LOCUS       sequence-311116-        5561 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of PULSE-controlled LFY-VP16-2A-RLuc.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5561)
  AUTHORS   Ochoa-Fernandez R, Abel NB, Wieland FG, Schlegel J, Koch LA, Miller 
            JB, Engesser R, Giuriani G, Brandl SM, Timmer J, Weber W, Ott T, 
            Simon R, Zurbriggen MD
  TITLE     Optogenetic control of gene expression in plants in the presence of 
            ambient white light.
  JOURNAL   Nat Methods. 2020 Jul;17(7):717-725. doi: 10.1038/s41592-020-0868-y.
            Epub 2020 Jun 29.
  PUBMED    32601426
REFERENCE   2  (bases 1 to 5561)
  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"; doi: 
            "10.1038/s41592-020-0868-y"; journalName: "Nat Methods"; date: 
            "2020-07"; volume: "17"; issue: "7"; pages: "717-725"
COMMENT     Sequence Label: pROF394
FEATURES             Location/Qualifiers
     source          1..5561
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     458..482
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      573..582
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2034..2267
                     /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"
     CDS             2268..2288
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2406..3341
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    3406..3527
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3443..3462)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3497..3516
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3561..3578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3732..4320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3812..3831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4491..5282,5283..5351))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5114..5133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5352..5456)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gattgaatat aaccgacgtg actgttacat ttagggattg aatataaccg acgtgactgt
       61 tacatttagg gattgaatat aaccgacgtg actgttacat ttagggattg aatataaccg
      121 acgtgactgt tacatttagg gataaacgat tgaatataac cgacgtgact gttacattta
      181 gggattgaat ataaccgacg tgactgttac atttagggat tgaatataac cgacgtgact
      241 gttacattta gggattgaat ataaccgacg tgactgttac atttagggat gctagcctcg
      301 agtaggtagc ctttagtcca tgcgttatag gtagccttta gtccatgcgt tataggtagc
      361 ctttagtcca tgcgttatag gtagccttta gtccatgcgt tataggtagc ctttagtcca
      421 tgaagcttag acactagagc tagccctata taagcagagc tcgtttagtg aaccgtcaga
      481 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
      541 gcctccgcgg ccccggtacc gaattcgcgg ccgccaccat ggatcctgaa ggtttcacga
      601 gtggcttatt ccggtggaac ccaacgagag cattggttca agcaccacct ccggttccac
      661 ctccgctgca gcaacagccg gtgacaccgc agacggctgc ttttgggatg cgacttggtg
      721 gtttagaggg actattcggt ccgtacggta tacgtttcta cacggcggcg aagatagcgg
      781 agttaggttt tacggcgagc acgcttgtgg gtatgaagga cgaggagctt gaagagatga
      841 tgaatagtct ctctcatatc tttcgttggg agcttcttgt tggtgaacgg tacggtatca
      901 aagctgccgt tagagctgaa cggagacgat tgcaagaaga ggaggaagag gaatcttcta
      961 gacgccgtca tttgctactc tccgccgctg gtgattccgg tactcatcac gctcttgatg
     1021 ctctctccca agaagggtta tctgaggaac cggtgcagca acaagaccag actgatgcgg
     1081 cggggaataa cggcggagga ggaagtggtt actgggacgc aggtcaagga aagatgaaga
     1141 agcaacagca gcagagacgg agaaagaaac caatgctgac gtcagtggaa accgacgaag
     1201 acgtcaacga aggtgaggat gacgacggga tggataacgg caacggaggt agtggtttgg
     1261 ggacagagag acagagggag catccgttta tcgtaacgga gcctggggaa gtggcacgtg
     1321 gcaaaaagaa cggcttagat tatctgttcc acttgtacga acaatgccgt gagttccttc
     1381 ttcaggtcca gacaattgct aaagaccgtg gcgaaaaatg ccccaccaag gtgacgaacc
     1441 aagtattcag gtacgcgaag aaatcaggag cgagttacat aaacaagcct aaaatgcgac
     1501 actacgttca ctgttacgct ctccactgcc tagacgaaga agcttcaaat gctctcagaa
     1561 gagcgtttaa agaacgcggt gagaacgttg gctcatggcg tcaggcttgt tacaagccac
     1621 ttgtgaacat cgcttgtcgt catggctggg atatagacgc cgtctttaac gctcatcctc
     1681 gtctctctat ttggtatgtt ccaacaaagc tgcgtcagct ttgccatttg gagcggaaca
     1741 atgcggttgc tgcggctgcg gctttagttg gcggtattag ctgtaccgga tcgtcgacgt
     1801 ctggacgtgg tggatgcggc ggcgacgact tgcgtttcac tagtgaattc gatagtgctg
     1861 gtagtgctgg tagtgctggt tccgcgtaca gccgcgcgcg tacgaaaaac aattacgggt
     1921 ctaccatcga gggcctgctc gatctcccgg acgacgacgc ccccgaagag gcggggctgg
     1981 cggctccgcg cctgtccttt ctccccgcgg gacacacgcg cagactgtcg acggcccccc
     2041 cgaccgatgt cagcctgggg gacgagctcc acttagacgg cgaggacgtg gcgatggcgc
     2101 atgccgacgc gctagacgat ttcgatctgg acatgttggg ggacggggat tccccgggtc
     2161 cgggatttac cccccacgac tccgccccct acggcgctct ggatatggcc gacttcgagt
     2221 ttgagcagat gtttaccgat gcccttggaa ttgacgagta cggtgggccc aagaaaaagc
     2281 ggaaggtgcc cgtgaaacag ctgctcaact tcgatctcct caaactggcc ggcgacgtgg
     2341 aatcaaatcc tggacccgcg cgcggtgcag gcgctggagc cggtgccggg gcaggcgctg
     2401 gcgctatgac ttcgaaagtt tatgatccag aacaaaggaa acggatgata actggtccgc
     2461 agtggtgggc cagatgtaaa caaatgaatg ttcttgattc atttattaat tattatgatt
     2521 cagaaaaaca tgcagaaaat gctgttattt ttttacatgg taacgcggcc tcttcttatt
     2581 tatggcgaca tgttgtgcca catattgagc cagtagcgcg gtgtattata ccagacctta
     2641 ttggtatggg caaatcaggc aaatctggta atggttctta taggttactt gatcattaca
     2701 aatatcttac tgcatggttt gaacttctta atttaccaaa gaagatcatt tttgtcggcc
     2761 atgattgggg tgcttgtttg gcatttcatt atagctatga gcatcaagat aagatcaaag
     2821 caatagttca cgctgaaagt gtagtagatg tgattgaatc atgggatgaa tggcctgata
     2881 ttgaagaaga tattgcgttg atcaaatctg aagaaggaga aaaaatggtt ttggagaata
     2941 acttcttcgt ggaaaccatg ttgccatcaa aaatcatgag aaagttagaa ccagaagaat
     3001 ttgcagcata tcttgaacca ttcaaagaga aaggtgaagt tcgtcgtcca acattatcat
     3061 ggcctcgtga aatcccgtta gtaaaaggtg gtaaacctga cgttgtacaa attgttagga
     3121 attataatgc ttatctacgt gcaagtgatg atttaccaaa aatgtttatt gaatcggacc
     3181 caggattctt ttccaatgct attgttgaag gtgccaagaa gtttcctaat actgaatttg
     3241 tcaaagtaaa aggtcttcat ttttcgcaag aagatgcacc tgatgaaatg ggaaaatata
     3301 tcaaatcgtt cgttgagcga gttctcaaaa atgaacaata aactagtgcg atcgcttaat
     3361 taatttaaat acgcgtttaa acagatccag atcttattaa agcagaactt gtttattgca
     3421 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3481 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggtcg
     3541 actctagact cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3601 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3661 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3721 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3781 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3841 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3901 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3961 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4021 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4081 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4141 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4201 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4261 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4321 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4381 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4441 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4501 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4561 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4621 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4681 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4741 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4801 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4861 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4921 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4981 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5041 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5101 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5161 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5221 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5281 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5341 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5401 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5461 tttccccgaa aagtgccacc tgacgtcgtc gacgatcgac ctgcaggtag gcgtgtacgg
     5521 tgggaggcct aggcttttgc aaaaagcttc gaagtttaaa c
//
