| 质粒名称: | pQE-30-HyPer3 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 42132 |
| 存储实验室: | Vsevolod Belousov |
|
载体骨架基本信息 |
|
| 载体名称: | pQE-30 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 3425 bp |
| 完整质粒大小: | 4859 bp |
| 载体类型: | 细菌表达 |
| 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: | HYPER 3 |
| 物种来源: | Synthetic |
| 插入大小: | 1434 bp |
| 启动子 | T5 |
| 标签/融合蛋白: | 6His（主干上的N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI(未破坏) |
| 3' 克隆位点: | HindIII (未破坏) |
| 5' 测序引物 | aggagaaattaactatgagagg |
| 3' 测序引物 | ccagatggagttctgaggtc |
|
文献引用方法 |
|
| 材料和方法部分: | pQE-30-HyPer3 was a gift from Vsevolod Belousov (Addgene plasmid # 42132 ; http://n2t.net/addgene:42132 ; RRID:Addgene_42132) |
| 参考文献部分: | HyPer-3: a genetically encoded H2O2 probe with improved performance for ratiometric and fluorescence lifetime imaging. Bilan DS, Pase L, Joosen L, Gorokhovatsky AY, Ermakova YG, Gadella TW, Grabher C, Schultz C, Lukyanov S, Belousov VV. ACS Chem Biol. 2012 Dec 20. 10.1021/cb300625g PubMed 23256573 |
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 pQE-30-HyPer3 4859 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Genetically encoded H2O2 probe for ratiometric and fluorescence
lifetime imaging.
ACCESSION .
VERSION .
KEYWORDS pQE-30-HyPer3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4859)
AUTHORS Bilan DS, Pase L, Joosen L, Gorokhovatsky AY, Ermakova YG, Gadella
TW, Grabher C, Schultz C, Lukyanov S, Belousov VV
TITLE HyPer-3: a genetically encoded H2O2 probe with improved performance
for ratiometric and fluorescence lifetime imaging.
JOURNAL ACS Chem Biol. 2012 Dec 20.
PUBMED 23256573
REFERENCE 2 (bases 1 to 4859)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4859
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 10..54
/label=T5 promoter
/note="bacteriophage T5 promoter for E. coli RNA
polymerase, with embedded lac operator"
protein_bind 30..46
/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)."
protein_bind 62..78
/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)."
RBS 97..108
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 127..144
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 151..1584
/codon_start=1
/product="improved fluorescent indicator for intracellular
H2O2 (Bilan et al., 2013)"
/label=HyPer-3
/translation="EMASQQGETMSGPLHIGLIPTVGPYLLPHIIPMLYQTFPKLEMYL
HEAQTHQLLAQLDSGKLDCVILALVKESEAFIEVPLFDEPMLLAIYEDHPWANRECVPM
ADLAGEKLLMLEDGHCLRDQAMSAGYNSDNVYIMADKQKNGIKANFKIRHNVEDGSVQL
ADHYQQNTPIGDGPVLLPDNHYLSFQSVLSKDPNEKRDHMVLLEFVTAAGITLGMDELY
NVDGGSGGTGSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKLICTT
GKLPVPWPTLVTTLGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTR
AEVKFEGDTLVNRIELKGIGFKEDGNILGHKLEYNGTGFCFEAGADEDTHFRATSLETL
RNMVAAGSGITLLPALAVPPERKRDGVVYLPCIKPEPRRTIGLVYRPGSPLRSRYEQLA
EAIRARMDGHFDKVLKQAV"
primer_bind 756..777
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(886..907)
/label=EGFP-N
/note="EGFP, reverse primer"
primer_bind complement(1147..1166)
/label=EXFP-R
/note="For distinguishing EGFP variants, reverse primer"
terminator 1606..1700
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS 1744..2403
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
primer_bind complement(1810..1829)
/label=CAT-R
/note="Chloramphenicol resistance gene, reverse primer"
terminator 2468..2554
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind complement(2601..2623)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 2709..2849
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 2764..2783
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(2864..2881)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3035..3623)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3115..3134)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3794..4654)
/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 4417..4436
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4655..4759)
/gene="bla"
/label=AmpR promoter
primer_bind 4827..4845
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
121 ggatcgcatc accatcacca tcacggatcc gagatggcaa gccagcaggg cgagacgatg
181 tccggaccgc tgcacattgg tttgattccc acagttggac cgtacctgct accgcatatt
241 atccctatgc tgtaccagac ctttccaaag ctggaaatgt atctgcatga agcacagacc
301 caccagttac tggcgcaact ggacagcggc aaactcgatt gcgtgatcct cgcactggtg
361 aaagagagcg aagcattcat tgaagtgccg ttgtttgatg agccaatgtt gctggctatc
421 tatgaagatc acccgtgggc gaaccgcgaa tgcgtaccga tggccgatct ggcaggggaa
481 aaactgctga tgctggaaga tggtcactgt ttgcgcgatc aggcaatgtc cgccggctac
541 aacagcgaca acgtctatat catggccgac aagcagaaga acggcatcaa ggccaacttc
601 aagatccgcc acaacgtcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
661 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cttccagtcc
721 gtcctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
781 gccgccggga tcactctcgg catggacgag ctgtacaacg tggatggcgg tagcggtggc
841 accggcagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
901 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
961 ggcaagctga ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc
1021 ctcgtgacca ccctcggcta cggcctgaag tgcttcgccc gctaccccga ccacatgaag
1081 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
1141 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
1201 gtgaaccgca tcgagctgaa gggcatcggc ttcaaggagg acggcaacat cctggggcac
1261 aagctggagt acaacggcac cggtttctgt tttgaagccg gggcggatga agatacacac
1321 ttccgcgcga ccagcctgga aactctgcgc aacatggtgg cggcaggtag cgggatcact
1381 ttactgccag cgctggctgt gccgccggag cgcaaacgcg atggggttgt ttatctgccg
1441 tgcattaagc cggaaccacg ccgcactatt ggtctggttt atcgtcctgg ctcaccgctg
1501 cgcagccgct atgagcagct ggcagaggcc atccgcgcaa gaatggatgg ccatttcgat
1561 aaagttttaa aacaggcggt ttaaaagctt aattagctga gcttggactc ctgttgatag
1621 atccagtaat gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg
1681 cgttttttat tggtgagaat ccaagctagc ttggcgagat tttcaggagc taaggaagct
1741 aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa
1801 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
1861 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
1921 attcacattc ttgcccgcct gatgaatgct catccggaat ttcgtatggc aatgaaagac
1981 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
2041 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
2101 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
2161 gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
2221 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
2281 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
2341 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg
2401 taattttttt aaggcagtta ttggtgccct taaacgcctg gggtaatgac tctctagctt
2461 gaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
2521 tttgtcggtg aacgctctcc tgagtaggac aaatccgccc tctagagctg cctcgcgcgt
2581 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
2641 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
2701 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact
2761 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
2821 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
2881 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
2941 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
3001 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
3061 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
3121 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
3181 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
3241 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
3301 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
3361 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
3421 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
3481 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
3541 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
3601 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
3661 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
3721 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
3781 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
3841 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
3901 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
3961 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
4021 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
4081 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
4141 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
4201 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
4261 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
4321 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
4381 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
4441 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
4501 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
4561 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
4621 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
4681 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
4741 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
4801 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcac
//
