LOCUS       sequence-177060-        3486 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gal4VP16 driven expression of NanoLuc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3486)
  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 3486)
  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-NanoLuc
FEATURES             Location/Qualifiers
     source          1..3486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    80..174
                     /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        193..431
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     primer_bind     378..399
                     /label=pCasper-hs
                     /note="Drosophila Hsp70 promoter, forward primer"
     regulatory      470..479
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             476..991
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="MVFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQR
                     IVLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGV
                     TPNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGW
                     RLCERILA"
     polyA_signal    1035..1156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1046..1065)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1100..1119
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1404..1421)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1575..2163)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1655..1674)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2363..3154,3155..3223))
                     /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    3328..3376
                     /note="synthetic polyadenylation signal"
     misc_feature    3390..3481
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3430..3449
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 aactggccgg tacctgagct cgctagcctc gaggatatca agatctggcc tcggcggcca
       61 agcttgcatg cctgcaggtc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg
      121 agtactgtcc tccgagcgga gtactgtcct ccgagcggag tactgtcctc cgagcggaga
      181 ctctagcgag cgccggagta taaatagagg cgcttcgtct acggagcgac aattcaattc
      241 aaacaagcaa agtgaacacg tcgctaagcg aaagctaagc aaataaacaa gcgcagctga
      301 acaagctaaa caatctgcag taaagtgcaa gttaaagtga atcaattaaa agtaaccagc
      361 aaccaagtaa atcaactgca actactgaaa tctgccaaga agtaattatt gaatacaaga
      421 agagaactct gaatagggaa ttgggaattc gttaacagat ccgatatccg ccaccatggt
      481 cttcacactc gaagatttcg ttggggactg gcgacagaca gccggctaca acctggacca
      541 agtccttgaa cagggaggtg tgtccagttt gtttcagaat ctcggggtgt ccgtaactcc
      601 gatccaaagg attgtcctga gcggtgaaaa tgggctgaag atcgacatcc atgtcatcat
      661 cccgtatgaa ggtctgagcg gcgaccaaat gggccagatc gaaaaaattt ttaaggtggt
      721 gtaccctgtg gatgatcatc actttaaggt gatcctgcac tatggcacac tggtaatcga
      781 cggggttacg ccgaacatga tcgactattt cggacggccg tatgaaggca tcgccgtgtt
      841 cgacggcaaa aagatcactg taacagggac cctgtggaac ggcaacaaaa ttatcgacga
      901 gcgcctgatc aaccccgacg gctccctgct gttccgagta accatcaacg gagtgaccgg
      961 ctggcggctg tgcgaacgca ttctggcgta ataattctag agtcggggcg gccggccgct
     1021 tcgagcagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     1081 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     1141 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
     1201 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga
     1261 tccgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     1321 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     1381 tgccggcagc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1441 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1501 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     1561 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     1621 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1681 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     1741 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     1801 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     1861 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     1921 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     1981 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2041 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2101 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2161 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2221 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     2281 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagcggccgc
     2341 aaatgctaaa ccactgcagt ggttaccagt gcttgatcag tgaggcaccg atctcagcga
     2401 tctgcctatt tcgttcgtcc atagtggcct gactccccgt cgtgtagatc actacgattc
     2461 gtgagggctt accatcaggc cccagcgcag caatgatgcc gcgagagccg cgttcaccgg
     2521 cccccgattt gtcagcaatg aaccagccag cagggagggc cgagcgaaga agtggtcctg
     2581 ctactttgtc cgcctccatc cagtctatga gctgctgtcg tgatgctaga gtaagaagtt
     2641 cgccagtgag tagtttccga agagttgtgg ccattgctac tggcatcgtg gtatcacgct
     2701 cgtcgttcgg tatggcttcg ttcaactctg gttcccagcg gtcaagccgg gtcacatgat
     2761 cacccatatt atgaagaaat gcagtcagct ccttagggcc tccgatcgtt gtcagaagta
     2821 agttggccgc ggtgttgtcg ctcatggtaa tggcagcact acacaattct cttaccgtca
     2881 tgccatccgt aagatgcttt tccgtgaccg gcgagtactc aaccaagtcg ttttgtgagt
     2941 agtgtatacg gcgaccaagc tgctcttgcc cggcgtctat acgggacaac accgcgccac
     3001 atagcagtac tttgaaagtg ctcatcatcg ggaatcgttc ttcggggcgg aaagactcaa
     3061 ggatcttgcc gctattgaga tccagttcga tatagcccac tcttgcaccc agttgatctt
     3121 cagcatcttt tactttcacc agcgtttcgg ggtgtgcaaa aacaggcaag caaaatgccg
     3181 caaagaaggg aatgagtgcg acacgaaaat gttggatgct catactcgtc ctttttcaat
     3241 attattgaag catttatcag ggttactagt acgtctctca aggataagta agtaatatta
     3301 aggtacggga ggtattggac aggccgcaat aaaatatctt tattttcatt acatctgtgt
     3361 gttggttttt tgtgtgaatc gatagtacta acatacgctc tccatcaaaa caaaacgaaa
     3421 caaaacaaac tagcaaaata ggctgtcccc agtgcaagtg caggtgccag aacatttctc
     3481 tggcct
//
