LOCUS Exported 5234 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expression of light-regulated JNK inhibitor, mCherry-tagged, localised to cytoplasm, wild-type photosensor, inhibitor JIP11. ACCESSION . VERSION . KEYWORDS pmCherry-NES-OptoJNKi SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5234) 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 5234) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5234 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 126..429 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 430..633 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 584..604 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 672..681 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 678..1385 /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(823..841) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1091..1110 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2087..2208 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2124..2143) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2178..2197 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2215..2670) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2352..2373) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2564..2583 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2697..2801 /gene="bla" /label=AmpR promoter promoter 2803..3160 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(2826..2846) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3011..3146 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3073..3092 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3195..3989 /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(3249..3268) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 3859..3878 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4177..4196) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4221..4268 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4597..5185 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5086..5105 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca 61 tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag 121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc 181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga 241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat 301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc 361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct 421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca 481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat 541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg 601 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag 661 cgctaccggt cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg 721 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg 781 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca 841 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca 901 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg 961 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg 1021 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc 1081 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga 1141 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg 1201 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc 1261 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca 1321 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt 1381 acaagtccgg actcagatcc gccctgcaga agaagctgga ggagctggag ctggacgagc 1441 agcagcgaaa gcgcctcgag agatctcgag ctcaagcttc gaattcactg gcaacaacac 1501 tggagagaat cgagaaaaac ttcgtcatca ccgaccctag actgcccgat aatcctatca 1561 tcttcgcctc cgactcattt ctccagctca ctgagtactc tcgggaggaa attctgggcc 1621 ggaattgccg gttcctccag ggacctgaaa ccgatagagc cactgtccga aaaatccggg 1681 atgccatcga caatcagact gaggtgaccg tccagctcat caattacacc aaatctggca 1741 aaaaattctg gaacctgttc cacctccagc ctatgagaga tcagaagggc gacgtccagt 1801 actttattgg cgtccagctg gatggaactg aacatgtgag ggacgctgca gaacgagagg 1861 gcgtgatgct catcaaaaaa accgccgaaa acatcgatga ggccgctaaa gagctgtcta 1921 gaagacccaa acgaccaacc acactgaacc tcttctagga tccaccggat ctagataact 1981 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca 2041 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc 2101 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt 2161 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtaaattg 2221 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta 2281 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt 2341 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca 2401 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa 2461 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat 2521 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag 2581 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg 2641 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg 2701 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 2761 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca 2821 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag 2881 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc 2941 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct 3001 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg 3061 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa 3121 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat 3181 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 3241 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 3301 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 3361 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 3421 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 3481 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 3541 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 3601 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 3661 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 3721 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 3781 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 3841 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 3901 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 3961 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 4021 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 4081 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 4141 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc 4201 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc 4261 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag 4321 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc 4381 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg 4441 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt 4501 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg 4561 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 4621 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 4681 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 4741 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 4801 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 4861 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 4921 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 4981 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 5041 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 5101 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 5161 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 5221 cttttgctgg cctt //