| 质粒名称: | pcDNA3 hML23Cm6 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 109368 |
| 存储实验室: | Robert Lucas |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5366 bp |
| 完整质粒大小: | 6410 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: | human Melanopsin-mGluR6 chimera,hML23Cm6 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1119 bp |
| GenBank编号 | NM_033282, NM_000843.3 |
| 启动子 | CMV |
| 标签/融合蛋白: | 1D4（插入件上的C端子） |
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | GGAGGTCTATATAAGCAGAGC |
| 3' 测序引物 | ggcaccttccagggtcaagg |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3 hML23Cm6 was a gift from Robert Lucas (Addgene plasmid # 109368 ; http://n2t.net/addgene:109368 ; RRID:Addgene_109368) |
| 参考文献部分: | A live cell assay of GPCR coupling allows identification of optogenetic tools for controlling Go and Gi signaling. Ballister ER, Rodgers J, Martial F, Lucas RJ. BMC Biol. 2018 Jan 16;16(1):10. doi: 10.1186/s12915-017-0475-2. 10.1186/s12915-017-0475-2 [pii] PubMed 29338718 |
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 pcDNA3 hML23Cm6 6488 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Human melanopsin chimera with intracellular loops 2-3 and C-terminus
from mGluR6.
ACCESSION .
VERSION .
KEYWORDS pcDNA3 hML23Cm6
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6488)
AUTHORS Ballister ER, Rodgers J, Martial F, Lucas RJ
TITLE A live cell assay of GPCR coupling allows identification of
optogenetic tools for controlling Go and Gi signaling.
JOURNAL BMC Biol. 2018 Jan 16;16(1):10. doi: 10.1186/s12915-017-0475-2.
PUBMED 29338718
REFERENCE 2 (bases 1 to 6488)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6488
/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"
primer_bind complement(1845..1862)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1851..2075
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2121..2549
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2208..2227)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2418..2439
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(2558..2578)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2563..2892
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2743..2878
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 2805..2824
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 2959..3753
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3013..3032)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 3623..3642
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 3927..4048
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3964..3983)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4018..4037
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4097..4113)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4097..4113)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4110..4132)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4121..4137
/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(4145..4175)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4190..4211
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4328..4345)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4499..5087)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4579..4598)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5258..6118)
/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 5881..5900
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6119..6223)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6298..6317)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gtcattgatt 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 acccaagctg
661 gctagcgttt aaacttaagc ttgccaccat gaaccctcct tcggggccaa gagtcccgcc
721 cagcccaacc caagagccca gctgcatggc caccccagca ccacccagct ggtgggacag
781 ctcccagagc agcatctcca gcctgggccg gcttccatcc atcagtccca cagcacctgg
841 gacttgggct gctgcctggg tccccctccc cacggttgat gttccagacc atgcccacta
901 taccctgggc acagtgatct tgctggtggg actcacgggg atgctgggca acctgacggt
961 catctatacc ttttgcagga gcagaagcct ccggacacct gccaacatgt tcattatcaa
1021 cctcgcggtc agcgacttcc tcatgtcctt cacccaggcc cctgtcttct tcaccagtag
1081 cctctataag cagtggctct ttggggagac aggctgcgag ttctatgcct tctgtggagc
1141 tctctttggc atttcctcca tgatcaccct gacggccatc gccctggacc gtatctaccg
1201 catctttgag cagggcaagc gctcggtcac accccctccc ttcatcagcc ccacctcaca
1261 ggtcctgctg ggcgtttggc tctatgccct ggcctggagt ctgccaccct tcttcggctg
1321 gagcgcctac gtgcccgagg ggttgctgac atcctgctcc tgggactaca tgagcttcac
1381 gccggccgtg cgtgcctaca ccatgcttct ctgctgcttc gtgttcttcc tccctctgct
1441 tatcatcatc tactgctaca tcttcatctt cagggcccgt ggcgtgcccg agaccttcaa
1501 cgaggccaag atcatgctgc tggtcatcct cctcttcgtg ctctcctggg ctccctattc
1561 cgctgtggcc ctggtggcct ttgctgggta cgcacacgtc ctgacaccct acatgagctc
1621 ggtgccagcc gtcatcgcca aggcctctgc aatccacaac cccatcattt acgccatcac
1681 ccaccccgag cagaatgtgc agaagcgaaa gcggagcctc aaggccacct ccacggtggc
1741 agccccaccc aagggcgagg atgcagaggc ccacaagacg gagactagcc aggtggcccc
1801 ggcctagctc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
1861 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
1921 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
1981 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
2041 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
2101 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
2161 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
2221 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
2281 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
2341 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
2401 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
2461 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
2521 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
2581 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
2641 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
2701 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
2761 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
2821 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
2881 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
2941 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
3001 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
3061 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
3121 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
3181 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
3241 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
3301 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
3361 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
3421 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
3481 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
3541 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
3601 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
3661 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
3721 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
3781 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
3841 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
3901 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
3961 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
4021 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
4081 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
4141 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
4201 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
4261 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
4321 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
4381 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
4441 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
4501 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
4561 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
4621 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
4681 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
4741 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
4801 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
4861 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
4921 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
4981 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
5041 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
5101 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
5161 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
5221 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
5281 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
5341 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
5401 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
5461 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
5521 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
5581 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
5641 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
5701 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
5761 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
5821 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
5881 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
5941 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
6001 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
6061 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
6121 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
6181 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
6241 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
6301 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
6361 cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg
6421 catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata
6481 tacgcgtt
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
- 上一篇：lenti_dCas9-KRAB-MeCP2
- 下一篇：eGFPC1 FKBP
