LOCUS Exported 5240 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of light-regulated JNK inhibitor, mCherry-tagged, localised to nucleus, lit-state mutant photosensor (AsLov2Ja.I539E), inhibitor JIP11. ACCESSION . VERSION . KEYWORDS pmCherry-3xNLS-OptoJNKi-lsm SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5240) AUTHORS Melero-Fernandez de Mera RM, Li LL, Popinigis A, Cisek K, Tuittila M, Yadav L, Serva A, Courtney MJ TITLE A simple optogenetic MAPK inhibitor design reveals resonance between transcription-regulating circuitry and temporally-encoded inputs. JOURNAL Nat Commun. 2017 May 12;8:15017. doi: 10.1038/ncomms15017. PUBMED 28497795 REFERENCE 2 (bases 1 to 5240) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5240 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 144..447 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 448..651 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 602..622 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 690..699 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 696..1403 /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(841..859) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1109..1128 /label=mCherry-F /note="mCherry, forward primer" CDS 1446..1466 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 2111..2232 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2148..2167) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2202..2221 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2239..2694) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2376..2397) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2588..2607 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2721..2825 /gene="bla" /label=AmpR promoter promoter 2827..3184 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2850..2870) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3035..3170 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3097..3116 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3219..4013 /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 GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3273..3292) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3883..3902 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4201..4220) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4245..4292 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4621..5209 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5110..5129 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg 61 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt 121 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga 181 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca 241 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca 301 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg 361 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc 421 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt 481 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt 541 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg 601 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg 661 aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggaggata 721 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg 781 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg 841 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc 901 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga 961 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg 1021 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc 1081 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg 1141 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga 1201 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca 1261 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc 1321 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg 1381 gcggcatgga cgagctgtac aagtccggac tcagatccga ccccagaaga aagcgcaagg 1441 tggaccccaa gaagaagcgc aaggtggacc ccaagaagaa gcgcagatct cgagctcaag 1501 cttcgaattc actggcaaca acactggaga gaatcgagaa aaacttcgtc atcaccgacc 1561 ctagactgcc cgataatcct atcatcttcg cctccgactc atttctccag ctcactgagt 1621 actctcggga ggaaattctg ggccggaatt gccggttcct ccagggacct gaaaccgata 1681 gagccactgt ccgaaaaatc cgggatgcca tcgacaatca gactgaggtg accgtccagc 1741 tcatcaatta caccaaatct ggcaaaaaat tctggaacct gttccacctc cagcctatga 1801 gagatcagaa gggcgacgtc cagtacttta ttggcgtcca gctggatgga actgaacatg 1861 tgagggacgc tgcagaacga gagggcgtga tgctcatcaa aaaaaccgcc gaaaacgagg 1921 acgaggccgc taaagagctg tctagaagac ccaaacgacc aaccacactg aacctcttct 1981 aggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt 2041 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat 2101 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac 2161 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat 2221 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt 2281 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat 2341 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat 2401 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac 2461 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc 2521 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga 2581 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca 2641 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg 2701 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa 2761 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga 2821 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc 2881 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag 2941 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 3001 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc 3061 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc 3121 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg 3181 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc 3241 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga 3301 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt 3361 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat 3421 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg 3481 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg 3541 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc 3601 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga 3661 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag 3721 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc 3781 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg 3841 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata 3901 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg 3961 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac 4021 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc 4081 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 4141 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac 4201 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca 4261 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg 4321 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt 4381 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc 4441 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca 4501 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 4561 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 4621 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 4681 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 4741 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 4801 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 4861 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 4921 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 4981 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 5041 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 5101 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 5161 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 5221 cgcggccttt ttacggttcc //