| 质粒名称: | GPD-roGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 49436 |
| 存储实验室: | Paul Schumacker |
|
载体骨架基本信息 |
|
| 载体名称: | VQ Ad5CMV K-NpA |
| 载体出品公司: | ViraQuest Inc. |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5217 bp |
| 完整质粒大小: | 6200 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 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: | GPD-roGFP2 |
| 物种来源: | -- |
| 插入大小: | 3150 bp |
| 基因突变: | -- |
| GenBank编号 | JN247409 |
| 启动子 | CMV |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | GPD-roGFP was a gift from Paul Schumacker (Addgene plasmid # 49436 ; http://n2t.net/addgene:49436 ; RRID:Addgene_49436) |
| 参考文献部分: | Hypoxia triggers subcellular compartmental redox signaling in vascular smooth muscle cells. Waypa GB, Marks JD, Guzy R, Mungai PT, Schriewer J, Dokic D, Schumacker PT. Circ Res. 2010 Feb 19;106(3):526-35. doi: 10.1161/CIRCRESAHA.109.206334. Epub 2009 Dec 17. 10.1161/CIRCRESAHA.109.206334 PubMed 20019331 |
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 GPD-roGFP 8598 bp ds-DNA circular SYN 30-JUL-2019
DEFINITION Expresses thiol redox-sensitive ratiometric sensor roGFP in the
mitochondrial intermembrane space of mammalian cells.
ACCESSION .
VERSION .
KEYWORDS GPD-roGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8598)
AUTHORS Waypa GB, Marks JD, Guzy R, Mungai PT, Schriewer J, Dokic D,
Schumacker PT
TITLE Hypoxia triggers subcellular compartmental redox signaling in
vascular smooth muscle cells.
JOURNAL Circ Res. 2010 Feb 19;106(3):526-35. doi:
10.1161/CIRCRESAHA.109.206334. Epub 2009 Dec 17.
PUBMED 20019331
REFERENCE 2 (bases 1 to 8598)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jul 30, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8598
/organism="synthetic DNA construct"
/mol_type="other DNA"
repeat_region 1..103
/label=ITR
/note="inverted terminal repeat of human adenovirus
serotype 5"
misc_signal 191..341
/label=Ad5 Psi
/note="packaging signal for adenovirus serotype 5"
enhancer 370..673
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 674..877
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 828..848
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 1339..1348
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2900..3619
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(2945..2966)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(3206..3225)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 3553..3574
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(3664..3686)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin complement(6761..7349)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6841..6860)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7520..8380)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 8143..8162
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8381..8485)
/gene="bla"
/label=AmpR promoter
primer_bind 8553..8571
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 catcatcaat aatatacctt attttggatt gaagccaata tgataatgag ggggtggagt
61 ttgtgacgtg gcgcggggcg tgggaacggg gcgggtgacg tagtagtgtg gcggaagtgt
121 gatgttgcaa gtgtggcgga acacatgtaa gcgacggatg tggcaaaagt gacgtttttg
181 gtgtgcgccg gtgtacacag gaagtgacaa ttttcgcgcg gttttaggcg gatgttgtag
241 taaatttggg cgtaaccgag taagatttgg ccattttcgc gggaaaactg aataagagga
301 agtgaaatct gaataatttt gtgttactca tagcgcgtaa tatttgtcta gggagatcag
361 cctgcaggtc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
421 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
481 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
541 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
601 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
661 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
721 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
781 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
841 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatggt
901 accgtttaaa ctcgagcggc cgccagtgtg atggatatct gcagaattcc gctcgccgac
961 ccctggcgtg tgacgaccga tctcaggctg agcaatggcg tttcaaaagg tagtgaaagg
1021 gactattctt atgggtggag gagctctggc cactgttttg ggactctctc agtttgctca
1081 ctacagaagg aagcaagtaa gcctggcgta tgtggaagca gcaacatgct tctcagagcc
1141 cgtgaacagg gagcctcctt ccagagaagc tcagctcatg actctgcaga acacatcgga
1201 atttgacatc ctcgtcattg gaggaggagc cacaggatgt ggctgtgcgc tagacgccgt
1261 caccagagga ctgaaaacag cccttgtaga gagaaatgac ttcgcatcag ggactagcag
1321 cagaagcact aaattgatcc atggtggcgt acgatacctc cagaaggcta tcacgaactt
1381 ggatgttgaa cagtatagga tggtgaaaga ggcccttcat gaacgtgcca acttactaga
1441 aatcgctccc catctatcag ctccgctgcc tataatgctg ccactttata agtggtggca
1501 gttaccttat tactgggtgg gaatcaagat gtatgatctg gttgcaggaa gtcattgcct
1561 gaagagcagt tatgtcctca gtaaatccag agcccttgag catttcccca tgctccagaa
1621 ggacaagctg gtcggagcca ttgtctacta tgatggacag cacaatgacg cccggatgaa
1681 cctcgccatc gccctcaccg ctgccaggta cggggctgcc acggccaatt acatggaggt
1741 ggtgagcttg ctcaagaaga cagaccccga aactgggaaa gagcgcgtga gtggtgcccg
1801 gtgcaaggac gtgctcacag ggcatgagtt taacgtgaga gctaaatgcg ttatcaatgc
1861 caccggccct ttcacagact ccgtgcgcaa aatggatgat aacgatgttg ttcccatctg
1921 ccagcccagt gcaggggttc acattgtgat gcccgggtac tacagccccg agaacatggg
1981 acttcttgat cctgcaacca gtgatggcag agttattttc ttcttgcctt gggagaagat
2041 gacaattgcg ggtaccactg attcgccaac ggacgtcacg caccatccta ttccttcgga
2101 agacgacatt aacttcatct taaatgaagt gcggaactac ctgagttgcg acgttgaagt
2161 gagaagaggg gatgtcctgg cagcgtggag tggcattcgc ccgcttgtta ccgatcccaa
2221 gtctgcaaac actcagtcca tctctcgaaa tcatgttgtg gaagtcagcg acagtggcct
2281 catcacaata gcaggtggga agtggaccac atatcgctcc atggcagaag atactgtgaa
2341 caaagctgtc aagcttcaca acttgaatgc aggacccagt aggaccgttg ggctcttcct
2401 tcaaggaggc aaagattgga gccccacact ctacatcagg cttgtccagg actatgggct
2461 tgagagtgag gttgcacaac acctggccaa aacctatggt gacaaggctt tcgacgtggc
2521 caaaatggca agtgtgactg gaaaacggtg gcctgttgtt ggagtgcgtc ttgtgtcaga
2581 atttccgtac atcgaagcag aggtgaaata tgggattaag gagtacgcat gcaccgctgt
2641 tgacatgatt tcacgtcgca cccgcctggc ctttcttaat gttcaggctg cagaggaagc
2701 cctgcccaag attgttgagt taatgggaag agaattgaac tggagtgaat tgaggaaaca
2761 ggaagaactt gaaacagcca cgaggtttct gtactatgaa atgggctata agtctcgaac
2821 agaacaactc acagatagta ctgaaatcag cctgctgcct ccagacatcg attatgatgt
2881 tcctgattat gcaagcttaa tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
2941 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
3001 cgagggcgat gccacctacg gcaagctgac cctgaagttc atcagcacca ccggcaagct
3061 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
3121 ctaccccgac cacatgaagc ggcacgactt cttcaagtcc gccatgcccg aaggctacgt
3181 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
3241 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
3301 cggcaacatc ctggggcaca agctggagta caactacaac tgccacaacg tctatatcat
3361 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
3421 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
3481 gctgctgccc gacaaccact acctgagcac ctgctccgcc ctgagcaaag accccaacga
3541 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
3601 ggacgagctg tacaagtaag aaggatccgg ctgcctcgcg cgtttcggtg atgacggtga
3661 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
3721 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
3781 gaggtcgact ctagtccccg cggtggcaga tctggaaggt gctgaggtac gatgagaccc
3841 gcaccaggtg cagaccctgc gagtgtggcg gtaaacatat taggaaccag cctgtgatgc
3901 tggatgtgac cgaggagctg aggcccgatc acttggtgct ggcctgcacc cgcgctgagt
3961 ttggctctag cgatgaagat acagattgag gtactgaaat gtgtgggcgt ggcttaaggg
4021 tgggaaagaa tatataaggt gggggtctta tgtagttttg tatctgtttt gcagcagccg
4081 ccgccgccat gagcaccaac tcgtttgatg gaagcattgt gagctcatat ttgacaacgc
4141 gcatgccccc atgggccggg gtgcgtcaga atgtgatggg ctccagcatt gatggtcgcc
4201 ccgtcctgcc cgcaaactct actaccttga cctacgagac cgtgtctgga acgccgttgg
4261 agactgcagc ctccgccgcc gcttcagccg ctgcagccac cgcccgcggg attgtgactg
4321 actttgcttt cctgagcccg cttgcaagca gtgcagcttc ccgttcatcc gcccgcgatg
4381 acaagttgac ggctcttttg gcacaattgg attctttgac ccgggaactt aatgtcgttt
4441 ctcagcagct gttggatctg cgccagcagg tttctgccct gaaggcttcc tcccctccca
4501 atgcggttta aaacataaat aaaaaaccag actctgtttg gatttggatc aagcaagtgt
4561 cttgctgtct ttatttaggg gttttgcgcg cgcggtaggc ccgggaccag cggtctcggt
4621 cgttgagggt cctgtgtatt ttttccagga cgtggtaaag gtgactctgg atgttcagat
4681 acatgggcat aagcccgtct ctggggtgga ggtagcacca ctgcagagct tcatgctgcg
4741 gggtggtgtt gtagatgatc cagtcgtagc aggagcgctg ggcgtggtgc ctaaaaatgt
4801 ctttcagtag caagctgatt gccaggggca ggcccttggt gtaagtgttt acaaagcggt
4861 taagctggga tgggtgcata cgtggggata tgagatgcat cttggactgt atttttaggt
4921 tggctatgtt cccagccata tccctccggg gattcatgtt gtgcagaacc accagcacag
4981 tgtatccggt gcacttggga aatttgtcat gtagcttaga aggaaatgcg tggaagaact
5041 tggagacgcc cttgtgacct ccaagatttt ccatgcattc gtccataatg atggcaatgg
5101 gcccacgggc ggcggcctgg gcgaagatat ttctgggatc actaacgtca tagttgtgtt
5161 ccaggatgag atcgtcatag gccattttta caaagcgcgg gcggagggtg ccagactgcg
5221 gtataatggt tccatccggc ccaggggcgt agttaccctc acagatttgc atttcccacg
5281 ctttgagttc agatgggggg atcatgtcta cctgcggggc gatgaagaaa acggtttccg
5341 gggtagggga gatcagctgg gaagaaagca ggttcctgag cagctgcgac ttaccgcagc
5401 cggtgggccc gtaaatcaca cctattaccg gctgcaactg gtagttaaga gagctgcagc
5461 tgccgtcatc cctgagcagg ggggccactt cgttaagcat gtccctgact cgcatgtttt
5521 ccctgaccaa atccgccaga aggcgctcgc cgcccagcga tagcagttct tgcaaggaag
5581 caaagttttt caacggtttg agaccgtccg ccgtaggcat gcttttgagc gtttgaccaa
5641 gcagttccag gcggtcccac agctcggtca cctgctctac ggcatctcga tccagcatat
5701 ctcctcgttt cgcgggttgg ggcggctttc gctgtacggc agtagtcggt gctcgtccag
5761 acgggccagg gtcatgtctt tccacgggcg cagggtcctc gtcagcgtag tctgggtcac
5821 ggtgaagggg tgcgctccgg gctgcgcgct ggccagggtg cgcttgaggc tggtcctgct
5881 ggtgctgaag cgctgccggt cttcgccctg cgcgtcggcc aggtagcatt tgaccatggt
5941 gtcatagtcc agcccctccg cggcgtggcc cttggcgcgc agcttgccct tggaggaggc
6001 gccgcacgag gggcagtgca gacttttgag ggcgtagagc ttgggcgcga gaaataccga
6061 ttccggggag taggcatccg cgccgcaggc cccgcagacg gtctcgcatt ccacgagcca
6121 ggtgagctct ggccgttcgg ggtcaaaaac caggtttccc ccatgctttt tgatgcgttt
6181 cttacctctg gtttccatga gccggtgtcc acgctcggtg acgaaaaggc tgtccgtgtc
6241 cccgtataca gacttgagag gcctgtcctc gaccgatgcc cttgagagcc ttcaacccag
6301 tcagctcctt ccggtgggcg cggggcatga ctatcgtcgc cgcacttatg actgtcttct
6361 ttatcatgca actcgtagga caggtgccgg cagcgctctg ggtcattttc ggcgaggacc
6421 gctttcgctg gagcgcgacg atgatcggcc tgtcgcttgc ggtattcgga atcttgcacg
6481 ccctcgctca agccttcgtc actggtcccg ccaccaaacg tttcggcgag aagcaggcca
6541 ttatcgccgg catggcggcc gacgcgctgg gctacgtctt gctggcgttc gcgacgcgag
6601 gctggatggc cttccccatt atgattcttc tcgcttccgg cggcatcggg atgcccgcgt
6661 tgcaggccat gctgtccagg caggtagatg acgaccatca gggacagctt caaggccagc
6721 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
6781 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
6841 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
6901 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
6961 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
7021 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
7081 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
7141 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
7201 ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
7261 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
7321 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
7381 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
7441 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
7501 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
7561 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
7621 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
7681 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
7741 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
7801 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
7861 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
7921 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
7981 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
8041 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
8101 gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc gcgccacata
8161 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
8221 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
8281 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
8341 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
8401 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
8461 aaaataaaca aataggggtt ccgcgcactt ttcggggaaa tgtgccacct gacgtctaag
8521 aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc
8581 ttcaattaat taagctag
//
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