LOCUS Exported 6046 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses mCherry fused to UVR8 under the CMV promoter. ACCESSION . VERSION . KEYWORDS UVR8-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6046) AUTHORS Chen D, Gibson ES, Kennedy MJ TITLE A light-triggered protein secretion system. JOURNAL J Cell Biol. 2013 May 13;201(4):631-40. doi: 10.1083/jcb.201210119. PUBMED 23671313 REFERENCE 2 (bases 1 to 6046) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6046 /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 618..627 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 1995..2004 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2001..2711 /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(2146..2164) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2414..2433 /label=mCherry-F /note="mCherry, forward primer" polyA_signal 2834..2955 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2871..2890) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2925..2944 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2962..3417) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3099..3120) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3311..3330 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3444..3548 /gene="bla" /label=AmpR promoter promoter 3550..3907 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3573..3593) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3758..3893 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3820..3839 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3942..4736 /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(3996..4015) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4606..4625 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4924..4943) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4968..5015 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5344..5932 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5833..5852 /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 atctcgagcc accatggcgg aggatatggc tgccgacgaa gttacggctc 661 ctcctcgtaa ggttcttatc atctccgctg gtgctagcca ctccgtcgct cttctctctg 721 gtgacattgt ttgttcttgg ggtcgaggag aggatggaca gttaggtcat ggcgatgcag 781 aggatcgacc ttctccgact cagcttagcg ctttagatgg ccaccaaatt gtttccgtta 841 cctgtggtgc tgatcacact gttgcttatt cacaatcagg catggaagtc tacagttggg 901 gatggggtga ttttgggaga ttaggccatg gtaactcaag cgacttgttt actccgctac 961 caatcaaagc attgcacggt attcggatca agcagattgc ttgtggggat agtcattgtt 1021 tggctgtcac tatggaagga gaggtccaga gttggggccg caaccagaat ggtcaacttg 1081 gtctggggga caccgaagat tctctagtgc ctcagaagat tcaagccttt gagggaatac 1141 gaatcaaaat ggttgctgct ggtgcagaac acactgctgc agttacagaa gatggtgacc 1201 tctatggatg gggctgggga agatacggaa atttgggatt aggtgaccgg actgaccgct 1261 tagttcctga aagagttacc tctactggtg gtgagaaaat gtcaatggtt gcttgtggat 1321 ggcggcacac aatatcagtt tcctactctg gagcattgta tacttatgga tggagcaaat 1381 atggacagct aggacatgga gacttggagg atcaccttat tcctcacaaa ctggaagcac 1441 tgagcaacag ttttatctcc cagatttcgg gaggttggag acatacaatg gcattgactt 1501 cagatggaaa actatatgga tggggttgga ataagtttgg acaagtagga gtcggcaata 1561 atttagatca gtgttctcct gtgcaagtgc gatttcccga tgatcagaaa gtagttcaag 1621 tctcatgtgg atggagacat accttggctg tcactgaaag aaataacgtg tttgcttggg 1681 gtagaggtac aaatggacag ctcggcattg gagagtcggt tgacaggaac tttcccaaga 1741 ttatagaggc actcagcgtc gatggagcaa gtggacaaca tatagaatct tctaatatcg 1801 atccatcttc agggaaaagc tgggtgtcgc ctgcagagag atatgcagtt gttcctgatg 1861 aaacgggcct aacggatggt tcaagcaaag gtaatggagg tgatatcagt gttccacaaa 1921 ctgatgtcaa gcgtgtacga attaagcttc gaattctgca gtcgacggta ccgcgggccc 1981 gggatccacc ggtcgccacc atggtgagca agggcgagga ggataacatg gccatcatca 2041 aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac gagttcgaga 2101 tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag ctgaaggtga 2161 ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc atgtacggct 2221 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg tccttccccg 2281 agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc 2341 aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc ggcaccaact 2401 tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc tcctccgagc 2461 ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg aagctgaagg 2521 acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc 2581 tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac gaggactaca 2641 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc 2701 tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt 2761 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc 2821 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 2881 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 2941 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa 3001 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata 3061 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac 3121 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc 3181 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa 3241 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg 3301 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg 3361 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag 3421 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 3481 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 3541 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa 3601 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 3661 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 3721 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 3781 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 3841 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt 3901 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 3961 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 4021 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 4081 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc 4141 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 4201 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 4261 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 4321 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 4381 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 4441 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga 4501 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 4561 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 4621 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 4681 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 4741 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 4801 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 4861 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct 4921 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag 4981 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg 5041 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc 5101 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg 5161 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact 5221 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 5281 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 5341 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 5401 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 5461 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca 5521 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 5581 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 5641 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 5701 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 5761 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 5821 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 5881 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 5941 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 6001 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat //