| 质粒名称: | 5HT6-mCherry-G-GECO1.0 |
|---|---|
| 出品公司: | Clontech |
| 目录编号: | 47500 |
| 存储实验室: | Takanari Inoue |
|
载体骨架基本信息 |
|
| 载体名称: | pmCherry-C1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 4700bp |
| 完整质粒大小: | 7308 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | G-GECO1.0 |
| 物种来源: | Synthetic |
| 插入大小: | 1250 bp |
| 标签/融合蛋白: |
5HT6 (N terminal on insert)
mCherry (N terminal on insert)
|
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI(未破坏) |
| 3’克隆位点: | EcoRI(未破坏) |
| 5’测序引物 | GAACAGTACGAACGCGCC |
|
文献引用方法 |
|
| 材料和方法部分: | |
| 参考文献部分: | |
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 5HT6-mCherry-G-GECO1.0 7308 bp ds-DNA circular SYN 27-SEP-2021
DEFINITION Fluorescent reporter for Ca2+ inside primary cilia with mCherry
reference.
ACCESSION .
VERSION .
KEYWORDS 5HT6-mCherry-G-GECO1.0
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7308)
AUTHORS Su S, Phua SC, Derose R, Chiba S, Narita K, Kalugin PN, Katada T,
Kontani K, Takeda S, Inoue T
TITLE Genetically encoded calcium indicator illuminates calcium dynamics
in primary cilia.
JOURNAL Nat Methods. 2013 Sep 22. doi: 10.1038/nmeth.2647.
PUBMED 24056873
REFERENCE 2 (bases 1 to 7308)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 27, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7308
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 305..508
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 459..479
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 537..546
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
regulatory 1893..1902
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1899..2606
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(2044..2062)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 2312..2331
/label=mCherry-F
/note="mCherry, forward primer"
CDS 2640..2696
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKTGHAVRAIGRLSS"
primer_bind 2909..2930
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(3297..3316)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
CDS 3432..3875
/codon_start=1
/product="calcium-binding protein"
/label=calmodulin
/translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYIGAAEL
RHVMTNLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
polyA_signal 4036..4157
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4073..4092)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4127..4146
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin complement(4164..4619)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4301..4322)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4513..4532
/label=F1ori-R
/note="F1 origin, reverse primer"
promoter 4646..4750
/gene="bla"
/label=AmpR promoter
promoter 4752..5109
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
primer_bind complement(4775..4795)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin 4960..5095
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5022..5041
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5144..5938
/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(5198..5217)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5808..5827
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind complement(6126..6145)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 6170..6217
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6546..7134
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7035..7054
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
541 ccatggttcc agagcccggc cctgtcaaca gtagcacccc agcctggggc cccgggccac
601 cgccagcccc agggggcagc ggctgggtgg ccgctgcgct gtgcgtggtc atcgtgctga
661 cggcagccgc caattcgctg ctgatcgcgc tcatctgcac gcagcccgcg ctgcgcaaca
721 cgtctaactt cttcctggtg tcgctcttca cgtcggacct gatggtggga ttggtggtga
781 tgcccccagc catgctgaac gcgctgtatg ggcgctgggt actagctcgc ggcctctgtc
841 tgctttggac cgccttcgac gtgatgtgct gcagcgcctc cattctcaac ctctgcctca
901 tcagcctgga ccgctacctg ctcatcctct cgccgctgcg ctacaagctg cgcatgacag
961 ccccgcgtgc cctggcgctc atcctgggtg cctggagcct cgccgcgctg gcctccttcc
1021 tgcccctctt gctgggctgg catgaactgg gcaaagctcg aacatctgcc ccgggccagt
1081 gccgcctatt ggccagcctg ccttatgtcc tcgtggcgtc cggcgtcacc tttttcctgc
1141 cttcgggtgc catctgcttc acctactgca ggatcttgct ggctgcccgc aagcaggcgg
1201 tgcaagtggc ctcgctcacc acgggcacgg ctacggctgg ccaggccttg gaaaccttgc
1261 aggtgcccag gacaccacgc ccagggatgg agtctgctga cagtaggcgt ctgaccacca
1321 agcatagcag gaaggccttg aaggccagcc tgaccctcgg catcctgctg agcatgttct
1381 ttgtcacctg gctgcccttc tttgtggcca gcatagctca ggccgtatgt gactgcatct
1441 ctccaggtct cttcgatgtc ctcacatggc tgggatactg taatagcacc atgaacccta
1501 tcatctatcc cctcttcatg cgggacttca agagggccct gggcaggttc gtgccgtgtg
1561 tccactgtcc cccggagcac cgggccagcc ccgcctcccc ctccatgtgg acctctcaca
1621 gtggtgccag gccaggcctc agcctgcagc aggtgctgcc cctgcctctg ccacccaact
1681 cagattcaga ctcagcttca gggggcacct cgggcctgca gctcacagcc cagcttttgc
1741 tgcctggaga ggcgacccgg gaccccccgc cacccaccag ggcccctact gtggtcaact
1801 tcttcgtcac agactctgtg gagcctgaga tacggcagca tccacttggt tcccccatga
1861 acagtgctgg tggtagcgca ggaggaccgg tcgccaccat ggtaagcaag ggcgaggagg
1921 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
1981 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
2041 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
2101 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
2161 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
2221 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
2281 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
2341 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
2401 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
2461 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
2521 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
2581 ccggcggcat ggacgagctg tacaagtccg gactcagatc caccatggtc gactcatcac
2641 gtcgtaagtg gaataagaca ggtcacgcag tcagagctat aggtcggctg agctcactcg
2701 agaacgtcta tatcaaggcc gacaagcaga agaacggcat caaggcgaac ttcaagatcc
2761 gccacaacat cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag aacaccccca
2821 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcgtgcag tccatacttt
2881 cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
2941 ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc atggtgagca
3001 agggcgagga gctgttcacc ggggtggtgc ccatccaggt cgagctggac ggcgacgtaa
3061 acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac ggcaagctga
3121 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
3181 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
3241 tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc ttcaaggacg
3301 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
3361 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
3421 acaacacgcg tgaccaactg actgaagagc agatcgcaga atttaaagag gctttctccc
3481 tatttgacaa ggacggggat gggacgataa caaccaagga gctggggacg gtgatgcggt
3541 ctctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa gtagatgccg
3601 acggtgacgg cacaatcgac ttccctgagt tcctgacaat gatggcaaga aaaatgaaag
3661 acacagacag tgaagaagaa attagagaag cgttccgtgt gtttgataag gacggcaatg
3721 gctacatcgg cgcagcagag cttcgccacg tgatgacaaa ccttggagag aagttaacag
3781 atgaagaggt tgatgaaatg atcagggtag cagacatcga tggggatggt caggtaaact
3841 acgaagagtt tgtacaaatg atgacagcga agtaagaatt ctgcagtcga cggtaccgcg
3901 ggcccgggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
3961 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
4021 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
4081 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
4141 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
4201 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
4261 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
4321 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
4381 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
4441 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
4501 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
4561 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
4621 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
4681 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
4741 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
4801 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
4861 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
4921 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
4981 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
5041 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
5101 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
5161 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
5221 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
5281 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
5341 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
5401 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
5461 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
5521 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
5581 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
5641 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
5701 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
5761 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
5821 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
5881 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
5941 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
6001 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
6061 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
6121 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
6181 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
6241 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
6301 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
6361 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
6421 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
6481 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
6541 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
6601 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
6661 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
6721 cacttcaaga actctgtatc accgcctaca tacctcgctc tgctaatcct gttaccagtg
6781 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
6841 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
6901 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
6961 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
7021 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
7081 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
7141 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
7201 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
7261 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccg
//
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