| 质粒名称: | pDEST47-MCU-GFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 31732 |
| 存储实验室: | Vamsi Mootha |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA6.2/C-EmGFP-DEST |
| 载体出品公司: | Invitrogen (Cat#V355-20) |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5784 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 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: | MCU,CCDC109A |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1056 bp |
| 基因突变: | -- |
| GenBank编号 | NM_138357 |
| 启动子 | CMV |
| 标签/融合蛋白: | GFP（主干上的C终端） |
| 克隆方法: | Gateway克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | T7 |
| 3' 测序引物 | GAACTTCAGGGTCAGCTTGC |
|
文献引用方法 |
|
| 材料和方法部分: | pDEST47-MCU-GFP was a gift from Vamsi Mootha (Addgene plasmid # 31732 ; http://n2t.net/addgene:31732 ; RRID:Addgene_31732) |
| 参考文献部分: | Integrative genomics identifies MCU as an essential component of the mitochondrial calcium uniporter. Baughman JM, Perocchi F, Girgis HS, Plovanich M, Belcher-Timme CA, Sancak Y, Bao XR, Strittmatter L, Goldberger O, Bogorad RL, Koteliansky V, Mootha VK. Nature. 2011 Jun 19. ():. 10.1038/nature10234 PubMed 21685886 |
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 pDEST47-MCU-GFP 6840 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pDEST47-MCU-GFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6840)
AUTHORS Baughman JM, Perocchi F, Girgis HS, Plovanich M, Belcher-Timme CA,
Sancak Y, Bao XR, Strittmatter L, Goldberger O, Bogorad RL,
Koteliansky V, Mootha VK
TITLE Integrative genomics identifies MCU as an essential component of the
mitochondrial calcium uniporter.
JOURNAL Nature. 2011 Jun 19. ():.
PUBMED 21685886
REFERENCE 2 (bases 1 to 6840)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6840
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 105..484
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 485..688
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 639..659
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 733..752
/label=T7
/note="T7 promoter, forward primer"
promoter 733..751
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 785..809
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
protein_bind complement(1863..1887)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 1901..2620
/codon_start=1
/product="Emerald GFP"
/label=EmGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(1946..1967)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2207..2226)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2554..2575
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(2648..2667)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 2692..2740
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 2942..3370
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3029..3048)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3239..3260
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3379..3399)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3384..3713
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3564..3699
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3626..3645
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 3761..3808
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 3827..4225
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
polyA_signal 4383..4504
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4420..4439)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4474..4493
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4553..4569)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4553..4569)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4566..4588)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4577..4593
/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(4601..4631)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4646..4667
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4784..4801)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4955..5543)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5035..5054)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5714..6574)
/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 6337..6356
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6575..6679)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6754..6773)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
61 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
121 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
181 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
241 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
301 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
361 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
421 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
481 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
541 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
601 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
661 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
721 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt taagctgagc
781 atcaacaagt ttgtacaaaa aagttggcat ggcggccgcc gcaggtagat cgctcctgct
841 gctcctctcc tctcggggcg gcggcggcgg gggcgccggc ggctgcgggg cgctgactgc
901 cggctgcttc cctgggctgg gcgtcagccg ccaccggcag cagcagcacc accggacggt
961 acaccagagg atcgcttcct ggcagaattt gggagctgtt tattgcagca ctgttgtgcc
1021 ctctgatgat gttacagtgg tttatcaaaa tgggttacct gtgatatctg tgaggctacc
1081 atcccggcgt gaacgctgtc agttcacact caagcctatc tctgactctg ttggtgtatt
1141 tttacgacaa ctgcaagaag aggatcgggg aattgacaga gttgctatct attcaccaga
1201 tggtgttcgc gttgctgctt caacaggaat agacctcctc ctccttgatg actttaagct
1261 ggtcattaat gacttaacat accacgtacg accaccaaaa agagacctct taagtcatga
1321 aaatgcagca acgctgaatg atgtaaagac attggtccag caactataca ccacactgtg
1381 cattgagcag caccagttaa acaaggaaag ggagcttatt gaaagactag aggatctcaa
1441 agagcagctg gctcccctgg aaaaggtacg aattgagatt agcagaaaag ctgagaagag
1501 gaccactttg gtgctatggg gtggccttgc ctacatggcc acacagtttg gcattttggc
1561 ccggcttacc tggtgggaat attcctggga catcatggag ccagtaacat acttcatcac
1621 ttatggaagt gccatggcaa tgtatgcata ttttgtaatg acacgccagg aatatgttta
1681 tccagaagcc agagacagac aatacttact atttttccat aaaggagcca aaaagtcacg
1741 ttttgaccta gagaaataca atcaactcaa ggatgcaatt gctcaggcag aaatggacct
1801 taagagactg agagacccat tacaagtaca tctgcctctc cgacaaattg gtgaaaaaga
1861 ttgcccaact ttcttgtaca aagtggttga tgctgttaac atggtgagca agggcgagga
1921 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
1981 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
2041 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcaccta
2101 cggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
2161 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
2221 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
2281 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
2341 cagccacaag gtctatatca ccgccgacaa gcagaagaac ggcatcaagg tgaacttcaa
2401 gacccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
2461 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
2521 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
2581 cgccgggatc actctcggca tggacgagct gtacaagtaa tgataagttt aaacggggga
2641 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
2701 aaagacagaa taaaacgcac gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
2761 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
2821 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
2881 gtcggggcgg caggccctgc catagcagat ctgcgcagct ggggctctag ggggtatccc
2941 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
3001 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
3061 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
3121 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
3181 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
3241 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
3301 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
3361 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
3421 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
3481 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
3541 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
3601 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
3661 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
3721 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
3781 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatgg ccaagccttt
3841 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
3901 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
3961 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
4021 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
4081 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
4141 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
4201 ctctggttat gtgtgggagg gctaagcact tcgtggccga ggagcaggac tgacacgtgc
4261 tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
4321 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
4381 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
4441 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
4501 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
4561 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
4621 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
4681 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
4741 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
4801 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
4861 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
4921 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
4981 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
5041 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
5101 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
5161 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
5221 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
5281 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
5341 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
5401 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
5461 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
5521 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
5581 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
5641 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
5701 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
5761 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
5821 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
5881 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
5941 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
6001 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
6061 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
6121 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
6181 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
6241 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
6301 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
6361 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
6421 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
6481 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
6541 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
6601 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
6661 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
6721 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
6781 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
//
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