| 质粒名称: | GW1-Mito-pHRed |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 31474 |
| 存储实验室: | Gary Yellen |
|
载体骨架基本信息 |
|
| 载体名称: | GW1 |
| 载体出品公司: | British Biotechnology |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5300 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: | 未知 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Cox8-tagged pHRed |
| 物种来源: | 合成 |
| 插入大小: | 1100 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 4xCox8 (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | XbaI(未破坏) |
| 3' 克隆位点: | NheI(未破坏) |
| 5' 测序引物 | CACAGCAATCGATCCATG |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | GW1-Mito-pHRed was a gift from Gary Yellen (Addgene plasmid # 31474 ; http://n2t.net/addgene:31474 ; RRID:Addgene_31474) |
| 参考文献部分: | Imaging Intracellular pH in Live Cells with a Genetically Encoded Red Fluorescent Protein Sensor. Tantama M, Hung YP, Yellen G. J Am Chem Soc. 2011 Jul 6. 133(26):10034-7. 10.1021/ja202902d PubMed 21631110 |
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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 GW1-Mito-pHRed 6927 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS GW1-Mito-pHRed
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6927)
AUTHORS Tantama M, Hung YP, Yellen G
TITLE Imaging Intracellular pH in Live Cells with a Genetically Encoded
Red Fluorescent Protein Sensor.
JOURNAL J Am Chem Soc. 2011 Jul 6. 133(26):10034-7.
PUBMED 21631110
REFERENCE 2 (bases 1 to 6927)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6927
/organism="synthetic DNA construct"
/mol_type="other DNA"
polyA_signal 1420..1554
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1457..1476)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1511..1530
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(1577..1593)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1577..1593)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1590..1612)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1601..1617
/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(1625..1655)
/label=lac promoter
/note="promoter for the E. coli lac operon"
primer_bind complement(1807..1824)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1978..2566)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(2058..2077)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2737..3597)
/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 3360..3379
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin complement(3654..4167)
/direction=LEFT
/label=M13 ori
/note="M13 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3849..3870)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 4061..4080
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4339..4357
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(4395..4417)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 4517..4536
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4604..4623)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 4768..4790
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 4782..4799
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 4783..4799
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
enhancer 5365..5744
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 5745..5948
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 5899..5919
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 5945..5969
/label=LNCX
/note="Human CMV promoter, forward primer"
ORIGIN
1 aagcttcccg gccacagcaa tcgatccatg acacaaccag tgacaagcag tcccccagtg
61 aaaccgcatc cgcatccgag tccgataccg ataaagattc tgaatcggag tgagtgccgc
121 gtccgagagc gttccctgtc cacgtccacc atcggcggag caggtgtgcc tgaggcctct
181 agaccaccat gagcgtgctg acaccattgt tgctcagagg cctcactggt tcagccaggc
241 ggctgccagt cccaagagct aagatccata gtctgggaga tccaatgtca gttctcactc
301 cccttttgct taggggactg accggaagcg ctcggcggtt gcctgtccca cgcgccaaga
361 ttcactccct gggcgaccct atgagtgtcc ttactcctct tttgctgaga ggcttgacag
421 ggagcgctcg gaggctccca gtgcctcgcg caaagatcca ttctctcgga gatcccatgt
481 ccgtcctcac tcctctgctg ctgcgcgggc ttacaggttc cgctcgccgc ctgcctgttc
541 ctcgggcaaa gatacacagt ctgggcgatc ctcaccggcg ctatctcatg ggcggtgcaa
601 gcggcggcgc aagcggtggt atggtttccg ttatcgcaaa gcaaatgacc tataaagttt
661 atatgtctgg caccgttaac ggccactact tcgaggttga aggtgacggc aagggcaagc
721 cttacgaggg cgagcagacc gttaagctga ccgttaccaa aggcggccca ctgcctttcg
781 cctgggatat cctgtcccct cagctccaat acggctccat ccctttcacc aaataccccg
841 aggacatccc tgactacttt aagcagagct tcccagaggg ctacacctgg gagcgctcta
901 tgaacttcga ggacggcgca gtttgtaccg tttccaacga cagctccatc cagggtaact
961 gcttcatcta caacgttaag atttccggcg agaacttccc acctaacggc ccagttatgc
1021 agaagaaaac ccaaggctgg gagcccagca ccgaacgcct gtttgcccgc gacggtatgc
1081 tgattggtaa cgattatatg gccctgaaac tggaaggcgg cggtcactac ctgtgcgagt
1141 tcaagagcac ttacaaggct aaaaagcctg ttcgcatgcc tggccgccac gaaatcgacc
1201 gcaagctgga tgttacctct cacaaccgcg actacaccag cgttgagcag tgtgaaatct
1261 ctatcgcccg ccactctctg ctgggttgag ctagctgagt cgacgaattc attgatcata
1321 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
1381 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
1441 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
1501 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccgtacc
1561 gagctcgcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
1621 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
1681 aactacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
1741 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
1801 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
1861 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
1921 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
1981 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
2041 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
2101 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
2161 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
2221 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
2281 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
2341 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
2401 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
2461 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
2521 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
2581 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
2641 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
2701 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
2761 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
2821 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
2881 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
2941 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
3001 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
3061 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
3121 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
3181 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
3241 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
3301 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
3361 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
3421 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
3481 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
3541 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
3601 cttccttttt caataagcgg ccgcggccat gccggccact agtctcgagg gccaccccgg
3661 ttgataatca gaaaagcccc aaaaacagga agattgtata agcaaatatt taaattgtaa
3721 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
3781 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
3841 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
3901 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
3961 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
4021 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
4081 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
4141 cgcttaatgc gccgctacag ggcgcgtggg gatccctcga gttattattg aagcatttat
4201 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
4261 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
4321 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
4381 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
4441 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
4501 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgaagacg
4561 tcgcctcctc actacttctg gaatagctca gaggccgagg cggcctcggc ctctgcataa
4621 ataaaaaaaa ttagtcaggg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
4681 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
4741 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
4801 cccgtgtcaa ggacggtgac tgcagtgaat aataaaatgt gtgtttgtcc gaaatacgcg
4861 ttttgagatt tctgtcgccg actaaattca tgtcgcgcga tagtggtgtt tatcgccgat
4921 agagatggcg atattggaaa aatcgatatt tgaaaatatg gcatattgaa aatgtcgccg
4981 atgtgagttt ctgtgtaact gatatcgcca tttttccaaa agtgattttt gggcatacgc
5041 gatatctggc gatagcgctt atatcgttta cgggggatgg cgatagacga ctttggtgac
5101 ttgggcgatt ctgtgtgtcg caaatatcgc agtttcgata taggtgacag acgatatgag
5161 gctatatcgc cgatagaggc gacatcaagc tggcacatgg ccaatgcata tcgatctata
5221 cattgaatca atattggcca ttagccatat tattcattgg ttatatagca taaatcaata
5281 ttggctattg gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct
5341 catgtccaac attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
5401 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
5461 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
5521 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
5581 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
5641 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
5701 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
5761 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
5821 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
5881 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
5941 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
6001 tccatagaag acaccgggac cgatccagcc tccgcggccg ggaacggtgc attggaacgc
6061 ggattccccg tgccaagagt gacgtaagta ccgcctatag agtctatagg cccaccccct
6121 tggcttctta tgcatgctat actgtttttg gcttggggtc tatacacccc cgcttcctca
6181 tgttataggt gatggtatag cttagcctat aggtgtgggt tattgaccat tattgaccac
6241 tcccctattg gtgacgatac tttccattac taatccataa catggctctt tgccacaact
6301 ctctttattg gctatatgcc aatacactgt ccttcagaga ctgacacgga ctctgtattt
6361 ttacaggatg gggtctcatt tattatttac aaattcacat atacaacacc accgtcccca
6421 gtgcccgcag tttttattaa acataacgtg ggatctccac gcgaatctcg ggtacgtgtt
6481 ccggacatgg gctcttctcc ggtagcggcg gagcttctac atccgagccc tgctcccatg
6541 cctccagcga ctcatggtcg ctcggcagct ccttgctcct aacagtggag gccagactta
6601 ggcacagcac gatgcccacc accaccagtg tgccgcacaa ggccgtggcg gtagggtatg
6661 tgtctgaaaa tgagctcggg gagcgggctt gcaccgctga cgcatttgga agacttaagg
6721 cagcggcaga agaagatgca ggcagctgag ttgttgtgtt ctgataagag tcagaggtaa
6781 ctcccgttgc ggtgctgtta acggtggagg gcagtgtagt ctgagcagta ctcgttgctg
6841 ccgcgcgcgc caccagacat aatagctgac agactaacag actgttcctt tccatgggtc
6901 ttttctgcag tcaccgtcct tgacacg
//
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