| 质粒名称: | EK0294 SFFV-mCherry-SG(s70587-W)SG-EGFP (FLP-IN) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 191141 |
| 存储实验室: | Xiaojing Gao |
|
载体骨架基本信息 |
|
| 载体名称: | FLP-IN |
| 完整质粒大小: | 6590bp |
| 载体类型: | 哺乳细胞表达 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Cherry:s70587-W:EGFP |
| 物种来源: | Synthetic |
| 基因突变: | UAG>UGG (stop>Trp) |
| 启动子: | SFFV |
| 克隆方法: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | EK0294 SFFV-mCherry-SG(s70587-W)SG-EGFP (FLP-IN) was a gift from Xiaojing Gao (Addgene plasmid # 191141 ; http://n2t.net/addgene:191141 ; RRID:Addgene_191141) |
| 参考文献部分: | Modular, programmable RNA sensing using ADAR editing in living cells. Kaseniit KE, Katz N, Kolber NS, Call CC, Wengier DL, Cody WB, Sattely ES, Gao XJ. Nat Biotechnol. 2022 Oct 5. pii: 10.1038/s41587-022-01493-x. doi: 10.1038/s41587-022-01493-x. 10.1038/s41587-022-01493-x PubMed 36198772 |
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
SG-EGFP (FLP-IN)质粒图谱.png)
载体简介
载体序列
LOCUS EK0294 SFFV-mCherry-SG(s70587-W)SG-EGFP (FLP-IN) 6590 bp ds-DNA circular SYN 26-OCT-2022
DEFINITION Positive control for Sensor 1 (EK0285) where the TAG stop codon is
replaced with TGG tryptophan codon.
ACCESSION .
VERSION .
KEYWORDS EK0294 SFFV-mCherry-SG(s70587-W)SG-EGFP (FLP-IN)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6590)
AUTHORS Kaseniit E, Gao X
TITLE RADAR plasmids
JOURNAL unpublished
REFERENCE 2 (bases 1 to 6590)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 26, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6590
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 90..497
/label=SFFV promoter
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
regulatory 505..514
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 511..1218
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(656..674)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 924..943
/label=mCherry-F
/note="mCherry, forward primer"
CDS 1255..1320
/codon_start=1
/product="2A peptide from foot-and-mouth disease virus
polyprotein"
/label=F2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="VKQTLNFDLLKLAGDVESNPGP"
CDS 1456..1521
/codon_start=1
/product="2A peptide from foot-and-mouth disease virus
polyprotein"
/label=F2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="VKQTLNFDLLKLAGDVESNPGP"
CDS 1522..2235
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
CDS 2257..2286
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(2342..2359)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2348..2572
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(2744..2763)
/label=F1ori-R
/note="F1 origin, reverse primer"
protein_bind 2895..2942
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 2950..3972
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
"
polyA_signal 4102..4223
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4139..4158)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4193..4212
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4272..4288)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4272..4288)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4285..4307)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4296..4312
/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(4320..4350)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4365..4386
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4503..4520)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4674..5262)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4754..4773)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5433..6293)
/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 6056..6075
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6294..6398)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6473..6492)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 aattgccgat aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc
61 ccacctgtag gtttggcaag ctagctgcag taacgccatt ttgcaaggca tggaaaaata
121 ccaaaccaag aatagagaag ttcagatcaa gggcgggtac atgaaaatag ctaacgttgg
181 gccaaacagg atatctgcgg tgagcagttt cggccccggc ccggggccaa gaacagatgg
241 tcaccgcagt ttcggccccg gcccgaggcc aagaacagat ggtccccaga tatggcccaa
301 ccctcagcag tttcttaaga cccatcagat gtttccaggc tcccccaagg acctgaaatg
361 accctgcgcc ttatttgaat taaccaatca gcctgcttct cgcttctgtt cgcgcgcttc
421 tgcttcccga gctctataaa agagctcaca acccctcact cggcgcgcca gtcctccgac
481 agactgagtc gcccgggaag cttcgccacc atggtgagca agggcgagga ggataacatg
541 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
601 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
661 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
721 atgtacggct ccaaggccta cgtgaagcac cccgccgaca ttcccgacta cttgaagctg
781 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
841 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
901 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
961 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
1021 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
1081 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
1141 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
1201 atggacgagc tgtacaagag cgaagccaga cacaaacaga aaattgtggc tccggtgaaa
1261 cagactttga attttgacct tctgaagctg gcgggagacg tcgagtccaa ccccggtccc
1321 ggatccacca agtcgacacg gatgagtgtt tccttctatc cgcgtcattg ttgggcctta
1381 cgtaccgata gaccactggt gcaaatagag agctataccg gtgaggcacg gcataagcaa
1441 aagatcgtag ccccagtaaa gcaaacactc aacttcgatc tactcaaact cgcaggtgat
1501 gtggaatcta atccaggacc tgtctcaaag ggcgaggaac tttttacggg cgttgttccg
1561 atcctcgtcg aattggacgg ggacgttaat gggcataaat tctcagttag tggtgagggt
1621 gagggggacg cgacatatgg gaaacttacg ctgaagttca tatgtaccac cggaaaattg
1681 ccagtgccgt ggcctactct cgtcaccacc ctgacatatg gtgttcagtg ttttagccgg
1741 tatccagacc atatgaaaca gcacgatttt tttaaatccg ctatgcctga aggttacgta
1801 caggagcgaa cgatattttt taaagacgac ggaaactaca aaactagagc cgaggtaaaa
1861 ttcgagggtg acactctcgt gaaccggatc gaactgaagg gcatagattt taaggaggat
1921 ggcaatatac tgggccacaa gctcgaatac aactataatt ctcataatgt ctatatcatg
1981 gcggacaaac agaaaaacgg catcaaggta aactttaaaa tccgacacaa tattgaggac
2041 ggctctgtcc agcttgcgga ccattaccaa cagaacacgc cgatagggga tggtccagta
2101 cttctccctg ataatcacta tttgtctact cagtcagctc ttagcaagga cccaaacgaa
2161 aaacgggatc acatggtcct tcttgagttc gtcactgcgg ctgggataac actcgggatg
2221 gacgaactgt ataagtaggc cgctcgagga gggcccgaac aaaaactcat ctcagaagag
2281 gatctgaata gcgccgtcga ccatcatcat aatcatcatt gagtttaaac ccgctgatca
2341 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
2401 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
2461 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
2521 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggagatctat
2581 cgatgttctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
2641 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
2701 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
2761 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
2821 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
2881 ggttcacgta cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact
2941 tccttggcca aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag
3001 ttcgacagcg tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc
3061 ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac
3121 aaagatcgtt atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt
3181 gacattgggg aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc
3241 acgttgcaag acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc
3301 atggatgcga tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg
3361 caaggaatcg gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat
3421 gtgtatcact ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc
3481 gatgagctga tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat
3541 ttcggctcca acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc
3601 gaggcgatgt tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg
3661 ttggcttgta tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga
3721 tcgccgcggc tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg
3781 gttgacggca atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga
3841 tccggagccg ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc
3901 gatggctgtg tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg
3961 gcaaaggaat agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg
4021 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
4081 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
4141 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
4201 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
4261 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
4321 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
4381 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
4441 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
4501 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
4561 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
4621 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
4681 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
4741 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
4801 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
4861 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
4921 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
4981 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
5041 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
5101 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
5161 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
5221 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
5281 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
5341 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
5401 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
5461 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
5521 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
5581 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
5641 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
5701 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
5761 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
5821 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
5881 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
5941 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
6001 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
6061 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
6121 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
6181 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
6241 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
6301 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
6361 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
6421 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
6481 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
6541 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
//
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