LOCUS Exported 3698 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gal4VP16 driven expression of mCherry-NLS in mammalian cells. ACCESSION . VERSION . KEYWORDS pUAS-mCherry-NLS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3698) AUTHORS Zhang W, Lohman AW, Zhuravlova Y, Lu X, Wiens MD, Hoi H, Yaganoglu S, Mohr MA, Kitova EN, Klassen JS, Pantazis P, Thompson RJ, Campbell RE TITLE Optogenetic control with a photocleavable protein, PhoCl. JOURNAL Nat Methods. 2017 Mar 13. doi: 10.1038/nmeth.4222. PUBMED 28288123 REFERENCE 2 (bases 1 to 3698) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3698 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 76..170 /label=5X UAS /bound_moiety="GAL4" /note="five tandem copies of the ""ScaI site"" 17-mer CGGAGTACTGTCCTCCG, an upstream activating sequence (UAS) that efficiently binds yeast Gal4 (Webster et al., 1988; Pfeiffer et al., 2010)" promoter 189..427 /label=hsp70 promoter /note="Drosophila melanogaster hsp70Bb promoter" primer_bind 374..395 /label=pCasper-hs /note="Drosophila Hsp70 promoter, forward primer" regulatory 466..475 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 472..1179 /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(617..635) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 885..904 /label=mCherry-F /note="mCherry, forward primer" CDS 1180..1200 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" polyA_signal 1243..1364 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1254..1273) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1308..1327 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(1612..1629) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1783..2371) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1863..1882) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2571..3431) /codon_start=1 /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" polyA_signal 3536..3584 /note="synthetic polyadenylation signal" misc_feature 3598..3689 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" primer_bind 3638..3657 /label=RVprimer3 /note="pGL3 vector, forward primer" ORIGIN 1 ggccggtacc tgagctcgct agcctcgagg atatcaagat ctggcctcgg cggccaagct 61 tgcatgcctg caggtcggag tactgtcctc cgagcggagt actgtcctcc gagcggagta 121 ctgtcctccg agcggagtac tgtcctccga gcggagtact gtcctccgag cggagactct 181 agcgagcgcc ggagtataaa tagaggcgct tcgtctacgg agcgacaatt caattcaaac 241 aagcaaagtg aacacgtcgc taagcgaaag ctaagcaaat aaacaagcgc agctgaacaa 301 gctaaacaat ctgcagtaaa gtgcaagtta aagtgaatca attaaaagta accagcaacc 361 aagtaaatca actgcaacta ctgaaatctg ccaagaagta attattgaat acaagaagag 421 aactctgaat agggaattgg gaattcgtta acagatccga tatccgccac catggtgagc 481 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg 541 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac 601 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg 661 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac 721 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac 781 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc 841 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag 901 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag 961 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag 1021 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag 1081 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag 1141 ggccgccact ccaccggcgg catggacgag ctgtacaagc caaagaaaaa gagaaaagta 1201 taatctagag tcggggcggc cggccgcttc gagcagacat gataagatac attgatgagt 1261 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg 1321 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca 1381 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc 1441 tctacaaatg tggtaaaatc gataaggatc cgtcgaccga tgcccttgag agccttcaac 1501 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc 1561 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tcttccgctt cctcgctcac 1621 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 1681 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 1741 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 1801 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 1861 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 1921 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 1981 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 2041 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 2101 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 2161 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 2221 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 2281 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 2341 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 2401 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 2461 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 2521 tgagtaaact tggtctgaca gcggccgcaa atgctaaacc actgcagtgg ttaccagtgc 2581 ttgatcagtg aggcaccgat ctcagcgatc tgcctatttc gttcgtccat agtggcctga 2641 ctccccgtcg tgtagatcac tacgattcgt gagggcttac catcaggccc cagcgcagca 2701 atgatgccgc gagagccgcg ttcaccggcc cccgatttgt cagcaatgaa ccagccagca 2761 gggagggccg agcgaagaag tggtcctgct actttgtccg cctccatcca gtctatgagc 2821 tgctgtcgtg atgctagagt aagaagttcg ccagtgagta gtttccgaag agttgtggcc 2881 attgctactg gcatcgtggt atcacgctcg tcgttcggta tggcttcgtt caactctggt 2941 tcccagcggt caagccgggt cacatgatca cccatattat gaagaaatgc agtcagctcc 3001 ttagggcctc cgatcgttgt cagaagtaag ttggccgcgg tgttgtcgct catggtaatg 3061 gcagcactac acaattctct taccgtcatg ccatccgtaa gatgcttttc cgtgaccggc 3121 gagtactcaa ccaagtcgtt ttgtgagtag tgtatacggc gaccaagctg ctcttgcccg 3181 gcgtctatac gggacaacac cgcgccacat agcagtactt tgaaagtgct catcatcggg 3241 aatcgttctt cggggcggaa agactcaagg atcttgccgc tattgagatc cagttcgata 3301 tagcccactc ttgcacccag ttgatcttca gcatctttta ctttcaccag cgtttcgggg 3361 tgtgcaaaaa caggcaagca aaatgccgca aagaagggaa tgagtgcgac acgaaaatgt 3421 tggatgctca tactcgtcct ttttcaatat tattgaagca tttatcaggg ttactagtac 3481 gtctctcaag gataagtaag taatattaag gtacgggagg tattggacag gccgcaataa 3541 aatatcttta ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac 3601 atacgctctc catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag 3661 tgcaagtgca ggtgccagaa catttctctg gcctaact //