| 质粒名称: | EK0208 CMV-TO-BFPmut-stop-t70587 (FLP-IN) |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 191138 |
| 存储实验室: | Xiaojing Gao |
|
载体骨架基本信息 |
|
| 载体名称: | FLP-IN |
| 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: | t70587 |
| 物种来源: | Synthetic |
| 基因突变: | WT |
| 启动子: | CMV-TO |
| 克隆方法: | 未知 |
|
文献引用方法 |
|
| 材料和方法部分: | EK0208 CMV-TO-BFPmut-stop-t70587 (FLP-IN) was a gift from Xiaojing Gao (Addgene plasmid # 191138 ; http://n2t.net/addgene:191138 ; |
| 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
质粒图谱.png)
载体简介
载体序列
LOCUS EK0208 CMV-TO-BFPmut-stop-t70587 (FLP-IN) 6022 bp ds-DNA circular SYN 25-OCT-2022
DEFINITION Expression of Trigger 1 in 3' UTR of an inactive BFP in mammalian
cells.
ACCESSION .
VERSION .
KEYWORDS EK0208 CMV-TO-BFPmut-stop-t70587 (FLP-IN)
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6022)
AUTHORS Kaseniit E, Gao X
TITLE RADAR plasmids
JOURNAL unpublished
REFERENCE 2 (bases 1 to 6022)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Oct 25, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6022
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 4..383
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 384..587
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 538..558
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
protein_bind 589..607
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 610..628
/gene="tetO"
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 638..662
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 754..763
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 769..1476
/codon_start=1
/product="enhanced monomeric blue fluorescent protein
(Subach et al., 2011)"
/label=mTagBFP2
/note="mammalian codon-optimized"
/translation="VSKGEELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMR
IKVVEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
LTATQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDM
ALKLVGGSHLIANAKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHAVAVA
RYCDLPSKLGHKLN"
CDS 1495..1521
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1624..1653
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
primer_bind complement(1709..1726)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 1715..1939
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(2111..2130)
/label=F1ori-R
/note="F1 origin, reverse primer"
protein_bind 2262..2309
/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 2317..3339
/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 3469..3590
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(3506..3525)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 3560..3579
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(3639..3655)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3639..3655)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3652..3674)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3663..3679
/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(3687..3717)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3732..3753
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3870..3887)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4041..4629)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4121..4140)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4800..5660)
/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 5423..5442
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5661..5765)
/gene="bla"
/label=AmpR promoter
primer_bind complement(5840..5859)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc cctatcagtg
601 atagagatct ccctatcagt gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga
661 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
721 gcctccggac tctagcgttt aaacttaagc ttcgccacca tgggatccgt gtctaagggc
781 gaagagctga ttaaggagaa catgcacatg aagctgtaca tggagggcac cgtggacaac
841 catcacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac ccagaccatg
901 agaatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct ggctactagc
961 ttcctctacg gcagcaagac cttcatcaac cacacccagg gcatccccga cttcttcaag
1021 cagtccttcc ctgagggctt cacatgggag agagtcacca catacgagga cgggggcgtg
1081 ctgaccgcta cccaggacac cagcctccag gacggctgcc tcatctacaa cgtcaagatc
1141 agaggggtga acttcacatc caacggccct gtgatgcaga agaaaacact cggctgggag
1201 gccttcaccg aaacgctgta ccccgctgac ggcggcctgg aaggcagaaa cgacatggcc
1261 ctgaagctcg tgggcgggag ccatctgatc gcaaacgcca agaccacata tagatccaag
1321 aaacccgcta agaacctcaa gatgcctggc gtctactatg tggactacag actggaaaga
1381 atcaaggagg ccaacaacga aacctacgtc gagcagcacg cggtggcagt ggccagatac
1441 tgcgacctcc ctagcaaact ggggcacaaa cttaattgaa gctagctggc tggataccca
1501 tacgatgttc cagattacgc tctcgagata gctctctatt tgcaccagtg gtctatcggt
1561 acgtaaggcc caacaatgac gcggatagaa ggaaacactc atccgtgtcg acttggtggg
1621 cccgaacaaa aactcatctc agaagaggat ctgaatagcg ccgtcgacca tcatcataat
1681 catcattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
1741 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
1801 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
1861 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
1921 gatgcggtgg gctctatgga gatctatcga tgttctgggg atgcggtggg ctctatggct
1981 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
2041 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
2101 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
2161 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
2221 gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt
2281 tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg
2341 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
2401 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
2461 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
2521 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
2581 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
2641 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
2701 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
2761 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
2821 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
2881 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
2941 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
3001 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
3061 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
3121 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
3181 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
3241 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
3301 aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg agatttcgat
3361 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
3421 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
3481 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
3541 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
3601 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
3661 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
3721 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
3781 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
3841 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
3901 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
3961 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
4021 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
4081 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
4141 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
4201 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
4261 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
4321 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
4381 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
4441 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
4501 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
4561 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
4621 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
4681 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
4741 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
4801 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
4861 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
4921 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
4981 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
5041 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
5101 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
5161 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
5221 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
5281 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
5341 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
5401 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
5461 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
5521 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
5581 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
5641 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
5701 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
5761 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
5821 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
5881 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
5941 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
6001 ttcgtacggg ccagatatac gc
//
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