LOCUS       sequence-176852-        3698 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gal4VP16 driven expression of mCherry-NLS in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2017 Mar 13. doi: 10.1038/nmeth.4222."
COMMENT     Sequence Label: pUAS-mCherry-NLS
FEATURES             Location/Qualifiers
     source          1..3698
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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(join(2571..3362,3363..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
//
