LOCUS Exported 6530 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses a fusion of full length CRY2 (E490G) with mCherry. ACCESSION . VERSION . KEYWORDS CRY2olig(FL)-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6530) AUTHORS Taslimi A, Vrana JD, Chen D, Borinskaya S, Mayer BJ, Kennedy MJ, Tucker CL TITLE An optimized optogenetic clustering tool for probing protein interaction and function. JOURNAL Nat Commun. 2014 Sep 18;5:4925. doi: 10.1038/ncomms5925. PUBMED 25233328 REFERENCE 2 (bases 1 to 6530) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6530 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 61..364 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 365..568 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 519..539 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 625..2460 /codon_start=1 /product="Arabidopsis thaliana cryptochrome 2 in which the nuclear localization signal was removed by mutating residues 541 and 542 to alanine" /label=CRY2(Delta-NLS) /note="binds to CIB1 upon exposure to blue light (Kennedy et al., 2010)" /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA SGVELGTNYAKPIVDIDTARELLAKAISRTRGAQIMIGAAPDEIVADSFEALGANTIKE PGLCPSVSSNDQQVPSAVRYNGSAAVKPEEEEERDMKKSRGFDERELFSTAESSSSSSV FFVSQSCSLASEGKNLEGIQDSSDQITTSLGKNGCK" regulatory 2479..2488 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2485..3195 /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(2630..2648) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2898..2917 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 3318..3439 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3355..3374) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3409..3428 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3446..3901) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3583..3604) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3795..3814 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3928..4032 /gene="bla" /label=AmpR promoter promoter 4034..4391 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4057..4077) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4242..4377 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4304..4323 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4426..5220 /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(4480..4499) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5090..5109 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5408..5427) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5452..5499 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5828..6416 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6317..6336 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg 61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta 601 ccggactcag atctcgaggc caccatgaag atggacaaaa agactatagt ttggtttaga 661 agagacctaa ggattgagga taatcctgca ttagcagcag ctgctcacga aggatctgtt 721 tttcctgtct tcatttggtg tcctgaagaa gaaggacagt tttatcctgg aagagcttca 781 agatggtgga tgaaacaatc acttgctcac ttatctcaat ccttgaaggc tcttggatct 841 gacctcactt taatcaaaac ccacaacacg atttcagcga tcttggattg tatccgcgtt 901 accggtgcta caaaagtcgt ctttaaccac ctctatgatc ctgtttcgtt agttcgggac 961 cataccgtaa aggagaagct ggtggaacgt gggatctctg tgcaaagcta caatggagat 1021 ctattgtatg aaccgtggga gatatactgc gaaaagggca aaccttttac gagtttcaat 1081 tcttactgga agaaatgctt agatatgtcg attgaatccg ttatgcttcc tcctccttgg 1141 cggttgatgc caataactgc agcggctgaa gcgatttggg cgtgttcgat tgaagaacta 1201 gggctggaga atgaggccga gaaaccgagc aatgcgttgt taactagagc ttggtctcca 1261 ggatggagca atgctgataa gttactaaat gagttcatcg agaagcagtt gatagattat 1321 gcaaagaaca gcaagaaagt tgttgggaat tctacttcac tactttctcc gtatctccat 1381 ttcggggaaa taagcgtcag acacgttttc cagtgtgccc ggatgaaaca aattatatgg 1441 gcaagagata agaacagtga aggagaagaa agtgcagatc tttttcttag gggaatcggt 1501 ttaagagagt attctcggta tatatgtttc aacttcccgt ttactcacga gcaatcgttg 1561 ttgagtcatc ttcggttttt cccttgggat gctgatgttg ataagttcaa ggcctggaga 1621 caaggcagga ccggttatcc gttggtggat gccggaatga gagagctttg ggctaccgga 1681 tggatgcata acagaataag agtgattgtt tcaagctttg ctgtgaagtt tcttctcctt 1741 ccatggaaat ggggaatgaa gtatttctgg gatacacttt tggatgctga tttggaatgt 1801 gacatccttg gctggcagta tatctctggg agtatccccg atggccacga gcttgatcgc 1861 ttggacaatc ccgcgttaca aggcgccaaa tatgacccag aaggtgagta cataaggcaa 1921 tggcttcccg agcttgcgag attgccaact gaatggatcc atcatccatg ggacgctcct 1981 ttaaccgtac tcaaagcttc tggtgtggaa ctcggaacaa actatgcgaa acccattgta 2041 gacatcgaca cagctcgtga gctactagct aaagctattt caagaacccg tggagcacag 2101 atcatgatcg gagcagcacc tgatgagatt gtagcagata gcttcgaggc cttaggggct 2161 aataccatta aagaacctgg tctttgccca tctgtgtctt ctaatgacca acaagtacct 2221 tcggctgttc gttacaacgg gtcagcggca gtgaaacctg aggaagaaga agagagagac 2281 atgaagaaat ctaggggatt cgatgaaagg gagttgtttt cgactgctga atcttcttct 2341 tcttcgagtg tgtttttcgt ttcgcagtct tgctcgttgg catcagaagg gaagaatctg 2401 gaaggtattc aagattcatc tgatcagatt actacaagtt tgggaaaaaa tggttgcaaa 2461 gcccgggatc caccggtcgc caccatggtg agcaagggcg aggaggataa catggccatc 2521 atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc 2581 gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc caagctgaag 2641 gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca gttcatgtac 2701 ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa gctgtccttc 2761 cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg 2821 acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct gcgcggcacc 2881 aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga ggcctcctcc 2941 gagcggatgt accccgagga cggcgccctg aagggcgaga tcaagcagag gctgaagctg 3001 aaggacggcg gccactacga cgctgaggtc aagaccacct acaaggccaa gaagcccgtg 3061 cagctgcccg gcgcctacaa cgtcaacatc aagttggaca tcacctccca caacgaggac 3121 tacaccatcg tggaacagta cgaacgcgcc gagggccgcc actccaccgg cggcatggac 3181 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag 3241 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga 3301 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata 3361 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca 3421 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg 3481 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct 3541 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt 3601 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat 3661 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca 3721 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac 3781 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta 3841 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg 3901 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata 3961 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga 4021 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg 4081 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 4141 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 4201 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 4261 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 4321 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 4381 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat 4441 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca 4501 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg 4561 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg 4621 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact 4681 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct 4741 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg 4801 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt 4861 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc 4921 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc 4981 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga 5041 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc 5101 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt 5161 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga 5221 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt 5281 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg 5341 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga 5401 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa 5461 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc 5521 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt 5581 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg 5641 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa 5701 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca 5761 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag 5821 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac 5881 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa 5941 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc 6001 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag 6061 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac 6121 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc 6181 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc 6241 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca 6301 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc 6361 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg 6421 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct 6481 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat //