LOCUS Exported 5999 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses NLS-PpsR2-VP16 under CMV promoter. ACCESSION . VERSION . KEYWORDS pCMV-104 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5999) AUTHORS Kaberniuk AA, Shemetov AA, Verkhusha VV TITLE A bacterial phytochrome-based optogenetic system controllable with near-infrared light. JOURNAL Nat Methods. 2016 May 9. doi: 10.1038/nmeth.3864. PUBMED 27159085 REFERENCE 2 (bases 1 to 5999) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5999 /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 2160..2169 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2319..2552 /codon_start=1 /gene="UL48" /product="transcriptional activation domain of herpes simplex virus protein VP16 (Triezenberg et al., 1988; Cousens et al., 1989)" /label=VP16 AD /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG" primer_bind complement(2634..2652) /label=T7 Term /note="T7 terminator, reverse primer" polyA_signal 2787..2908 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2824..2843) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2878..2897 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2915..3370) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3052..3073) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 3264..3283 /label=F1ori-R /note="F1 origin, reverse primer" promoter 3397..3501 /gene="bla" /label=AmpR promoter promoter 3503..3860 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3526..3546) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3711..3846 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3773..3792 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3895..4689 /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(3949..3968) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4559..4578 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4877..4896) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4921..4968 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 5297..5885 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5786..5805 /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 atctgcgatc gccatgtcgc ggaggcggca cagctacgag aacgatggtg 661 gacaacctca caaaaggagg aagacgtctc caccggtggc gtcaaagtcc gttcatgccg 721 acatcaccct tctgctcgat atggagggtg tgattcgcga agccaccctg tctccgacga 781 tggcggccga gagcgtggac ggttggctgg ggcgtcgctg gagcgacatc gccggcgccg 841 aaggcggcga caaggttcgc cgcatggtcg aagacgcccg ccgcagcggc atctcggctt 901 tccgccagat caatcagcct ttcccgagcg gcgtcgaaat cccgatcgaa ttcaccacga 961 tgctgctggg cgaccgcacc ggcatgatcg cggtcggcaa gaacatgcag gcggtcaccg 1021 agctgcattc ccggctgatc gctgcgcagc aggcgatgga gcgcgactat tggcggttgc 1081 gtgaattgga gactcgctac cgcctggtgt tcgacgctgc cgccgatgcg gtgatgatcg 1141 tctccgccgg cgacatgcgc atcgtcgaag ccaaccgggc ggcggtgaat gcgatcagcc 1201 gcgtcgagcg cggcaatgac gaccttgcgg ggcgtgattt cctcgccgaa gtggcggctg 1261 ccgatcgcga tgcggtgcgc gacatgctgg cccaggtgcg tcagcgcggc accgcactca 1321 gcgtcctcgt tcatctcggc cgttacgacc gcgcctggat gctgcgcggt tcgctgatgt 1381 cgtccgagcg tcgtcaggtt ttcctgctgc acttcacccc ggtgaccacg actcccgcga 1441 tcgacgacgt cgacgatgat gccgtgctgc gcgggctgat cgatcgcatt cccgacgggt 1501 tcgtcgcact ggattcggaa ggcgtcgttc gtcacgccaa ccaggcgttt ctcgatctgg 1561 tccagatcgg ctccaagcct gcggcggtcg gacgatcgct gggcgtctgg atgggtcgtc 1621 cgggcgccga tctgtccagc ttgctgacgc tgctgcggcg ctacaagacg gtgcggctgt 1681 tccaaacgac gatccgcggc gagctcggca ccgagactga agtcgaggtc tcggccgtcg 1741 acggcgagga cgaccaatac atcggcgttc tgatgcgcaa tgtcgcgcga cgcctcgacg 1801 ctgcggacga ccacgatgcc ttgcgtcagg cgctcggccc gatcagcaag cagctcgggc 1861 gatcctcgct gcgcaagctg gtgaagaacg ccgtgagcat tgtcgagcag cactacgtga 1921 aggaagcgct gttgcgatcc aagggcaatc gcacggcaac tgccgaactg ctcggattga 1981 gccggcagag cctttatgca aaactcaaca gctacggctt cgacgacaaa ggtgtcgttg 2041 cttctgctgc cgacggtgca gagggcgcct cagacgacgc agaggataag cttagtgctg 2101 gtggtagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggtccggacg 2161 ccaccatggc gtacagccgc gcgcgtacga aaaacaatta cgggtctacc atcgagggcc 2221 tgctcgatct cccggacgac gacgcccccg aagaggcggg gctggcggct ccgcgcctgt 2281 cctttctccc cgcgggacac acgcgcagac tgtcgacggc ccccccgacc gatgtcagcc 2341 tgggggacga gctccactta gacggcgagg acgtggcgat ggcgcatgcc gacgcgctag 2401 acgatttcga tctggacatg ttgggggacg gggattcccc gggtccggga tttacccccc 2461 acgactccgc cccctacggc gctctggata tggccgactt cgagtttgag cagatgttta 2521 ccgatgccct tggaattgac gagtacggtg ggtaatctag agtcgacctg caggcatgca 2581 agctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga 2641 gcaataacta gcataacccc ttggggcggc cgcgactcta gatcataatc agccatacca 2701 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac 2761 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat 2821 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 2881 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta 2941 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc 3001 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg 3061 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat 3121 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc 3181 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag 3241 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg 3301 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta 3361 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt 3421 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa 3481 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt 3541 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3601 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa 3661 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc 3721 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg 3781 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 3841 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt 3901 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat 3961 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag 4021 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac 4081 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac 4141 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc 4201 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg 4261 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag 4321 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat 4381 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag 4441 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc 4501 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg 4561 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg 4621 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag 4681 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat 4741 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc 4801 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc 4861 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga 4921 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg 4981 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg 5041 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg 5101 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga 5161 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc 5221 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa 5281 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa 5341 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc 5401 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt 5461 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc 5521 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac 5581 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca 5641 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg 5701 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag 5761 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt 5821 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat 5881 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc 5941 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat //