| 质粒名称: | 5HT6-G-GECO1.0 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 47499 |
| 存储实验室: | Takanari Inoue |
|
载体骨架基本信息 |
|
| 载体名称: | Customized |
| 空载体大小: | 4450 bp |
| 完整质粒大小: | 5807bp |
| 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: | 未知 |
|
插入基因信息 |
|
| 插入基因名称: |
5-hydroxytriptamine receptor isoform 6 / 5HT6
|
| 物种来源: | M. musculus (mouse) |
| 插入大小: | 1350bp |
| GenBank编号 | NM_021358 |
| 标签/融合蛋白: | G-GECO1.0 (C terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | BamHI (未破坏) |
| 3’克隆位点: | BamHI (未破坏) |
| 5’测序引物 | CMV-F |
| 3’测序引物 | SP6 |
|
文献引用方法 |
|
| 材料和方法部分: | 5HT6-G-GECO1.0 was a gift from Takanari Inoue (Addgene plasmid # 47499 ; http://n2t.net/addgene:47499 ; RRID:Addgene_47499) |
| 参考文献部分: | Genetically encoded calcium indicator illuminates calcium dynamics in primary cilia. Su S, Phua SC, Derose R, Chiba S, Narita K, Kalugin PN, Katada T, Kontani K, Takeda S, Inoue T. Nat Methods. 2013 Sep 22. doi: 10.1038/nmeth.2647. 10.1038/nmeth.2647 PubMed 24056873 |
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-G-GECO1.0 5807 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Fluorescent reporter for Ca2+ inside primary cilia.
ACCESSION .
VERSION .
KEYWORDS 5HT6-G-GECO1.0
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5807)
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 5807)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..5807
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 629..648
/label=T7
/note="T7 promoter, forward primer"
promoter 629..647
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 679..688
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2053..2109
/codon_start=1
/product="calmodulin-binding peptide from avian smooth
muscle myosin light chain kinase"
/label=calmodulin-binding peptide
/translation="RRKWNKTGHAVRAIGRLSS"
primer_bind 2322..2343
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(2710..2729)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
CDS 2845..3288
/codon_start=1
/product="calcium-binding protein"
/label=calmodulin
/translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYIGAAEL
RHVMTNLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
promoter complement(3349..3367)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(3350..3367)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(3387..3404)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3393..3617
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin complement(3818..4406)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3898..3917)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4577..5437)
/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 5200..5219
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5438..5542)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5617..5636)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
661 ggtaccgagc tcggatccgc caccatggtt ccagagcccg gccctgtcaa cagtagcacc
721 ccagcctggg gccccgggcc accgccagcc ccagggggca gcggctgggt ggccgctgcg
781 ctgtgcgtgg tcatcgtgct gacggcagcc gccaattcgc tgctgatcgc gctcatctgc
841 acgcagcccg cgctgcgcaa cacgtctaac ttcttcctgg tgtcgctctt cacgtcggac
901 ctgatggtgg gattggtggt gatgccccca gccatgctga acgcgctgta tgggcgctgg
961 gtactagctc gcggcctctg tctgctttgg accgccttcg acgtgatgtg ctgcagcgcc
1021 tccattctca acctctgcct catcagcctg gaccgctacc tgctcatcct ctcgccgctg
1081 cgctacaagc tgcgcatgac agccccgcgt gccctggcgc tcatcctggg tgcctggagc
1141 ctcgccgcgc tggcctcctt cctgcccctc ttgctgggct ggcatgaact gggcaaagct
1201 cgaacatctg ccccgggcca gtgccgccta ttggccagcc tgccttatgt cctcgtggcg
1261 tccggcgtca cctttttcct gccttcgggt gccatctgct tcacctactg caggatcttg
1321 ctggctgccc gcaagcaggc ggtgcaagtg gcctcgctca ccacgggcac ggctacggct
1381 ggccaggcct tggaaacctt gcaggtgccc aggacaccac gcccagggat ggagtctgct
1441 gacagtaggc gtctgaccac caagcatagc aggaaggcct tgaaggccag cctgaccctc
1501 ggcatcctgc tgagcatgtt ctttgtcacc tggctgccct tctttgtggc cagcatagct
1561 caggccgtat gtgactgcat ctctccaggt ctcttcgatg tcctcacatg gctgggatac
1621 tgtaatagca ccatgaaccc tatcatctat cccctcttca tgcgggactt caagagggcc
1681 ctgggcaggt tcgtgccgtg tgtccactgt cccccggagc accgggccag ccccgcctcc
1741 ccctccatgt ggacctctca cagtggtgcc aggccaggcc tcagcctgca gcaggtgctg
1801 cccctgcctc tgccacccaa ctcagattca gactcagctt cagggggcac ctcgggcctg
1861 cagctcacag cccagctttt gctgcctgga gaggcgaccc gggacccccc gccacccacc
1921 agggccccta ctgtggtcaa cttcttcgtc acagactctg tggagcctga gatacggcag
1981 catccacttg gttcccccat gaacagtgct ggtggtagcg caggaggagg atccaccatg
2041 gtcgactcat cacgtcgtaa gtggaataag acaggtcacg cagtcagagc tataggtcgg
2101 ctgagctcac tcgagaacgt ctatatcaag gccgacaagc agaagaacgg catcaaggcg
2161 aacttcaaga tccgccacaa catcgaggac ggcggcgtgc agctcgccta ccactaccag
2221 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcgtg
2281 cagtccatac tttcgaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
2341 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggcgg taccggaggg
2401 agcatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcca ggtcgagctg
2461 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggtgaggg cgatgccacc
2521 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
2581 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
2641 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acatccagga gcgcaccatc
2701 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
2761 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
2821 cacaagctgg agtacaacac gcgtgaccaa ctgactgaag agcagatcgc agaatttaaa
2881 gaggctttct ccctatttga caaggacggg gatgggacga taacaaccaa ggagctgggg
2941 acggtgatgc ggtctctggg gcagaacccc acagaagcag agctgcagga catgatcaat
3001 gaagtagatg ccgacggtga cggcacaatc gacttccctg agttcctgac aatgatggca
3061 agaaaaatga aagacacaga cagtgaagaa gaaattagag aagcgttccg tgtgtttgat
3121 aaggacggca atggctacat cggcgcagca gagcttcgcc acgtgatgac aaaccttgga
3181 gagaagttaa cagatgaaga ggttgatgaa atgatcaggg tagcagacat cgatggggat
3241 ggtcaggtaa actacgaaga gtttgtacaa atgatgacag cgaagtaaga attctgcaga
3301 tatccatcac actggcggcc gctcgagcat gcatctagag ggccctattc tatagtgtca
3361 cctaaatgct agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
3421 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
3481 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
3541 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
3601 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gtccggaact
3661 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
3721 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
3781 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
3841 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
3901 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
3961 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
4021 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
4081 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
4141 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
4201 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
4261 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
4321 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
4381 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
4441 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
4501 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
4561 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
4621 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
4681 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
4741 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
4801 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
4861 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
4921 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
4981 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
5041 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
5101 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
5161 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
5221 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
5281 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
5341 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
5401 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
5461 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
5521 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
5581 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
5641 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
5701 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
5761 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
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