| 质粒名称: | pGP-CMV-GCaMP6s |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 40753 |
| 存储实验室: | Douglas Kim |
|
载体骨架基本信息 |
|
| 载体名称: | pEGFP-N1 |
| 载体出品公司: | Clontech |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 3955 bp |
| 完整质粒大小: | 5308 bp |
|
载体类型: |
哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | GCaMP6s |
| 物种来源: | R. norvegicus (rat), G. gallus (chicken); A. victoria (jellyfish) |
| 插入大小: | 1353 bp |
| 基因又名: | GCaMP3-K78H T302L R303P D380Y T381R S383T R392G |
| 启动子 | CMV |
| 标签/融合蛋白: |
6xHis (N terminal on insert)
T7 epitope (N terminal on insert)
Xpress tag (N terminal on insert)
|
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BglII(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV Forward |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pGP-CMV-GCaMP6s was a gift from Douglas Kim (爱迪基因 plasmid # 40753) |
| 参考文献部分: |
Ultrasensitive fluorescent proteins for imaging neuronal activity. Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A, Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K, Kim DS. Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354. 10.1038/nature12354 PubMed 23868258 |
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 pGP-CMV-GCaMP6s 5308 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Mammalian expression of ultrasensitive protein calcium sensor.
ACCESSION .
VERSION .
KEYWORDS pGP-CMV-GCaMP6s
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5308)
AUTHORS Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A,
Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K,
Kim DS
TITLE Ultrasensitive fluorescent proteins for imaging neuronal activity.
JOURNAL Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354.
PUBMED 23868258
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pGP-CMV-GCaMP6s|
FEATURES Location/Qualifiers
source 1..5308
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 191..211
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter complement(236..340)
/gene="bla"
/label=AmpR promoter
rep_origin 367..822
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(454..473)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 664..685
/label=F1ori-F
/note="F1 origin, forward primer"
polyA_signal 829..950
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(840..859)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 894..913
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(1072..2424)
/codon_start=1
/product="improved fluorescent protein-based calcium sensor
(Chen et al., 2013)"
/label=GCaMP6s
/translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
primer_bind 1631..1650
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 1890..1911
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2017..2038)
/label=EGFP-C
/note="EGFP, forward primer"
regulatory 2319..2328
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 2421..2430
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter complement(2478..2681)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(2507..2527)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 2682..2985
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
rep_origin complement(3160..3748)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3240..3259)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
polyA_signal 4077..4124
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
primer_bind 4149..4168
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
CDS complement(4356..5150)
/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(4467..4486)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 5077..5096
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter complement(5185..234)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5199..26
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(5253..5272)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
ORIGIN
1 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
61 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
121 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
181 gactttccac accctaactg acacacattc cacagctggt tctttccgcc tcaggactct
241 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
301 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
361 cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
421 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
481 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
541 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
601 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
661 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
721 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
781 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttacgcctta agatacattg
841 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
901 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
961 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
1021 aaaacctcta caaatgtggt atggctgatt atgatctaga gtcgcggccg ctcacttcgc
1081 tgtcatcatt tgtacaaact cttcgtagtt tacctgacca tccccatcga tgtctgcttc
1141 cctgatcatt tcatcaacct cttcatctgt taacttctct ccaaggtttg tcatcacgtg
1201 gcgaagctct gctgcactga tgtagccatt gccatcctta tcaaacacac cgaacgcttc
1261 tctaatttct tcttccgtgt ccctgtattt catttttctt gccatcattg tcaggaactc
1321 agggaagtcg attgtgccgt caccgtcggc atctacttca ttgatcatgt cctgcagctc
1381 tgcttctgtg gggttctgcc ccagagaccg catcaccgtc cccagctcct tggttgttat
1441 tgtcccatcc ccgtccttgt caaataggga gaaagcctct ttaaattctg cgatctgctc
1501 ttcagtcagt tggtccggca ggttgtactc cagcttgtgc cccaggatgt tgccgtcctc
1561 cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac
1621 ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggatgta
1681 gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcggct
1741 gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca cgggcagctt
1801 gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcaccctc
1861 gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac
1921 caccccggtg aacagctcct cgcccttgct caccatgctc cctccggtac cgcccttgta
1981 cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg
2041 atcgcgcttc tcgttggggt ctttcgaaag tttggactgc acgctcaggt agtggttgtc
2101 gggcagcagc acggggccgt cgccgatggg ggtgttctgc tggtagtggt aggcgagctg
2161 cacgccgccg tcctcgatgt tgtggcggat gtggaagttc gccttgatgc cgttcttctg
2221 cttgtcggcc ttgatataga cgttctcgag tgagctcagc cgacctatag ctctgactgc
2281 gtgacctgtc ttattccact tacgacgtga tgagtcgacc atggtggcga gatccttatc
2341 gtcatcgtcg tacagatccc gacccatttg ctgtccacca gtcatgctag ccataccatg
2401 atgatgatga tgatgagaac ccatggtggc gagatctgag tccggtagcg ctagcggatc
2461 tgacggttca ctaaaccagc tctgcttata tagacctccc accgtacacg cctaccgccc
2521 atttgcgtca atggggcgga gttgttacga cattttggaa agtcccgttg attttggtgc
2581 caaaacaaac tcccattgac gtcaatgggg tggagacttg gaaatccccg tgagtcaaac
2641 cgctatccac gcccattgat gtactgccaa aaccgcatca ccatggtaat agcgatgact
2701 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
2761 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
2821 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
2881 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg gcgggggtcg
2941 ttgggcggtc agccaggcgg gccatttacc gtaagttatg taacgcggaa ctccatatat
3001 gggctatgaa ctaatgaccc cgtaattgat tactattaat aactaatgca tggcggtaat
3061 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
3121 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
3181 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
3241 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
3301 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
3361 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
3421 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
3481 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
3541 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
3601 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
3661 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
3721 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
3781 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
3841 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
3901 gtaacctgag gctatggcag ggcctgccgc cccgacgttg gctgcgagcc ctgggccttc
3961 acccgaactt ggggggtggg gtggggaaaa ggaagaaacg cgggcgtatt ggccccaatg
4021 gggtctcggt ggggtatcga cagagtgcca gccctgggac cgaaccccgc gtttatgaac
4081 aaacgaccca acaccgtgcg ttttattctg tctttttatt gccgtcatag cgcgggttcc
4141 ttccggtatt gtctccttcc gtgtttcagt tagcctcccc ctagggtggg cgaagaactc
4201 cagcatgaga tccccgcgct ggaggatcat ccagccggcg tcccggaaaa cgattccgaa
4261 gcccaacctt tcatagaagg cggcggtgga atcgaaatct cgtgatggca ggttgggcgt
4321 cgcttggtcg gtcatttcga accccagagt cccgctcaga agaactcgtc aagaaggcga
4381 tagaaggcga tgcgctgcga atcgggagcg gcgataccgt aaagcacgag gaagcggtca
4441 gcccattcgc cgccaagctc ttcagcaata tcacgggtag ccaacgctat gtcctgatag
4501 cggtccgcca cacccagccg gccacagtcg atgaatccag aaaagcggcc attttccacc
4561 atgatattcg gcaagcaggc atcgccatgg gtcacgacga gatcctcgcc gtcgggcatg
4621 ctcgccttga gcctggcgaa cagttcggct ggcgcgagcc cctgatgctc ttcgtccaga
4681 tcatcctgat cgacaagacc ggcttccatc cgagtacgtg ctcgctcgat gcgatgtttc
4741 gcttggtggt cgaatgggca ggtagccgga tcaagcgtat gcagccgccg cattgcatca
4801 gccatgatgg atactttctc ggcaggagca aggtgagatg acaggagatc ctgccccggc
4861 acttcgccca atagcagcca gtcccttccc gcttcagtga caacgtcgag cacagctgcg
4921 caaggaacgc ccgtcgtggc cagccacgat agccgcgctg cctcgtcttg cagttcattc
4981 agggcaccgg acaggtcggt cttgacaaaa agaaccgggc gcccctgcgc tgacagccgg
5041 aacacggcgg catcagagca gccgattgtc tgttgtgccc agtcatagcc gaatagcctc
5101 tccacccaag cggccggaga acctgcgtgc aatccatctt gttcaatcat gcgaaacgat
5161 cctcatcctg tctcttgatc gatctttgca aaagcctagg cctccaaaaa agcctcctca
5221 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
5281 tagtcagcca tggggcggag aatgggcg
//
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