| 质粒名称: | pcDNA3.1 iCasper T2A HO1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 64278 |
| 存储实验室: | Xiaokun Shu |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5360 bp |
| 完整质粒大小: | 7920 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Infrared fluorescent executioner caspase reporter (iCasper) T2A heme oxygenase-1:,iCasper T2A HO1 |
| 物种来源: | 合成 |
| 插入大小: | 2560 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 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: | HindIII(未破坏) |
| 3' 克隆位点: | XbaI (未破坏) |
| 5' 测序引物 | TAATACGACTCACTATAGGG |
| 3' 测序引物 | TAGAAGGCACAGTCGAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pcDNA3.1 iCasper T2A HO1 was a gift from Xiaokun Shu (Addgene plasmid # 64278 ; http://n2t.net/addgene:64278 ; RRID:Addgene_64278) |
| 参考文献部分: | Rationally designed fluorogenic protease reporter visualizes spatiotemporal dynamics of apoptosis in vivo. To TL, Piggott BJ, Makhijani K, Yu D, Jan YN, Shu X. Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3338-43. doi: 10.1073/pnas.1502857112. Epub 2015 Mar 2. 10.1073/pnas.1502857112 PubMed 25733847 |
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 pcDNA3.1 iCasper T2A HO1 7922 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses infrared fluorescent executioner caspase reporter
(iCasper) plus human heme oxygenase 1.
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1 iCasper T2A HO1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7922)
AUTHORS To TL, Piggott BJ, Makhijani K, Yu D, Jan YN, Shu X
TITLE Rationally designed fluorogenic protease reporter visualizes
spatiotemporal dynamics of apoptosis in vivo.
JOURNAL Proc Natl Acad Sci U S A. 2015 Mar 17;112(11):3338-43. doi:
10.1073/pnas.1502857112. Epub 2015 Mar 2.
PUBMED 25733847
REFERENCE 2 (bases 1 to 7922)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7922
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 769..789
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 863..882
/label=T7
/note="T7 promoter, forward primer"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 923..1564
/codon_start=1
/product="fragment of superfolder GFP consisting of the
first 10 beta-strands"
/label=GFP(1-10)
/note="associates with GFP11, the 11th beta-strand, to
reconstitute fluorescent GFP (Cabantous et al., 2005;
Kamiyama et al., 2016)"
/translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
KYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEK"
CDS 2507..2554
/codon_start=1
/product="11th beta-strand of superfolder GFP"
/label=GFP11
/note="associates with GFP(1-10), consisting of the first
10 beta-strands, to reconstitute fluorescent GFP (Cabantous
et al., 2005; Kamiyama et al., 2016)"
/translation="RDHMVLHEYVNAAGIT"
CDS 2564..2617
/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"
primer_bind complement(3516..3533)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 3522..3746
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 3792..4220
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3879..3898)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4089..4110
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(4229..4249)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 4234..4563
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 4414..4549
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4476..4495
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 4630..5424
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4684..4703)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 5294..5313
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 5598..5719
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5635..5654)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5689..5708
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(5768..5784)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5768..5784)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5781..5803)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5792..5808
/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(5816..5846)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5861..5882
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5999..6016)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6170..6755)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6250..6269)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6926..7786)
/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 7549..7568
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(7787..7891)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
901 gtttaaactt aagcttgcca ccatgtccaa aggagaagaa ctgtttaccg gtgttgtgcc
961 aattttggtt gaactcgatg gtgatgtcaa cggacataag ttctcagtga gaggcgaagg
1021 agaaggtgac gccaccattg gaaaattgac tcttaaattc atctgtacta ctggtaaact
1081 tcctgtacca tggccgactc tcgtaacaac gcttacgtac ggagttcagt gcttttcgag
1141 atacccagac catatgaaaa gacatgactt ttttaagtcg gctatgcctg aaggttacgt
1201 gcaagaaaga acaatttcgt tcaaagatga tggaaaatat aaaactagag cagttgttaa
1261 atttgaagga gatactttgg ttaaccgcat tgaactgaaa ggaacagatt ttaaagaaga
1321 tggtaatatt cttggacaca aactcgaata caattttaat agtcataacg tatacatcac
1381 tgctgataag caaaagaacg gaattaaagc gaatttcaca gtacgccata atgtagaaga
1441 tggcagtgtt caacttgccg accattacca acaaaacacc cctattggag acggtccggt
1501 acttcttcct gataatcact acctctcaac acaaacagtc ctgagcaaag atccaaatga
1561 aaaaggaggc agcggatcca ccactaacat tgcgccggct ctggacggtg cgtttcatcg
1621 tatcacttct tcatcctccc tgatgggcct gtgtgacgaa accgcgacta ttatccgtga
1681 gattactggc tacgaccgtg tgatggtagt acgtttcgat gaagagggta atgccgaaat
1741 tctgtccgaa cgtcgtcgtg cggacctgga agcgttcctg ggtaaccgct acccggcgtc
1801 tactattccg cagatcgctc gtcgcctgta cgaacataac cgtgttcgcc tgctggtaga
1861 tgtgaactat actccggttc cgctacagcc gcgcatcagc ccgctgaacg gtcgtgatct
1921 ggatatgtcc ctgtcttgcc tgcgctctat gtccccggtc caccagaaat acatgcagga
1981 catgggcgtt ggcgcgaccc tggtttgctc tctgatggtg tctggtcgtt tgtggggtct
2041 gatcgcttgc caccactacg aaccgcgctt cgttccgttc cacattcgcg ctgctggcga
2101 agcgctggcg gaaacttgtg cgaaccgcat cgcgacgctg gagagcggtg atgaagttga
2161 tggtgaattc tgtgaacgcg agcagatcca cctggcgggc tccattcagc cgcacggtat
2221 cctgctggct gtgaaagagc cagacaacgt ggtgatccag gcttctatca acgctgcgga
2281 gttcctgaac accaactctg ttgttggccg tccgctgcgt gacctgggcg gcgatctgcc
2341 tttgcagatc ctgccgcacc tgaacggccc gctgcacctg gctccggtga ccctgcgttg
2401 taccgtgggt tctccgccgc gccgtgtgga ctgtaccatt caccgtccgt ctaacggcgg
2461 cctgatcgta gaactggaac cagcaaccaa gctcgaggga ggcagccggg accacatggt
2521 gctgcacgag tacgtgaacg ccgccggcat cacagggcgc gccgagggca gaggaagtct
2581 gctaacatgc ggtgacgtcg aggagaatcc tggcccaatg gaaagaccac agccagactc
2641 tatgccacag gatctgagcg aagccctcaa ggaggccacc aaggaagtgc acacccaggc
2701 cgagaacgct gagttcatga gaaacttcca gaaaggacag gttactcgcg acggattcaa
2761 gctggtgatg gctagcctct atcacatcta cgtggccctg gaggaggaaa ttgagagaaa
2821 caaggaatcc cccgtgttcg ctccagttta tttcccagag gagctgcatc ggaaggccgc
2881 cctggaacag gatctggctt tttggtatgg cccccgctgg caggaggtca ttccttatac
2941 ccccgctatg caacgctacg ttaagagact ccacgaggtg ggccgcacag aacctgagct
3001 gctcgtggcc catgcctaca cccgctacct gggcgacctg tcagggggtc aagttctgaa
3061 aaagatagcc caaaaggccc tggacctgcc ttcctctggc gaggggctgg ccttctttac
3121 tttccctaac attgcctcag ccacaaagtt caagcagctc taccggtcca ggatgaacag
3181 cctggaaatg accccagccg tgcggcagcg ggtcatagaa gaagccaaga ctgcctttct
3241 gctgaatatc cagctgtttg aggaactcca ggaactgctg acccatgaca ctaaagacca
3301 gtccccctcc agggccccag gactcagaca gagggcctca aacaaggttc aggactcagc
3361 ccccgtggaa acacccaggg gaaaaccccc tctcaataca agatctcaag cacccctgct
3421 gagatgggtc ctcaccctga gcttcctcgt cgctaccgtt gcagtggggc tctatgctat
3481 gtgatctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
3541 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
3601 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
3661 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
3721 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
3781 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
3841 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
3901 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
3961 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
4021 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
4081 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
4141 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
4201 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
4261 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
4321 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
4381 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
4441 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
4501 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
4561 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
4621 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
4681 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
4741 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
4801 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
4861 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
4921 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
4981 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
5041 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
5101 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
5161 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
5221 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
5281 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
5341 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
5401 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
5461 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
5521 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
5581 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
5641 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
5701 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
5761 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
5821 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
5881 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
5941 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
6001 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
6061 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
6121 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
6181 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
6241 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
6301 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
6361 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
6421 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
6481 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
6541 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
6601 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
6661 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
6721 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
6781 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
6841 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
6901 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
6961 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
7021 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
7081 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
7141 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
7201 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
7261 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
7321 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
7381 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
7441 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
7501 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
7561 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
7621 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
7681 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
7741 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
7801 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
7861 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
7921 tc
//
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