| 质粒名称: | pHAGE-mt-mKeima |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 131626 |
| 存储实验室: | Richard Youle |
|
载体骨架基本信息 |
|
| 载体名称: | pHAGE |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6436 bp |
| 完整质粒大小: | 7222 bp |
| 载体类型: | 哺乳细胞表达,慢病毒 |
| 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: | mt-mKeima |
| 物种来源: | stony coral Montipora |
| 插入大小: | 786 bp |
| 启动子 | CMV |
| 克隆方法: | Gibson 克隆 |
| 5' 测序引物 | CGCAAATGGGCGGTAGGCGTG |
| 3' 测序引物 | ATAGCGTAAAAGGAGCAACA |
|
文献引用方法 |
|
| 材料和方法部分: | pHAGE-mt-mKeima was a gift from Richard Youle (Addgene plasmid # 131626 ; http://n2t.net/addgene:131626 ; RRID:Addgene_131626) |
| 参考文献部分: | Spatiotemporal Control of ULK1 Activation by NDP52 and TBK1 during Selective Autophagy. Vargas JNS, Wang C, Bunker E, Hao L, Maric D, Schiavo G, Randow F, Youle RJ. Mol Cell. 2019 Apr 18;74(2):347-362.e6. doi: 10.1016/j.molcel.2019.02.010. Epub 2019 Mar 7. 10.1016/j.molcel.2019.02.010 PubMed 30853401 |
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 pHAGE-mt-mKeima 7222 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION lentiviral expression of mitochondrial mKeima for generation of
stable cell lines.
ACCESSION .
VERSION .
KEYWORDS pHAGE-mt-mKeima
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7222)
AUTHORS Vargas JNS, Wang C, Bunker E, Hao L, Maric D, Schiavo G, Randow F,
Youle RJ
TITLE Spatiotemporal Control of ULK1 Activation by NDP52 and TBK1 during
Selective Autophagy.
JOURNAL Mol Cell. 2019 Apr 18;74(2):347-362.e6. doi:
10.1016/j.molcel.2019.02.010. Epub 2019 Mar 7.
PUBMED 30853401
REFERENCE 2 (bases 1 to 7222)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7222
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 71..188
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
enhancer 240..543
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 544..747
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 698..718
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 744..768
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 828..902
/codon_start=1
/product="mitochondrial presequence of human cytochrome c
oxidase subunit VIII (Rizutto et al., 1989)"
/label=COX8 presequence
/translation="MSVLTPLLLRGLTGSARRLPVPRAK"
CDS 945..1610
/codon_start=1
/product="human codon-optimized monomeric red fluorescent
protein with a large Stokes shift (Guan et al. 2015)"
/label=hmKeima8.5
/translation="MVSVIAKQMTYKVYMSGTVNGHYFEVEGDGKGKPYEGEQTVKLTV
TKGGPLPFAWDILSPQLQYGSIPFTKYPEDIPDYFKQSFPEGYTWERSMNFEDGAVCTV
SNDSSIQGNCFIYNVKISGENFPPNGPVMQKKTQGWEPSTERLFARDGMLIGNDYMALK
LEGGGHYLCEFKSTYKAKKPVRMPGRHEIDRKLDVTSHNRDYTSVEQCEIAIARHSLLG
"
misc_feature 1621..2209
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(1674..1694)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2092..2103)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 2284..2517
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(2636..2654)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 2722..2826
/gene="bla"
/label=AmpR promoter
CDS 2827..3687
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(3045..3064)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 3858..4446
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4347..4366
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4600..4617
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 4709..4728
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
protein_bind 4857..4878
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 4893..4923
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4931..4947
/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)."
primer_bind 4936..4958
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 4955..4971
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 4955..4971
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 5021..5040
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
LTR 5232..5865
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 5912..6037
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6534..6767
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 6952..6996
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
ORIGIN
1 ccattcgatt agtgaacgga tctcgacggt atcgccgaat taattcacaa atggcagtat
61 tcatccacaa ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
121 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc
181 aaaattttcg ggtttattac agggacagca gagatccagt ttggactagt ggagttccgc
241 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
301 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
361 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
421 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
481 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
541 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
601 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
661 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
721 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
781 catccacgct gttttgacct ccatagaaga caccggcggc cgccaccatg tccgtcctga
841 cgccgctgct gctgcggggc ttgacaggct cggcccggcg gctcccagtg ccgcgcgcca
901 agatccattc gttgccgccg gaggggaagc tcgggggatc cgccatggtg agcgtgatcg
961 ccaagcagat gacctacaag gtgtacatga gcggcaccgt gaacggccac tacttcgagg
1021 tggagggcga cggcaagggc aagccctacg agggcgagca gaccgtgaag ctgaccgtga
1081 ccaagggtgg ccccctgccc ttcgcctggg acatcctgag cccccagctc cagtacggca
1141 gcatcccctt caccaagtac cccgaggaca tccccgacta cttcaagcag agcttccccg
1201 agggctacac ctgggagcgc agcatgaact tcgaggacgg cgccgtgtgc accgtgagca
1261 acgacagcag catccagggc aactgcttca tctacaacgt gaagatcagc ggcgagaact
1321 tcccccccaa cggccccgtg atgcagaaga agacccaggg ctgggagccc agcaccgagc
1381 gcctgttcgc ccgcgacgga atgctgatcg gcaacgacta catggccctg aagctggagg
1441 gcggcggcca ctacctgtgc gagttcaaga gcacctacaa ggccaagaag cccgtgagga
1501 tgcccggccg ccacgagatc gaccgcaagc tggacgtgac cagccacaac cgcgactaca
1561 ccagcgtgga gcagtgcgag atcgccatcg cccgccactc cctgctgggc taaggatcct
1621 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
1681 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
1741 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
1801 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
1861 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
1921 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
1981 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
2041 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
2101 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
2161 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgcc tgagatcctt
2221 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
2281 gactggaagg gctaattcac tcccaacgaa gacaagatct gctttttgct tgtactgggt
2341 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
2401 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
2461 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagta
2521 gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga atatcagaga
2581 gtgagaggcc cgggttaatt aaggaaaggg ctagatcatt cttgaagacg aaagggcctc
2641 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
2701 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
2761 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
2821 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
2881 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
2941 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
3001 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
3061 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
3121 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
3181 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
3241 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
3301 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
3361 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
3421 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
3481 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
3541 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
3601 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
3661 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
3721 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
3781 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
3841 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
3901 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
3961 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
4021 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
4081 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
4141 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
4201 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
4261 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
4321 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
4381 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
4441 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
4501 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
4561 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
4621 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
4681 agcaagctca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
4741 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
4801 tccccgtggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
4861 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
4921 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg
4981 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
5041 aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa
5101 cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc
5161 tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca
5221 aacttagtag ttggaagggc taattcactc ccaaagaaga caagatatcc ttgatctgtg
5281 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt
5341 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt
5401 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat
5461 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca
5521 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa
5581 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg
5641 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag
5701 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
5761 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
5821 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag
5881 ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga
5941 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
6001 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
6061 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
6121 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
6181 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
6241 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
6301 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
6361 caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga tatgagggac
6421 aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt aggagtagca
6481 cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg aataggagct
6541 ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc aatgacgctg
6601 acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa tttgctgagg
6661 gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa gcagctccag
6721 gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg gatttggggt
6781 tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg gagtaataaa
6841 tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga aattaacaat
6901 tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga aaagaatgaa
6961 caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa cataacaaat
7021 tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg tttaagaata
7081 gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc attatcgttt
7141 cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga agaagaaggt
7201 ggagagagag acagagacag at
//
