| 质粒名称: | EK0285 SFFV-mCherry-SG(s70587)SG-EGFP (FLP-IN) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 191140 |
| 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: | mCherry:s70587:EGFP |
| 物种来源: | Synthetic |
| 基因突变: | WT |
| 启动子: | SFFV |
| 克隆方法: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | EK0285 SFFV-mCherry-SG(s70587)SG-EGFP (FLP-IN) was a gift from Xiaojing Gao (Addgene plasmid # 191140 ; http://n2t.net/addgene:191140 ; RRID:Addgene_191140) |
| 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 EK0285 SFFV-mCherry-SG(s70587)SG-EGFP (FLP-IN) 6590 bp ds-DNA circular SYN 25-OCT-2022
DEFINITION Expression of Sensor 1 (for EK0208) in mammalian cells; mCherry
marker, EGFP output..
ACCESSION .
VERSION .
KEYWORDS EK0285 SFFV-mCherry-SG(s70587)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JOURNAL unpublished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 25, 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 176..583
/label=SFFV promoter
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
regulatory 591..600
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 597..1304
/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(742..760)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 1010..1029
/label=mCherry-F
/note="mCherry, forward primer"
CDS 1341..1406
/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 1542..1607
/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 1608..2321
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
CDS 2343..2372
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(2428..2445)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2434..2658
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(2830..2849)
/label=F1ori-R
/note="F1 origin, reverse primer"
protein_bind 2981..3028
/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 3036..4058
/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 4188..4309
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4225..4244)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4279..4298
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4358..4374)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4358..4374)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4371..4393)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4382..4398
/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(4406..4436)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4451..4472
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4589..4606)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4760..5348)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4840..4859)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5519..6379)
/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 6142..6161
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6380..6484)
/gene="bla"
/label=AmpR promoter
primer_bind complement(6559..6578)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
61 ctacaacaag gcaaggcttg accgacaatt gccgataaaa taaaagattt tatttagtct
121 ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag ctgcagtaac
181 gccattttgc aaggcatgga aaaataccaa accaagaata gagaagttca gatcaagggc
241 gggtacatga aaatagctaa cgttgggcca aacaggatat ctgcggtgag cagtttcggc
301 cccggcccgg ggccaagaac agatggtcac cgcagtttcg gccccggccc gaggccaaga
361 acagatggtc cccagatatg gcccaaccct cagcagtttc ttaagaccca tcagatgttt
421 ccaggctccc ccaaggacct gaaatgaccc tgcgccttat ttgaattaac caatcagcct
481 gcttctcgct tctgttcgcg cgcttctgct tcccgagctc tataaaagag ctcacaaccc
541 ctcactcggc gcgccagtcc tccgacagac tgagtcgccc gggaagcttc gccaccatgg
601 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
661 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
721 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
781 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
841 ccgacattcc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
901 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
961 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
1021 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
1081 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
1141 tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
1201 tcaagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg
1261 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagagcgaa gccagacaca
1321 aacagaaaat tgtggctccg gtgaaacaga ctttgaattt tgaccttctg aagctggcgg
1381 gagacgtcga gtccaacccc ggtcccggat ccaccaagtc gacacggatg agtgtttcct
1441 tctatccgcg tcattgttag gccttacgta ccgatagacc actggtgcaa atagagagct
1501 ataccggtga ggcacggcat aagcaaaaga tcgtagcccc agtaaagcaa acactcaact
1561 tcgatctact caaactcgca ggtgatgtgg aatctaatcc aggacctgtc tcaaagggcg
1621 aggaactttt tacgggcgtt gttccgatcc tcgtcgaatt ggacggggac gttaatgggc
1681 ataaattctc agttagtggt gagggtgagg gggacgcgac atatgggaaa cttacgctga
1741 agttcatatg taccaccgga aaattgccag tgccgtggcc tactctcgtc accaccctga
1801 catatggtgt tcagtgtttt agccggtatc cagaccatat gaaacagcac gattttttta
1861 aatccgctat gcctgaaggt tacgtacagg agcgaacgat attttttaaa gacgacggaa
1921 actacaaaac tagagccgag gtaaaattcg agggtgacac tctcgtgaac cggatcgaac
1981 tgaagggcat agattttaag gaggatggca atatactggg ccacaagctc gaatacaact
2041 ataattctca taatgtctat atcatggcgg acaaacagaa aaacggcatc aaggtaaact
2101 ttaaaatccg acacaatatt gaggacggct ctgtccagct tgcggaccat taccaacaga
2161 acacgccgat aggggatggt ccagtacttc tccctgataa tcactatttg tctactcagt
2221 cagctcttag caaggaccca aacgaaaaac gggatcacat ggtccttctt gagttcgtca
2281 ctgcggctgg gataacactc gggatggacg aactgtataa gtaggccgct cgaggagggc
2341 ccgaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat catcataatc
2401 atcattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
2461 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
2521 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
2581 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
2641 atgcggtggg ctctatggag atctatcgat gttctgggga tgcggtgggc tctatggctt
2701 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
2761 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
2821 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
2881 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
2941 accccaaaaa acttgattag ggtgatggtt cacgtaccta gaagttccta ttccgaagtt
3001 cctattctct agaaagtata ggaacttcct tggccaaaaa gcctgaactc accgcgacgt
3061 ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg cagctctcgg
3121 agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat gtcctgcggg
3181 taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac tttgcatcgg
3241 ccgcgctccc gattccggaa gtgcttgaca ttggggaatt cagcgagagc ctgacctatt
3301 gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc gaactgcccg
3361 ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat cttagccaga
3421 cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca tggcgtgatt
3481 tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg gacgacaccg
3541 tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag gactgccccg
3601 aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg gacaatggcc
3661 gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa tacgaggtcg
3721 ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg cgctacttcg
3781 agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg ctccgcattg
3841 gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca gcttgggcgc
3901 agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt acacaaatcg
3961 cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc gatagtggaa
4021 accgacgccc cagcactcgt ccgagggcaa aggaatagca cgtactacga gatttcgatt
4081 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
4141 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg
4201 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
4261 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta
4321 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
4381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
4441 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
4501 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
4561 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
4621 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
4681 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
4741 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
4801 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
4861 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
4921 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
4981 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
5041 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
5101 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
5161 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
5221 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
5281 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
5341 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
5401 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
5461 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
5521 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
5581 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
5641 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
5701 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
5761 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
5821 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
5881 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
5941 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
6001 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
6061 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
6121 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
6181 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
6241 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
6301 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
6361 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
6421 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
6481 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc
6541 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
//
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