| 质粒名称: | PCS2-FlipGFP(Casp3 cleavage seq) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 124432 |
| 存储实验室: | Xiaokun Shu |
|
载体骨架基本信息 |
|
| 载体名称: | pCS2 |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 斑马鱼（表达） |
| 筛选标记: | -- |
| 载体抗性: | 氨苄霉素100 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | FlipGFP(Casp3 cleavage seq) |
| 物种来源: | Synthetic |
| 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: | PCS2-FlipGFP(Casp3 cleavage seq) was a gift from Xiaokun Shu (Addgene plasmid # 124432 ; http://n2t.net/addgene:124432 ; RRID:Addgene_124432) |
| 参考文献部分: | Designing a green fluorogenic protease reporter by flipping a beta strand of GFP for imaging apoptosis in animals. Zhang Q, Schepis A, Huang H, Yang J, Ma W, Torra J, Zhang SQ, Yang L, Wu H, Nonell S, Dong Z, Kornberg TB, Coughlin SR, Shu X. J Am Chem Soc. 2019 Mar 1. doi: 10.1021/jacs.8b13042. 10.1021/jacs.8b13042 PubMed 30821975 |
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
质粒图谱.png)
载体简介
载体序列
LOCUS PCS2-FlipGFP(Casp3 cleavage seq) 5145 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses FlipGFP (caspase-3 cleavage sequence) protease reporter in
zebrafish (verified).
ACCESSION .
VERSION .
KEYWORDS PCS2-FlipGFP(Casp3 cleavage seq)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5145)
AUTHORS Zhang Q, Schepis A, Huang H, Yang J, Ma W, Torra J, Zhang SQ, Yang
L, Wu H, Nonell S, Dong Z, Kornberg TB, Coughlin SR, Shu X
TITLE Designing a green fluorogenic protease reporter by flipping a beta
strand of GFP for imaging apoptosis in animals.
JOURNAL J Am Chem Soc. 2019 Mar 1. doi: 10.1021/jacs.8b13042.
PUBMED 30821975
REFERENCE 2 (bases 1 to 5145)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5145
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 166..1143
/label=CMV IE94 promoter
/note="enhancer/promoter region of simian cytomegalovirus
major immediate early transcription unit IE94"
promoter 1178..1196
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 1178..1195
/label=SP6
/note="SP6 promoter, forward primer"
regulatory 1228..1237
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1630..1641
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
CDS 1633..1686
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
promoter complement(2318..2335)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
polyA_signal 2340..2474
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2364..2383)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2418..2437
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind complement(2587..2607)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(2587..2605)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(2626..2642)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2626..2642)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(2639..2661)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 2650..2666
/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(2674..2704)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2719..2740
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2857..2874)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3028..3616)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3108..3127)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(3787..4647)
/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 4410..4429
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(4648..4752)
/gene="bla"
/label=AmpR promoter
rep_origin complement(4778..88)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4915..4936)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5127..1
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
61 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
121 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg
181 cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg
241 cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg
301 cggacttggc accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg
361 gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat
421 attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata
481 taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt
541 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc
601 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc
661 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc
721 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc
781 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg
841 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg
901 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc
961 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc
1021 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg
1081 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg
1141 aaccgccatt ctgcctgggg acgtcggagc aagcttgatt taggtgacac tatagaatac
1201 aagctacttg ttctttttgc aggatccgcc accatggacc tgcctgacga ccactacctg
1261 tccacccaga ccatcctgtc caaggacctg aactccggac tcagatctgg cagcggtctc
1321 gagatggaag ttagcgctct ggaaaaagaa gtgtctgcac tcgagaaaga agtaagtgcc
1381 cttgagaagg aggtgtccgc actcgagaag gaggtcagcg ccctggaaaa ggaaaagcga
1441 gaccatatgg ttttgcttga gtatgttaca gcggctggca ttaccgatgc atcaggtgat
1501 gaagttgatg gtggcggtgg caaggtgtcc gccctgaagg aaaaagtaag cgcactgaaa
1561 gaaaaggtga gcgcgctgaa ggagaaagtg agcgccctga aagagaaagt ctctgccctt
1621 aaggaggata tcgagggcag aggaagtctg ctaacatgcg gtgacgtcga ggagaatcct
1681 ggcccaaagc ttgccaccat gcgcaaaggc gaagaactgt ttaccggcat tgtgccgatt
1741 ctggtggaac tggatggcga tgtgaacggc cataaatttt ttgtgcgcgg cgaaggcgaa
1801 ggcgatgcga ccattggcaa actgagcctg aaatttattt gcaccaccgg caaactgccg
1861 gtgccgtggc cgaccctggt gaccaccctg acctatggcg tgcagtgctt tagccgctat
1921 ccggatcata tgaaacgcca tgattttttt aaaagcgcga tgccggaagg ctatgtgcag
1981 gaacgcacca tttattttaa agatgatggc acctataaaa cccgcgcgga agtgaaattt
2041 gaaggcgata ccctggtgaa ccgcattgaa ctgaaaggca ttgattttaa agaagatggc
2101 aacattctgg gccataaact ggaatataac tttaacagcc ataaagtgta tattaccgcg
2161 gataaacaga acaacggcat taaagcgaac tttaccattc gccataacgt ggaagatggc
2221 agcgtgcagc tggcggatca ttatcagcag aacaccccga ttggcgatgg cccggttctt
2281 cttccttaag aattcaaggc ctctcgagcc tctagaacta tagtgagtcg tattacgtag
2341 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
2401 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
2461 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
2521 tgtgggaggt tttttaattc gcggccgcgg cgccaatgca ttgggcccgg tacccagctt
2581 ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc
2641 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggga gcataaagtg
2701 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
2761 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
2821 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
2881 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
2941 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
3001 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
3061 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
3121 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
3181 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
3241 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
3301 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
3361 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
3421 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
3481 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
3541 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
3601 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
3661 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
3721 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
3781 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
3841 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
3901 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
3961 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
4021 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
4081 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
4141 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
4201 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
4261 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
4321 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
4381 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
4441 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
4501 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
4561 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
4621 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
4681 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
4741 aggggttccg cgcacatttc cccgaaaagt gccacctaaa ttgtaagcgt taatattttg
4801 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
4861 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
4921 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
4981 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
5041 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
5101 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aagga
//
