| 质粒名称: | pMXs-3XHA-EGFP-OMP25 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 83356 |
| 存储实验室: | David Sabatini |
|
载体骨架基本信息 |
|
| 载体名称: | pMXs rtv-016 |
| 载体出品公司: | Cell BioLabs |
| 空载体大小: | 5600 bp |
| 完整质粒大小: | 6600 bp |
| 载体修饰: | 原始载体rtv-016的所有限制性位点均完整：克隆插入带有XhoI和NotI的MCS |
| 载体类型: | 哺乳动物表达，逆转录病毒 |
| 筛选标记: | 杀稻瘟菌素 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 30 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | 3XHA-EGFP-OMP25 |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pMXs-3XHA-EGFP-OMP25 was a gift from David Sabatini (Addgene plasmid # 83356 ; http://n2t.net/addgene:83356 ; RRID:Addgene_83356) |
| 参考文献部分: | Absolute Quantification of Matrix Metabolites Reveals the Dynamics of Mitochondrial Metabolism. Chen WW, Freinkman E, Wang T, Birsoy K, Sabatini DM. Cell. 2016 Aug 25;166(5):1324-1337.e11. doi: 10.1016/j.cell.2016.07.040. 10.1016/j.cell.2016.07.040 PubMed 27565352 |
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 pMXs-3XHA-EGFP-OMP25 6597 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION For tagging mitochondria with HA epitopes (HA-MITO construct).
ACCESSION .
VERSION .
KEYWORDS pMXs-3XHA-EGFP-OMP25
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6597)
AUTHORS Chen WW, Freinkman E, Wang T, Birsoy K, Sabatini DM
TITLE Absolute Quantification of Matrix Metabolites Reveals the Dynamics
of Mitochondrial Metabolism.
JOURNAL Cell. 2016 Aug 25;166(5):1324-1337.e11. doi:
10.1016/j.cell.2016.07.040.
PUBMED 27565352
REFERENCE 2 (bases 1 to 6597)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6597
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 22..615
/note="long terminal repeat from Moloney murine leukemia
virus"
misc_feature 678..1035
/label=MMLV Psi
/note="packaging signal of Moloney murine leukemia virus
(MMLV)"
CDS 1094..1510
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1440..1462
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1478..1494
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
misc_feature 1520..1894
/label=pol region
/note="Moloney murine leukemia virus (MMLV) pol region
containing the splice acceptor site"
primer_bind 1832..1848
/label=pBMN 5'
/note="MMLV sequence, for inserts in pBMN retroviral
vector"
CDS 1968..1994
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 2007..2033
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 2046..2072
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 2085..2798
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
CDS 2085..2798
/codon_start=1
/product="the original enhanced GFP (Yang et al., 1996)"
/label=EGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind complement(2127..2148)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(2388..2407)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 2735..2756
/label=EGFP-C
/note="EGFP, forward primer"
misc_feature 2949..3535
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
primer_bind complement(3116..3133)
/label=IRES reverse
/note="IRES internal ribosome entry site, reverse primer.
Also called pCDH-rev"
primer_bind 3343..3362
/label=IRES-F
/note="IRES internal ribosome entry site, forward primer"
LTR 4102..4571
/note="long terminal repeat from Moloney murine leukemia
virus"
primer_bind complement(4701..4718)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4872..5460)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4952..4971)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5624..6484)
/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 6247..6266
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6485..6589)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat
1921 cccagtgtgg tggtacggga attcctgcag gcctcgaggc caccatgtat ccctatgacg
1981 tgcctgatta cgccggcaca ggatcctacc cctatgatgt gcctgactac gctggcagcg
2041 ccggataccc ttatgatgtg cctgattatg ctggagggag cggcgtgagc aagggcgagg
2101 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
2161 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
2221 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
2281 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
2341 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
2401 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
2461 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
2521 acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
2581 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
2641 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg
2701 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
2761 ccgccgggat cactctcggc atggacgagc tgtacaagcc gcggcatcga ggcgacggag
2821 agcctagtgg agttcctgta gctgtggtgc tgctgccagt gtttgccctt accctggtag
2881 cagtttgggc cttcgtgaga taccgaaagc agctctgagc ggccgctacg taaattccgc
2941 cccccccccc cctctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
3001 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
3061 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
3121 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
3181 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
3241 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
3301 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
3361 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
3421 cctcggtgca catgctttac atgtgtttag tcgaggttaa aaaaacgtct aggccccccg
3481 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggcca caaccatggt
3541 tcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat
3601 ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt
3661 cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg
3721 cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa
3781 caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg
3841 gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt
3901 gctgccctct ggttatgtgt gggagggcta agtcgacgat aaaataaaag attttattta
3961 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
4021 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa
4081 ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
4141 cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat
4201 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
4261 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
4321 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
4381 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc
4441 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
4501 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
4561 ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt aagtatttac
4621 attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
4681 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
4741 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
4801 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
4861 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
4921 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
4981 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
5041 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
5101 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
5161 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
5221 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
5281 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
5341 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
5401 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
5461 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
5521 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gcggccggcc
5581 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
5641 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
5701 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
5761 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
5821 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
5881 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
5941 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
6001 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
6061 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
6121 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
6181 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
6241 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
6301 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
6361 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
6421 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
6481 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
6541 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttc
//
