LOCUS Exported 6188 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses CRY2(1-498)-mCherry fusion for use in light-inducible protein interaction modules. ACCESSION . VERSION . KEYWORDS pCRY2PHR-mCherryN1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6188) AUTHORS Kennedy MJ, Hughes RM, Peteya LA, Schwartz JW, Ehlers MD, Tucker CL TITLE Rapid blue-light-mediated induction of protein interactions in living cells. JOURNAL Nat Methods. 2010 Oct 31. ():. PUBMED 21037589 REFERENCE 2 (bases 1 to 6188) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6188 /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..2118 /codon_start=1 /product="N-terminal photolyase homology region of Arabidopsis thaliana cryptochrome 2" /label=CRY2PHR /note="binds to CIB1 upon exposure to blue light (Kennedy et al., 2010)" /translation="MKMDKKTIVWFRRDLRIEDNPALAAAAHEGSVFPVFIWCPEEEGQ FYPGRASRWWMKQSLAHLSQSLKALGSDLTLIKTHNTISAILDCIRVTGATKVVFNHLY DPVSLVRDHTVKEKLVERGISVQSYNGDLLYEPWEIYCEKGKPFTSFNSYWKKCLDMSI ESVMLPPPWRLMPITAAAEAIWACSIEELGLENEAEKPSNALLTRAWSPGWSNADKLLN EFIEKQLIDYAKNSKKVVGNSTSLLSPYLHFGEISVRHVFQCARMKQIIWARDKNSEGE ESADLFLRGIGLREYSRYICFNFPFTHEQSLLSHLRFFPWDADVDKFKAWRQGRTGYPL VDAGMRELWATGWMHNRIRVIVSSFAVKFLLLPWKWGMKYFWDTLLDADLECDILGWQY ISGSIPDGHELDRLDNPALQGAKYDPEGEYIRQWLPELARLPTEWIHHPWDAPLTVLKA SGVELGTNYAKPIVDIDTARELLAKAISRTREAQIMIGAA" regulatory 2137..2146 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2143..2853 /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(2288..2306) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2556..2575 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2976..3097 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3013..3032) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3067..3086 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3104..3559) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3241..3262) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3453..3472 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3586..3690 /gene="bla" /label=AmpR promoter promoter 3692..4049 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3715..3735) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3900..4035 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3962..3981 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4084..4878 /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(4138..4157) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4748..4767 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5066..5085) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5110..5157 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5486..6074 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5975..5994 /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 tgaagcacag 2101 atcatgatcg gagcagcagc ccgggatcca ccggtcgcca ccatggtgag caagggcgag 2161 gaggataaca tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc 2221 gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc 2281 cagaccgcca agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg 2341 tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac 2401 tacttgaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac 2461 ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag 2521 gtgaagctgc gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaagaccatg 2581 ggctgggagg cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc 2641 aagcagaggc tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac 2701 aaggccaaga agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc 2761 acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac 2821 tccaccggcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca 2881 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga 2941 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg 3001 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt 3061 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat 3121 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc 3181 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt 3241 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa 3301 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg 3361 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga 3421 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct 3481 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat 3541 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt 3601 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa 3661 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg 3721 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3781 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 3841 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 3901 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 3961 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 4021 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt 4081 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta 4141 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg 4201 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa 4261 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct 4321 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg 4381 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca 4441 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat 4501 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac 4561 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc 4621 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa 4681 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag 4741 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc 4801 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt 4861 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca 4921 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa 4981 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct 5041 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc 5101 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata 5161 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg 5221 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc 5281 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata 5341 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 5401 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 5461 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 5521 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 5581 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 5641 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 5701 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 5761 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 5821 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 5881 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 5941 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 6001 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 6061 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 6121 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 6181 ccatgcat //