LOCUS Exported 6886 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses fusion of CRY2low mutant (CRY2 aa1-491 R489E A491D) with tdTomato. ACCESSION . VERSION . KEYWORDS CRY2low-tdTomato SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6886) AUTHORS Duan L, Hope J, Ong Q, Lou HY, Kim N, McCarthy C, Acero V, Lin MZ, Cui B TITLE Understanding CRY2 interactions for optical control of intracellular signaling. JOURNAL Nat Commun. 2017 Sep 15;8(1):547. doi: 10.1038/s41467-017-00648-8. PUBMED 28916751 REFERENCE 2 (bases 1 to 6886) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6886 /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" regulatory 2116..2125 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2122..3552 /codon_start=1 /product="tandem dimeric (pseudo-monomeric) derivative of DsRed (Shaner et al., 2004)" /label=tdTomato /note="mammalian codon-optimized" /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE RSEGRHHLFLYGMDELYK" polyA_signal 3674..3795 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3711..3730) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3765..3784 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(3802..4257) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3939..3960) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4151..4170 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4284..4388 /gene="bla" /label=AmpR promoter promoter 4390..4747 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4413..4433) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4598..4733 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4660..4679 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4782..5576 /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(4836..4855) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5446..5465 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5764..5783) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 5808..5855 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6184..6772 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6673..6692 /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 caagaaccga ggaagacgcc 2101 cgggataagg atcccgccac catggtgagc aagggcgagg aggtcatcaa agagttcatg 2161 cgcttcaagg tgcgcatgga gggctccatg aacggccacg agttcgagat cgagggcgag 2221 ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac caagggcggc 2281 cccctgccct tcgcctggga catcctgtcc ccccagttca tgtacggctc caaggcgtac 2341 gtgaagcacc ccgccgacat ccccgattac aagaagctgt ccttccccga gggcttcaag 2401 tgggagcgcg tgatgaactt cgaggacggc ggtctggtga ccgtgaccca ggactcctcc 2461 ctgcaggacg gcacgctgat ctacaaggtg aagatgcgcg gcaccaactt cccccccgac 2521 ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct ccaccgagcg cctgtacccc 2581 cgcgacggcg tgctgaaggg cgagatccac caggccctga agctgaagga cggcggccac 2641 tacctggtgg agttcaagac catctacatg gccaagaagc ccgtgcaact gcccggctac 2701 tactacgtgg acaccaagct ggacatcacc tcccacaacg aggactacac catcgtggaa 2761 cagtacgagc gctccgaggg ccgccaccac ctgttcctgg ggcatggcac cggcagcacc 2821 ggcagcggca gctccggcac cgcctcctcc gaggacaaca acatggccgt catcaaagag 2881 ttcatgcgct tcaaggtgcg catggagggc tccatgaacg gccacgagtt cgagatcgag 2941 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag 3001 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttcatgta cggctccaag 3061 gcgtacgtga agcaccccgc cgacatcccc gattacaaga agctgtcctt ccccgagggc 3121 ttcaagtggg agcgcgtgat gaacttcgag gacggcggtc tggtgaccgt gacccaggac 3181 tcctccctgc aggacggcac gctgatctac aaggtgaaga tgcgcggcac caacttcccc 3241 cccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg 3301 tacccccgcg acggcgtgct gaagggcgag atccaccagg ccctgaagct gaaggacggc 3361 ggccactacc tggtggagtt caagaccatc tacatggcca agaagcccgt gcaactgccc 3421 ggctactact acgtggacac caagctggac atcacctccc acaacgagga ctacaccatc 3481 gtggaacagt acgagcgctc cgagggccgc caccacctgt tcctgtacgg catggacgag 3541 ctgtacaagt aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt 3601 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc 3661 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 3721 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 3781 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa 3841 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata 3901 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac 3961 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc 4021 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa 4081 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg 4141 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg 4201 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag 4261 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 4321 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 4381 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa 4441 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 4501 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 4561 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 4621 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 4681 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt 4741 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 4801 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 4861 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 4921 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc 4981 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 5041 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 5101 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 5161 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 5221 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 5281 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga 5341 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 5401 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 5461 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 5521 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 5581 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 5641 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 5701 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct 5761 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag 5821 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg 5881 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc 5941 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg 6001 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact 6061 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 6121 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 6181 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 6241 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 6301 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca 6361 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 6421 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 6481 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 6541 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 6601 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 6661 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 6721 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 6781 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 6841 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat //