| 质粒名称: | pCAG-postASAP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 178792 |
| 存储实验室: | Rafael Yuste |
|
载体骨架基本信息 |
|
| 载体名称: | pCAG |
| 载体出品公司: | Clontech |
| 空载体大小: | 6097 bp |
| 完整质粒大小: | 7660bp |
| 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: | FingR.PSD95 and ASAP2f、accelerated sensor of action potentials |
| 物种来源: | Synthetic |
| 插入大小: | 1563 bp |
| 基因突变: | mutations in amino acids L146G, S147T N149R, S150G, H151D, T399R of ASAP2f |
| 启动子: | CAG |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | AATACCACTGAGATCCCCGGGATTACAATGCTCGA |
| 3’测序引物 |
CATGGTGGCGGCTAGGGTTCCGGTGCGGTAGTTGA
(Common Sequencing Primers)
|
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-postASAP was a gift from Rafael Yuste (Addgene plasmid # 178792 ; http://n2t.net/addgene:178792 ; RRID:Addgene_178792) |
| 参考文献部分: | 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 |
买家导航
pCAG-postASAP载体质粒基本信息
pCAG-postASAP质粒图谱载体图谱和pCAG-postASAP载体序列质粒序列多克隆位点信息
pCAG-postASAP质粒载体简介
pCAG-postASAP质粒序列载体序列
LOCUS pCAG-postASAP 7683 bp DNA circular SYN 14-APR-2025
DEFINITION Expresses voltage sensor targeted to dendrites and spines.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7683)
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 7683)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
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-postASAP
FEATURES Location/Qualifiers
source 1..7683
/mol_type="other DNA"
/organism="synthetic DNA construct"
intron 186..1202
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1126..1148
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1210..1229
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
regulatory 1636..1645
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
primer_bind complement(3037..3054)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3043..3267
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3313..3741
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3400..3419)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3610..3631
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3750..3770)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3755..4084
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3935..4070
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3997..4016
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4095..4694
/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(4095..4114)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4591..4611
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 4854..4975
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4891..4910)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4945..4964
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5024..5040)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5024..5040)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5037..5059)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5048..5064
/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(5072..5078,5079..5096,5097..5102))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5117..5138
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5255..5272)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5426..6014)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5506..5525)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(6185..6976,6977..7045))
/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 6808..6827
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7046..7150)
/gene="bla"
/label=AmpR promoter
enhancer 7182..7561
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 gggggggggg gggggggggg gggggggggc gcgcgccggg cggggcgggg cggggcgagg
61 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
121 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
181 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
241 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
301 ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc
361 gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg
421 cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg
481 agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg
541 ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg
601 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc
661 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
721 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
781 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccctggagcg
841 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
901 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
961 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
1021 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
1081 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
1141 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
1201 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattcctcg
1261 aggaattcac tcctcaggtg caggctgcct atcagaaggt ggtggctggt gtggccaatg
1321 ccctggctca caaataccac tgagatcccc gggattacaa tgctcgaagt caaggaagca
1381 tcaccaacca gcatccagat cagctgggtg ctccacttgc gccacgttcg ctactaccgc
1441 atcacctacg gtgaaactgg tggcaatagc cctgtccagg aattcaccgt gcctggcagc
1501 aagtccactg ctaccatcag cggcctgaaa cctggtgtcg actataccat cacggtgtac
1561 gccgtcacga tcttcagcgc ctaccgctcc gcctggccgc cgatctccat caactaccgc
1621 accggaaccc tagccgccac catggagacg actgtgaggt atgaacaggg gtcagagctc
1681 actaaaactt cgagctctcc aacagcagat gagcccacga taaagattga tgatggtcgt
1741 gatgagggta atgaacaaga cagctgttcc aataccatta ggagaaaaat ttccccgttt
1801 gtgatgtcat ttggattcag agtatttgga gttgtgctta tcattgtaga catcatagtg
1861 gtgattgtgg atctggccat cagtgagaag aaaagaggca ttagagagat tcttgaaggt
1921 gtttccctgg ctatagcact cttcttcctt gttgatgttc tcatgagagt gtttgttgaa
1981 ggcttcaaga actatttccg gtccaaactg aatactttgg atgcagtcat agtagtgggc
2041 actctgctaa ttaatatgac ctactccttc tctgacggta cctttagggg cgataacgtg
2101 tatattaccg cggataaaca gaaaaacggc attaaagcga actttaccgt gcgccataac
2161 gtggaagatg gcagcgtgca gctggcggat cattatcagc agaacacccc gattggcgat
2221 ggcccggtgc tgctgccgga taaccattat ctgagcaccc agaccgtgct gagcaaagat
2281 ccgaacgaaa aacgcgatca catggtgctg ctggaatttg tgaccgcagc gggcattaca
2341 cacggcatgg atgaactgta tggcggcacc ggcggcagcg cgagccaggg cgaagaactg
2401 tttaccggcg tggtgccgat tctggtggaa ctggatggcg atgtgaacgg ccataaattt
2461 agcgtgcgcg gcgaaggcga aggcgatgcg accattggca aactgaccct gaaatttatt
2521 tgcaccaccg gcaaactacc ggtgccgtgg ccgaccctgg tgaccacctt aacctatggc
2581 gtgcagtgct ttagccgcta tccggatcat atgaaacgcc atgatttttt taaaagcgcg
2641 atgccggaag gctatgtgca ggaacgcacc attagcttta aagatgatgg caaatataaa
2701 acccgcgcgg tggtgaaatt tgaaggcgat accctggtga accgcattga actgaaaggc
2761 accgatttta aagaagatgg caacattctg gggcataaac tggaatataa cacagatcag
2821 atgccgcaga tggttcgtct tcttcgagtt ctgagaattg ttatcttaat aagaatattt
2881 cgcctggctt cacagaagaa acaacttgaa gtggtaacct aatgaaagct tggtaccgag
2941 ctcggatcca ctagtccagt gtggtggaat tctgcagata tccagcacag tggcggccgc
3001 tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
3061 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
3121 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
3181 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
3241 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
3301 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
3361 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
3421 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
3481 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
3541 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
3601 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
3661 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
3721 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
3781 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
3841 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
3901 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
3961 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
4021 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
4081 caaaaagctc ccgcatgacc gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg
4141 tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca
4201 ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc ttcctcacgc
4261 gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct
4321 ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg
4381 ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc
4441 accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg
4501 gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg
4561 tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag cggctcggct
4621 tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc atgacccgca
4681 agcccggtgc ctgattcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
4741 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
4801 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
4861 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
4921 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
4981 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
5041 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
5101 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
5161 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
5221 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
5281 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
5341 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
5401 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
5461 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
5521 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
5581 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
5641 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
5701 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
5761 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
5821 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
5881 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
5941 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
6001 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
6061 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
6121 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
6181 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
6241 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
6301 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
6361 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
6421 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
6481 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
6541 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
6601 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
6661 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
6721 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
6781 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
6841 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
6901 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
6961 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
7021 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
7081 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
7141 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacattgat tattgactag
7201 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
7261 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
7321 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
7381 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
7441 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
7501 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
7561 ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc
7621 aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg
7681 ggg
//
