| 质粒名称: | pcDNA-4mtD3cpv |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 36324 |
| 存储实验室: | Amy Palmer, Roger Tsien |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3 |
| 载体出品公司: | Invitrogen |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5400 bp |
| 完整质粒大小: | 7800 bp |
| 载体修饰: | 线粒体靶向序列N-末端插入 |
| 载体类型: | -- |
| 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: | 4mtD3cpv second generation cameleon (calcium sensor) targeted to mitochondria,D family cameleon |
| 物种来源: | Synthetic |
| 插入大小: | 2370 bp |
| 基因突变: | -- |
| GenBank编号 | DQ479429.1 |
| 启动子 | CMV |
| 标签/融合蛋白: | 人COX亚单位8a的前29个氨基酸（主干上的N末端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI（未销毁） |
| 3' 克隆位点: | EcoRI（未销毁） |
| 5' 测序引物 | T7 Forward |
| 3' 测序引物 | BGH Reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA-4mtD3cpv was a gift from Amy Palmer & Roger Tsien (Addgene plasmid # 36324 ; http://n2t.net/addgene:36324 ; RRID:Addgene_36324) |
| 参考文献部分: | Ca2+ indicators based on computationally redesigned calmodulin-peptide pairs. Palmer AE, Giacomello M, Kortemme T, Hires SA, Lev-Ram V, Baker D, Tsien RY. Chem Biol. 2006 May;13(5):521-30. 10.1016/j.chembiol.2006.03.007 PubMed 16720273 |
买家导航
pcDNA4mtD3cpv载体基本信息
pcDNA4mtD3cpv载体图谱质粒图谱和多克隆位点信息
pcDNA4mtD3cpv载体简介
pcDNA4mtD3cpv载体序列
LOCUS pcDNA-4mtD3cpv 7754 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pcDNA-4mtD3cpv
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7754)
AUTHORS Palmer AE, Giacomello M, Kortemme T, Hires SA, Lev-Ram V, Baker D,
Tsien RY
TITLE Ca2+ indicators based on computationally redesigned
calmodulin-peptide pairs.
JOURNAL Chem Biol. 2006 May;13(5):521-30.
PUBMED 16720273
REFERENCE 2 (bases 1 to 7754)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7754
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(83..943)
/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 706..725
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(944..1048)
/gene="bla"
/label=AmpR promoter
primer_bind complement(1123..1142)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 1314..1693
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1694..1897
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1848..1868
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 1942..1961
/label=T7
/note="T7 promoter, forward primer"
promoter 1942..1960
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
regulatory 2358..2367
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 3804..4322
/codon_start=1
/product="N-terminal fragment of mVenus for use in
bimolecular fluorescence complementation (BiFC) (Kodama and
Hu, 2010)"
/label=VN173
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
ANFKIRHNIE"
promoter complement(4386..4404)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(4387..4404)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(4424..4441)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 4430..4654
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 4700..5128
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4787..4806)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4997..5018
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(5137..5157)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 5142..5471
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 5322..5457
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 5384..5403
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5538..6332
/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(5592..5611)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6202..6221
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 6506..6627
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6543..6562)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6597..6616
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6676..6692)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6676..6692)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6689..6711)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6700..6716
/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(6724..6754)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6769..6790
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6907..6924)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7078..7666)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7158..7177)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
61 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
121 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
181 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
241 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
301 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
361 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
421 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
481 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
541 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
601 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
661 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
721 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
781 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
841 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
901 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
961 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
1021 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
1081 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
1141 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
1201 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
1261 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
1321 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
1381 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
1441 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
1501 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
1561 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
1621 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
1681 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
1741 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
1801 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
1861 taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac
1921 tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagacccaag cttatgtccg
1981 tcctgacgcc gctgctgctg cggggcttga caggctcggc ccggcggctc ccagtgccgc
2041 gcgccaagat ccattcgttg ggggatccca tgtccgtcct gacgccgctg ctgctgcggg
2101 gcttgacagg ctcggcccgg cggctcccag tgccgcgcgc caagatccat tcgttgggga
2161 agcttatgtc cgtcctgacg ccgctgctgc tgcggggctt gacaggctcg gcccggcggc
2221 tcccagtgcc gcgcgccaag atccattcgt tgggggatcc catgtccgtc ctgacgccgc
2281 tgctgctgcg gggcttgaca ggctcggccc ggcggctccc agtgccgcgc gccaagatcc
2341 attcgttggg gaagcttgcc accatggtga gcaagggcga ggagctgttc accggggtgg
2401 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caggttcagc gtgtccggcg
2461 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
2521 agctgcccgt gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttca
2581 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
2641 acgtccagga gcgtaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
2701 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
2761 aggacggcaa catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata
2821 tcaccgccga caagcagaag aacggcatca aggcccactt caagatccgc cacaacatcg
2881 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
2941 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
3001 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgcccgc atgcatgacc
3061 aactgacaga agagcagatt gcagagttca aagaagcctt ctcattattg gacaaggatg
3121 gggacggcac catcaccaca aaggaacttg gcaccgctct gaggtcgctt ggacaaaacc
3181 caacggaagc agaattgcag gatatgatca atgaagtcga tgctgatggc aatggaacga
3241 tttactttcc tgaatttctt actatgatgg ctagaaaaat gaaggacaca gacagcgaag
3301 aggaaatccg agaagcattc cgtgtttttg acaaggatgg gaacggctac atcagcgctg
3361 ctgaattacg tcacgtcatg acaaacctcg gggagaagtt aacagatgaa gaagttgatg
3421 aaatgataag ggaagcagat atcgatggtg atggccaagt aaactatgaa gagtttgtac
3481 aaatgatgac agcaaagggg gggaagaggc gctggcagaa aacaggccat gctgtccgag
3541 cattcggaag actgaagaag atctccagct ccggggcact ggagctcatg gacggcggcg
3601 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
3661 ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg
3721 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
3781 tgtacaaggg tggcagcggt ggcatggtga gcaagggcga ggagctgttc accggggtgg
3841 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
3901 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca
3961 agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctg cagtgcttcg
4021 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
4081 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
4141 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
4201 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
4261 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg
4321 agtaagaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagggc
4381 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
4441 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
4501 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
4561 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
4621 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
4681 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
4741 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
4801 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
4861 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
4921 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
4981 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
5041 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
5101 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
5161 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
5221 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
5281 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
5341 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
5401 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
5461 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
5521 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
5581 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
5641 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
5701 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
5761 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
5821 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
5881 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
5941 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
6001 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
6061 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
6121 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
6181 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
6241 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
6301 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
6361 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
6421 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
6481 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
6541 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
6601 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
6661 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
6721 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
6781 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
6841 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
6901 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
6961 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
7021 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
7081 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
7141 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
7201 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
7261 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
7321 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
7381 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
7441 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
7501 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
7561 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
7621 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
7681 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
7741 gagattatca aaaa
//
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