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
5641 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
5701 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
5761 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
5821 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
5881 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
5941 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
6001 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
6061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
6121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
6181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
6241 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
6301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
6361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
6421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
6481 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
6541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
6601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
6661 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
6721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
6781 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
6841 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
6901 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
6961 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
7021 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
7081 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
7141 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
7201 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
7261 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
7321 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
7381 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
7441 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
7501 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
7561 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
7621 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
7681 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
7741 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
7801 gcgcacattt ccccgaaaag tgccacctg
//
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