| 质粒名称: | Cyto-roGFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 49435 |
| 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: | 5970 bp |
| 载体修饰: | -- |
| 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: | roGFP2 |
| 物种来源: | -- |
| 插入大小: | 750 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 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: | Cyto-roGFP was a gift from Paul Schumacker (Addgene plasmid # 49435 ; http://n2t.net/addgene:49435 ; RRID:Addgene_49435) |
| 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 Cyto-roGFP 6883 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses thiol redox-sensitive ratiometric sensor roGFP in cytosol
of mammalian cells.
ACCESSION .
VERSION .
KEYWORDS Cyto-roGFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6883)
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 6883)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6883
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(108..127)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
primer_bind 455..476
/label=EGFP-C
/note="EGFP, forward primer"
polyA_signal 542..676
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(566..585)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 620..639
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(819..841)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin complement(3916..4504)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3996..4015)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4675..5535)
/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 5298..5317
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5536..5640)
/gene="bla"
/label=AmpR promoter
primer_bind 5708..5726
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
repeat_region 5754..5856
/label=ITR
/note="inverted terminal repeat of human adenovirus
serotype 5"
misc_signal 5944..6094
/label=Ad5 Psi
/note="packaging signal for adenovirus serotype 5"
enhancer 6123..6426
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 6427..6630
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 6581..6601
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
regulatory 6679..6688
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 6685..521
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
primer_bind complement(6730..6751)
/label=EGFP-N
/note="EGFP, reverse primer"
ORIGIN
1 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcggcacgac ttcttcaagt
61 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
121 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
181 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
241 actgccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
301 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
361 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acctgctccg
421 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
481 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc cacagcgggg
541 agatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
601 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
661 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
721 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
781 tccggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
841 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
901 tcagcgggtg ttggcgggtg tcggggcgca gccatgaggt cgactctagt ccccgcggtg
961 gcagatctgg aaggtgctga ggtacgatga gacccgcacc aggtgcagac cctgcgagtg
1021 tggcggtaaa catattagga accagcctgt gatgctggat gtgaccgagg agctgaggcc
1081 cgatcacttg gtgctggcct gcacccgcgc tgagtttggc tctagcgatg aagatacaga
1141 ttgaggtact gaaatgtgtg ggcgtggctt aagggtggga aagaatatat aaggtggggg
1201 tcttatgtag ttttgtatct gttttgcagc agccgccgcc gccatgagca ccaactcgtt
1261 tgatggaagc attgtgagct catatttgac aacgcgcatg cccccatggg ccggggtgcg
1321 tcagaatgtg atgggctcca gcattgatgg tcgccccgtc ctgcccgcaa actctactac
1381 cttgacctac gagaccgtgt ctggaacgcc gttggagact gcagcctccg ccgccgcttc
1441 agccgctgca gccaccgccc gcgggattgt gactgacttt gctttcctga gcccgcttgc
1501 aagcagtgca gcttcccgtt catccgcccg cgatgacaag ttgacggctc ttttggcaca
1561 attggattct ttgacccggg aacttaatgt cgtttctcag cagctgttgg atctgcgcca
1621 gcaggtttct gccctgaagg cttcctcccc tcccaatgcg gtttaaaaca taaataaaaa
1681 accagactct gtttggattt ggatcaagca agtgtcttgc tgtctttatt taggggtttt
1741 gcgcgcgcgg taggcccggg accagcggtc tcggtcgttg agggtcctgt gtattttttc
1801 caggacgtgg taaaggtgac tctggatgtt cagatacatg ggcataagcc cgtctctggg
1861 gtggaggtag caccactgca gagcttcatg ctgcggggtg gtgttgtaga tgatccagtc
1921 gtagcaggag cgctgggcgt ggtgcctaaa aatgtctttc agtagcaagc tgattgccag
1981 gggcaggccc ttggtgtaag tgtttacaaa gcggttaagc tgggatgggt gcatacgtgg
2041 ggatatgaga tgcatcttgg actgtatttt taggttggct atgttcccag ccatatccct
2101 ccggggattc atgttgtgca gaaccaccag cacagtgtat ccggtgcact tgggaaattt
2161 gtcatgtagc ttagaaggaa atgcgtggaa gaacttggag acgcccttgt gacctccaag
2221 attttccatg cattcgtcca taatgatggc aatgggccca cgggcggcgg cctgggcgaa
2281 gatatttctg ggatcactaa cgtcatagtt gtgttccagg atgagatcgt cataggccat
2341 ttttacaaag cgcgggcgga gggtgccaga ctgcggtata atggttccat ccggcccagg
2401 ggcgtagtta ccctcacaga tttgcatttc ccacgctttg agttcagatg gggggatcat
2461 gtctacctgc ggggcgatga agaaaacggt ttccggggta ggggagatca gctgggaaga
2521 aagcaggttc ctgagcagct gcgacttacc gcagccggtg ggcccgtaaa tcacacctat
2581 taccggctgc aactggtagt taagagagct gcagctgccg tcatccctga gcaggggggc
2641 cacttcgtta agcatgtccc tgactcgcat gttttccctg accaaatccg ccagaaggcg
2701 ctcgccgccc agcgatagca gttcttgcaa ggaagcaaag tttttcaacg gtttgagacc
2761 gtccgccgta ggcatgcttt tgagcgtttg accaagcagt tccaggcggt cccacagctc
2821 ggtcacctgc tctacggcat ctcgatccag catatctcct cgtttcgcgg gttggggcgg
2881 ctttcgctgt acggcagtag tcggtgctcg tccagacggg ccagggtcat gtctttccac
2941 gggcgcaggg tcctcgtcag cgtagtctgg gtcacggtga aggggtgcgc tccgggctgc
3001 gcgctggcca gggtgcgctt gaggctggtc ctgctggtgc tgaagcgctg ccggtcttcg
3061 ccctgcgcgt cggccaggta gcatttgacc atggtgtcat agtccagccc ctccgcggcg
3121 tggcccttgg cgcgcagctt gcccttggag gaggcgccgc acgaggggca gtgcagactt
3181 ttgagggcgt agagcttggg cgcgagaaat accgattccg gggagtaggc atccgcgccg
3241 caggccccgc agacggtctc gcattccacg agccaggtga gctctggccg ttcggggtca
3301 aaaaccaggt ttcccccatg ctttttgatg cgtttcttac ctctggtttc catgagccgg
3361 tgtccacgct cggtgacgaa aaggctgtcc gtgtccccgt atacagactt gagaggcctg
3421 tcctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
3481 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
3541 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
3601 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
3661 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc
3721 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat
3781 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
3841 agatgacgac catcagggac agcttcaagg ccagcaaaag gccaggaacc gtaaaaaggc
3901 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
3961 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
4021 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
4081 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
4141 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
4201 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
4261 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
4321 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
4381 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
4441 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
4501 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
4561 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
4621 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
4681 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
4741 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
4801 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
4861 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
4921 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
4981 tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
5041 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
5101 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
5161 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
5221 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
5281 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
5341 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
5401 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
5461 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
5521 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
5581 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
5641 cacttttcgg ggaaatgtgc cacctgacgt ctaagaaacc attattatca tgacattaac
5701 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa ttaattaagc tagcatcatc
5761 aataatatac cttattttgg attgaagcca atatgataat gagggggtgg agtttgtgac
5821 gtggcgcggg gcgtgggaac ggggcgggtg acgtagtagt gtggcggaag tgtgatgttg
5881 caagtgtggc ggaacacatg taagcgacgg atgtggcaaa agtgacgttt ttggtgtgcg
5941 ccggtgtaca caggaagtga caattttcgc gcggttttag gcggatgttg tagtaaattt
6001 gggcgtaacc gagtaagatt tggccatttt cgcgggaaaa ctgaataaga ggaagtgaaa
6061 tctgaataat tttgtgttac tcatagcgcg taatatttgt ctagggagat cagcctgcag
6121 gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
6181 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
6241 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
6301 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
6361 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
6421 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
6481 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
6541 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
6601 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat ggtaccgcgg
6661 gcccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
6721 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
6781 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatcag caccaccggc
6841 aagctgcccg tgccctggcc caccctcgtg accaccctga cct
//
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