| 质粒名称: | pN1-GCaMP6m-XC |
|---|---|
| 目录编号: | 111543 |
| 存储实验室: | Xiaodong Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 完整质粒大小: | 5455bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 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: | GCaMP6m-XC/ CBM-GCaMP6m-NES/ GCaMP-X/ NMIQ-GCaMP6m-NES |
| 物种来源: | Synthetic |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | 未知(未破坏) |
| 3’克隆位点: | 未知(未破坏) |
| 5’测序引物 | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | pN1-GCaMP6m-XC was a gift from Xiaodong Liu (Addgene plasmid # 111543 ; http://n2t.net/addgene:111543 ; RRID:Addgene_111543) |
| 参考文献部分: | Improved calcium sensor GCaMP-X overcomes the calcium channel perturbations induced by the calmodulin in GCaMP. Yang Y, Liu N, He Y, Liu Y, Ge L, Zou L, Song S, Xiong W, Liu X. Nat Commun. 2018 Apr 17;9(1):1504. doi: 10.1038/s41467-018-03719-6. 10.1038/s41467-018-03719-6 [pii] PubMed 29666 |
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 pN1-GCaMP6m-XC 5455 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Improved genetic encoded calcium sensor based on GCaMP6m. Also
called GCaMP-X.
ACCESSION .
VERSION .
KEYWORDS pN1-GCaMP6m-XC
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5455)
AUTHORS Yang Y, Liu N, He Y, Liu Y, Ge L, Zou L, Song S, Xiong W, Liu X
TITLE Improved calcium sensor GCaMP-X overcomes the calcium channel
perturbations induced by the calmodulin in GCaMP.
JOURNAL Nat Commun. 2018 Apr 17;9(1):1504. doi: 10.1038/s41467-018-03719-6.
PUBMED 29666364
REFERENCE 2 (bases 1 to 5455)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5455
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 519..539
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 616..625
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 733..2082
/codon_start=1
/product="improved fluorescent protein-based calcium sensor
(Chen et al., 2013)"
/label=GCaMP6m
/translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKGSYRDTEEEIREAFGVFDKDGN
GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
regulatory 829..838
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
primer_bind 1119..1140
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(1246..1267)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1507..1526)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
polyA_signal 2243..2364
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2280..2299)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2334..2353
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(2371..2826)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2508..2529)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2720..2739
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 2853..2957
/gene="bla"
/label=AmpR promoter
promoter 2959..3316
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(2982..3002)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 3167..3302
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3229..3248
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3351..4145
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3405..3424)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4015..4034
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(4333..4352)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4377..4424
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4753..5341
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5242..5261
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atctcgccac catgggcatg gacgagctgt acaagggtac cgctgcaacc
661 aaaattcagg ctgccttccg tggacacata acaaggaaaa agctcaaaga cgagaagaag
721 ggtgctgaat tcatgggttc tcatcatcat catcatcatg gtatggctag catgactggt
781 ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatctcgc caccatggtc
841 gactcatcac gtcgtaagtg gaataagaca ggtcacgcag tcagagctat aggtcggctg
901 agctcactcg agaacgtcta tatcaaggcc gacaagcaga agaacggcat caaggcgaac
961 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag
1021 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcgtgcag
1081 tccaaacttt cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
1141 accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc
1201 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
1261 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac
1321 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
1381 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
1441 cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc
1501 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
1561 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
1621 aagctggagt acaacctgcc ggaccaactg actgaagagc agatcgcaga atttaaagag
1681 gctttctccc tatttgacaa ggacggggat gggacaataa caaccaagga gctggggacg
1741 gtgatgcggt ctctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa
1801 gtagatgccg acggtgacgg cacaatcgac ttccctgagt tcctgacaat gatggcaaga
1861 aaagggagct acagggacac ggaagaagaa attagagaag cgttcggtgt gtttgataag
1921 gatggcaatg gctacatcag tgcagcagag cttcgccacg tgatgacaaa ccttggagag
1981 aagttaacag atgaagaggt tgatgaaatg atcagggaag cagacatcga tggggatggt
2041 caggtaaact acgaagagtt tgtacaaatg atgacagcga agcttgctct taagttggct
2101 ggacttgata ttggttctta agcggccgcg actctagatc ataatcagcc ataccacatt
2161 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
2221 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
2281 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
2341 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
2401 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
2461 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
2521 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
2581 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
2641 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
2701 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
2761 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
2821 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
2881 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
2941 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
3001 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
3061 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
3121 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
3181 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
3241 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
3301 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
3361 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
3421 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
3481 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
3541 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
3601 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
3661 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
3721 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
3781 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
3841 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
3901 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
3961 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
4021 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
4081 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
4141 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
4201 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
4261 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
4321 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
4381 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
4441 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
4501 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
4561 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
4621 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
4681 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
4741 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
4801 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
4861 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
4921 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
4981 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
5041 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
5101 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
5161 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
5221 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
5281 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
5341 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
5401 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
