LOCUS Exported 5986 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pKeratin-PSmOrange2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5986) AUTHORS Subach OM, Entenberg D, Condeelis JS, Verkhusha VV TITLE A FRET-facilitated photoswitching using an orange fluorescent protein with the fast photoconversion kinetics. JOURNAL J Am Chem Soc. 2012 Aug 17. PUBMED 22900938 REFERENCE 2 (bases 1 to 5986) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5986 /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 1935..1944 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1941..2648 /codon_start=1 /product="enhanced orange to far-red photoswitchable monomeric fluorescent protein (Subach et al., 2012)" /label=PSmOrange2 /note="mammalian codon-optimized" /translation="MVSKGEENNMAIIKEFMRFKVHMEGTVNGHEFEIEGEGEGHPYEG FQTAKLKVTKGGPLPFAWDILSPLITYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY EDGGVVTVTQDSSLQDGEFIYKVKMRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIRMRLKLKDGGHYTSEVKTTYKAKKSVLLPGAYIVGIKLDITSHNEDYTIVEQYERS EARHSTGGMDELYK" primer_bind 2548..2571 /label=DsRed1-C /note="DsRed1, forward primer" polyA_signal 2774..2895 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2811..2830) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2865..2884 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2902..3357) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3039..3060) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3251..3270 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3384..3488 /gene="bla" /label=AmpR promoter promoter 3490..3847 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3513..3533) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3698..3833 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3760..3779 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3882..4676 /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(3936..3955) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4546..4565 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4864..4883) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4908..4955 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5284..5872 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5773..5792 /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 gctagcacca 601 tgagcttcac cactcgctcc accttctcca ccaactaccg gtccctgggc tctgtccagg 661 cgcccagcta cggcgcccgg ccggtcagca gcgcggccag cgtctatgca ggcgctgggg 721 gctctggttc ccggatctcc gtgtcccgct ccaccagctt caggggcggc atggggtccg 781 ggggcctggc caccgggata gccgggggtc tggcaggaat gggaggcatc cagaacgaga 841 aggagaccat gcaaagcctg aacgaccgcc tggcctctta cctggacaga gtgaggagcc 901 tggagaccga gaaccggagg ctggagagca aaatccggga gcacttggag aagaagggac 961 cccaggtcag agactggagc cattacttca agatcatcga ggacctgagg gctcagatct 1021 tcgcaaatac tgtggacaat gcccgcatcg ttctgcagat tgacaatgcc cgtcttgctg 1081 ctgatgactt tagagtcaag tatgagacag agctggccat gcgccagtct gtggagaacg 1141 acatccatgg gctccgcaag gtcattgatg acaccaatat cacacgactg cagctggaga 1201 cagagatcga ggctctcaag gaggagctgc tcttcatgaa gaagaaccac gaagaggaag 1261 taaaaggcct acaagcccag attgccagct ctgggttgac cgtggaggta gatgccccca 1321 aatctcagga cctcgccaag atcatggcag acatccgggc ccaatatgac gagctggctc 1381 ggaagaaccg agaggagcta gacaagtact ggtctcagca gattgaggag agcaccacag 1441 tggtcaccac acagtctgct gaggttggag ctgctgagac gacgctcaca gagctgagac 1501 gtacagtcca gtccttggag atcgacctgg actccatgag aaatctgaag gccagcttgg 1561 agaacagcct gagggaggtg gaggcccgct acgccctaca gatggagcag ctcaacggga 1621 tcctgctgca ccttgagtca gagctggcac agacccgggc agagggacag cgccaggccc 1681 aggagtatga ggccctgctg aacatcaagg tcaagctgga ggctgagatc gccacctacc 1741 gccgcctgct ggaagatggc gaggacttta atcttggtga tgccttggac agcagcaact 1801 ccatgcaaac catccaaaag accaccaccc gccggatagt ggatggcaaa gtggtgtctg 1861 agaccaatga caccaaagtt ctgaggcatc gaattctgca gtcgacggta ccgcgggccc 1921 gggatccacc ggtcgccacc atggtgagca agggcgagga gaataatatg gccatcatta 1981 aggagttcat gcgcttcaag gtgcacatgg agggcactgt gaacggccac gagttcgaga 2041 tcgagggcga gggcgagggc cacccctacg agggctttca gaccgctaag ctgaaggtga 2101 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc ccctctcatc acctacggct 2161 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttcaagctg tccttccccg 2221 agggcttcaa gtgggagcgc gtgatgaact acgaggacgg cggcgtggtg accgtgaccc 2281 aggactcctc tctgcaggac ggcgagttca tctacaaggt gaagatgcgc ggcaccaact 2341 tcccctccga cggccccgtg atgcagaaga agaccatggg ctgggaggcc tcctccgagc 2401 ggatgtaccc cgaggacggc gccctgaagg gcgagatcag gatgaggctg aagctgaagg 2461 atggcggcca ctacacctcc gaggtcaaga ctacctacaa ggccaagaag tccgtgctgc 2521 tgcccggcgc ctacatcgtc ggcatcaagc tggacatcac ctcccacaac gaggactaca 2581 ccatcgtgga acagtacgaa cgctccgagg cccgccactc caccggcggc atggacgagc 2641 tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt 2701 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc 2761 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 2821 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 2881 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa 2941 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata 3001 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac 3061 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc 3121 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa 3181 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg 3241 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg 3301 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag 3361 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt 3421 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa 3481 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa 3541 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 3601 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 3661 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 3721 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 3781 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt 3841 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat 3901 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac 3961 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc 4021 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc 4081 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag 4141 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc 4201 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg 4261 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc 4321 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc 4381 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga 4441 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca 4501 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg 4561 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg 4621 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg 4681 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga 4741 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg 4801 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct 4861 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag 4921 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg 4981 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc 5041 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg 5101 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact 5161 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat 5221 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc 5281 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct 5341 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg 5401 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca 5461 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc 5521 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga 5581 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac 5641 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga 5701 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag 5761 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg 5821 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag 5881 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc 5941 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat //