| 质粒名称: | p53-TRE-mStrawberry reporter |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 158684 |
| 存储实验室: | Ravid Straussman |
|
载体骨架基本信息 |
|
| 载体名称: | pLKO.1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 7728 bp |
| 完整质粒大小: | 7959 bp |
| 载体类型: | 哺乳细胞表达, Lentiviral |
| 筛选标记: | Blasticidin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Strawberry |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 231 bp |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | p53-TRE-mStrawberry reporter was a gift from Ravid Straussman (Addgene plasmid # 158684 ; http://n2t.net/addgene:158684 ; RRID:Addgene_158684) |
| 参考文献部分: | Visual barcodes for clonal-multiplexing of live microscopy-based assays. Kaufman T, Nitzan E, Firestein N, Ginzberg MB, Iyengar S, Patel N, Ben-Hamo R, Porat Z, Hunter J, Hilfinger A, Rotter V, Kafri R, Straussman R. Nat Commun. 2022 May 18;13(1):2725. doi: 10.1038/s41467-022-30008-0. 10.1038/s41467-022-30008-0 PubMed 35585055 |
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 p53-TRE-mStrawberry reporter 7959 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION This plasmid is a p53 pathway reporter. It has two copies of
wild-type p53 binding sites promoter driving the expression of
mStrawberry-pGK-BSD.
ACCESSION .
VERSION .
KEYWORDS p53-TRE-mStrawberry reporter
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7959)
TITLE Straussman Lab plasmids
REFERENCE 2 (bases 1 to 7959)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7959
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1..711
/codon_start=1
/product="monomeric derivative of DsRed fluorescent
protein"
/label=mStrawberry
/note="mammalian codon-optimized"
/translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILTPNFTYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKMRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYIVGIKLDITSHNEDYTIVELYERA
EGRHSTGGMDELYK"
primer_bind complement(146..164)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 414..433
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 753..870
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 929..1429
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 1348..1366
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 1441..1836
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
misc_feature 1872..2460
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(1925..1945)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(2343..2354)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 2532..2765
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 2843..2964
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2880..2899)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 2934..2953
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 3004..3139
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3066..3085
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(3159..3178)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3160..3178)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3188..3205)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(3188..3204)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3197..3219)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 3346..3801
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(3433..3452)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3643..3664
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 3827..3931
/gene="bla"
/label=AmpR promoter
CDS 3932..4792
/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 complement(4150..4169)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4963..5551
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 5452..5471
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5705..5722
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5839..5860
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5875..5905
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5913..5929
/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)."
primer_bind 5918..5940
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5937..5953
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5937..5953
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 5972..5992
/label=T3
/note="T3 promoter, forward primer"
promoter 5974..5992
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 6020..6246
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 6247..6427
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 6474..6599
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 7092..7325
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 7510..7554
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
ORIGIN
1 atggtgagca agggcgagga gaataacatg gccatcatca aggagttcat gcgcttcaag
61 gtgcgcatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
121 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
181 ttcgcctggg acatcctaac ccccaacttc acctacggct ccaaggccta cgtgaagcac
241 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
301 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
361 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
421 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
481 gccctgaagg gcgagatcaa gatgaggctg aagctgaagg acggcggcca ctacgacgct
541 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacatcgtc
601 ggcatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga actgtacgaa
661 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta agaattctag
721 atcttgagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
781 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
841 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
901 ctttggcgcc ggctcgaggg ggcccgggtt gcgccttttc caaggcagcc ctgggtttgc
961 gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg accctgggtc
1021 tcgcacattc ttcacgtccg ttcgcagcgt cacccggatc ttcgccgcta cccttgtggg
1081 ccccccggcg acgcttcctg ctccgcccct aagtcgggaa ggttccttgc ggttcgcggc
1141 gtgccggacg tgacaaacgg aagccgcacg tctcactagt accctcgcag acggacagcg
1201 ccagggagca atggcagcgc gccgaccgcg atgggctgtg gccaatagcg gctgctcagc
1261 agggcgcgcc gagagcagcg gccgggaagg ggcggtgcgg gaggcggggt gtggggcggt
1321 agtgtgggcc ctgttcctgc ccgcgcggtg ttccgcattc tgcaagcctc cggagcgcac
1381 gtcggcagtc ggctccctcg ttgaccgaat caccgacctc tctccccagg gggtaccacc
1441 atggccaagc ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc
1501 aacagcatcc ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc
1561 cgcatcttca ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg
1621 gtgctgggca ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga
1681 aatgagaaca ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg
1741 catcctggga tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt
1801 cgtgaattgc tgccctctgg ttatgtgtgg gagggcctgc agctgcagta gtaaagacgc
1861 gttaagtcga caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
1921 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
1981 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
2041 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
2101 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
2161 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
2221 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
2281 cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
2341 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
2401 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
2461 gtcgacttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
2521 aaagggggga ctggaagggc taattcactc ccaacgaaga caagatctgc tttttgcttg
2581 tactgggtct ctctggttag accagatctg agcctgggag ctctctggct aactagggaa
2641 cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct
2701 gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc
2761 tagcagtacg tatagtagtt catgtcatct tattattcag tatttataac ttgcaaagaa
2821 atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt acaaataaag
2881 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
2941 gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc cctaactccg
3001 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
3061 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
3121 ggaggctttt ttggaggcct agggacgtac ccaattcgcc ctatagtgag tcgtattacg
3181 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
3241 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
3301 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg
3361 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
3421 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
3481 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
3541 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
3601 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
3661 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
3721 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
3781 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
3841 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
3901 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
3961 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
4021 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
4081 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
4141 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
4201 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
4261 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
4321 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
4381 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
4441 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
4501 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
4561 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
4621 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
4681 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
4741 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
4801 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
4861 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
4921 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
4981 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
5041 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
5101 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
5161 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
5221 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
5281 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
5341 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
5401 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
5461 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
5521 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
5581 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
5641 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
5701 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
5761 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
5821 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
5881 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
5941 aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa gggaacaaaa
6001 gctggagctg caagcttaat gtagtcttat gcaatactct tgtagtcttg caacatggta
6061 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
6121 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
6181 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
6241 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
6301 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
6361 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
6421 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
6481 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
6541 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
6601 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
6661 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
6721 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
6781 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
6841 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
6901 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cgctgatctt cagacctgga
6961 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
7021 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
7081 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
7141 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
7201 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
7261 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
7321 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
7381 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
7441 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
7501 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
7561 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
7621 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
7681 tattcaccat tatcgtttca gacccacctc ccaaccccga gggcgagctc ttacgcgtgc
7741 taggctgcaa gtcctgactt gtccacactc tgcaagtcct gacttgtccc taggctgcaa
7801 gtcctgactt gtccacactc tgcaagtcct gacttgtccc tagcccgggc tcgacagctg
7861 gacgtcgata tcgaattcgg gtatataatg gatccggtat cgagatctgc gatctaagta
7921 agcttggcat tccggtactg ttggtaaagc caccgaccc
//
