LOCUS       sequence-311119-        6910 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Constitutive expression LFY-VP16-2A-RLuc in plants.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6910)
  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 6910)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pROF417
FEATURES             Location/Qualifiers
     source          1..6910
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..346
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     289..309
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     regulatory      375..384
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1836..2069
                     /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             2070..2090
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2208..3143
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="MTSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     terminator      3212..3464
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(3515..3533)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3554..3570)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3554..3570)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3567..3589)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3578..3594
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3602..3608,3609..3626,3627..3632))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3647..3668
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3785..3802)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3956..4544)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4036..4055)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4715..5506,5507..5575))
                     /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     5338..5357
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5576..5680)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5706..6161)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5843..5864)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6055..6074
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6288..6310
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6302..6319
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6303..6319
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6326..6345
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6326..6344
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tgagactttt caacaaaggg taatatccgg aaacctcctc ggattccatt gcccagctat
       61 ctgtcacttt attgtgaaga tagtggaaaa ggaaggtggc tcctacaaat gccatcattg
      121 cgataaagga aaggccatcg ttgaagatgc ctctgccgac agtggtccca aagatggacc
      181 cccacccacg aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt
      241 ggattgatgt gatatcacca ctgacgtaag ggatgacgca caatcccact atccttcgca
      301 agacccttcc tctatataag gaagttcatt tcatttggag agaacacggg ggactctagc
      361 gctccggtgc ggccgccacc atggatcctg aaggtttcac gagtggctta ttccggtgga
      421 acccaacgag agcattggtt caagcaccac ctccggttcc acctccgctg cagcaacagc
      481 cggtgacacc gcagacggct gcttttggga tgcgacttgg tggtttagag ggactattcg
      541 gtccgtacgg tatacgtttc tacacggcgg cgaagatagc ggagttaggt tttacggcga
      601 gcacgcttgt gggtatgaag gacgaggagc ttgaagagat gatgaatagt ctctctcata
      661 tctttcgttg ggagcttctt gttggtgaac ggtacggtat caaagctgcc gttagagctg
      721 aacggagacg attgcaagaa gaggaggaag aggaatcttc tagacgccgt catttgctac
      781 tctccgccgc tggtgattcc ggtactcatc acgctcttga tgctctctcc caagaagggt
      841 tatctgagga accggtgcag caacaagacc agactgatgc ggcggggaat aacggcggag
      901 gaggaagtgg ttactgggac gcaggtcaag gaaagatgaa gaagcaacag cagcagagac
      961 ggagaaagaa accaatgctg acgtcagtgg aaaccgacga agacgtcaac gaaggtgagg
     1021 atgacgacgg gatggataac ggcaacggag gtagtggttt ggggacagag agacagaggg
     1081 agcatccgtt tatcgtaacg gagcctgggg aagtggcacg tggcaaaaag aacggcttag
     1141 attatctgtt ccacttgtac gaacaatgcc gtgagttcct tcttcaggtc cagacaattg
     1201 ctaaagaccg tggcgaaaaa tgccccacca aggtgacgaa ccaagtattc aggtacgcga
     1261 agaaatcagg agcgagttac ataaacaagc ctaaaatgcg acactacgtt cactgttacg
     1321 ctctccactg cctagacgaa gaagcttcaa atgctctcag aagagcgttt aaagaacgcg
     1381 gtgagaacgt tggctcatgg cgtcaggctt gttacaagcc acttgtgaac atcgcttgtc
     1441 gtcatggctg ggatatagac gccgtcttta acgctcatcc tcgtctctct atttggtatg
     1501 ttccaacaaa gctgcgtcag ctttgccatt tggagcggaa caatgcggtt gctgcggctg
     1561 cggctttagt tggcggtatt agctgtaccg gatcgtcgac gtctggacgt ggtggatgcg
     1621 gcggcgacga cttgcgtttc actagtgaat tcgatagtgc tggtagtgct ggtagtgctg
     1681 gttccgcgta cagccgcgcg cgtacgaaaa acaattacgg gtctaccatc gagggcctgc
     1741 tcgatctccc ggacgacgac gcccccgaag aggcggggct ggcggctccg cgcctgtcct
     1801 ttctccccgc gggacacacg cgcagactgt cgacggcccc cccgaccgat gtcagcctgg
     1861 gggacgagct ccacttagac ggcgaggacg tggcgatggc gcatgccgac gcgctagacg
     1921 atttcgatct ggacatgttg ggggacgggg attccccggg tccgggattt accccccacg
     1981 actccgcccc ctacggcgct ctggatatgg ccgacttcga gtttgagcag atgtttaccg
     2041 atgcccttgg aattgacgag tacggtgggc ccaagaaaaa gcggaaggtg cccgtgaaac
     2101 agctgctcaa cttcgatctc ctcaaactgg ccggcgacgt ggaatcaaat cctggacccg
     2161 cgcgcggtgc aggcgctgga gccggtgccg gggcaggcgc tggcgctatg acttcgaaag
     2221 tttatgatcc agaacaaagg aaacggatga taactggtcc gcagtggtgg gccagatgta
     2281 aacaaatgaa tgttcttgat tcatttatta attattatga ttcagaaaaa catgcagaaa
     2341 atgctgttat ttttttacat ggtaacgcgg cctcttctta tttatggcga catgttgtgc
     2401 cacatattga gccagtagcg cggtgtatta taccagacct tattggtatg ggcaaatcag
     2461 gcaaatctgg taatggttct tataggttac ttgatcatta caaatatctt actgcatggt
     2521 ttgaacttct taatttacca aagaagatca tttttgtcgg ccatgattgg ggtgcttgtt
     2581 tggcatttca ttatagctat gagcatcaag ataagatcaa agcaatagtt cacgctgaaa
     2641 gtgtagtaga tgtgattgaa tcatgggatg aatggcctga tattgaagaa gatattgcgt
     2701 tgatcaaatc tgaagaagga gaaaaaatgg ttttggagaa taacttcttc gtggaaacca
     2761 tgttgccatc aaaaatcatg agaaagttag aaccagaaga atttgcagca tatcttgaac
     2821 cattcaaaga gaaaggtgaa gttcgtcgtc caacattatc atggcctcgt gaaatcccgt
     2881 tagtaaaagg tggtaaacct gacgttgtac aaattgttag gaattataat gcttatctac
     2941 gtgcaagtga tgatttacca aaaatgttta ttgaatcgga cccaggattc ttttccaatg
     3001 ctattgttga aggtgccaag aagtttccta atactgaatt tgtcaaagta aaaggtcttc
     3061 atttttcgca agaagatgca cctgatgaaa tgggaaaata tatcaaatcg ttcgttgagc
     3121 gagttctcaa aaatgaacaa taagaattcc ccgggctcga gaagcttgga tccaccggat
     3181 ctagataact gatctcgagg cgaatttccc cgatcgttca aacatttggc aataaagttt
     3241 cttaagattg aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatta
     3301 cgttaagcat gtaataatta acatgtaatg catgacgtta tttatgagat gggtttttat
     3361 gattagagtc ccgcaattat acatttaata cgcgatagaa aacaaaatat agcgcgcaaa
     3421 ctaggataaa ttatcgcgcg cggtgtcatc tatgttacta gatcgggaat taattcgata
     3481 agctagagat ccgtcaaggt accagctttt gttcccttta gtgagggtta atttcgagct
     3541 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3601 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3661 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3721 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3781 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3841 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3901 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3961 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4021 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4081 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4141 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4201 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4261 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4321 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4381 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4441 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4501 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4561 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4621 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4681 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4741 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4801 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4861 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4921 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4981 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5041 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5101 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5161 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5221 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5281 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5341 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5401 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5461 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5521 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5581 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5641 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5701 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     5761 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5821 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5881 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5941 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     6001 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     6061 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     6121 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     6181 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     6241 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     6301 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcca
     6361 ccgcggagcg gagaattaag ggagtcaggc cttaattaag agctcgcatg cctgcaggtc
     6421 cccagattag ccttttcaat ttcagaaaga atgctaaccc acagatggtt agagaggctt
     6481 acgcagcagg tctcatcaag acgatctacc cgagcaataa tctccaggaa atcaaatacc
     6541 ttcccaagaa ggttaaagat gcagtcaaaa gattcaggac taactgcatc aagaacacag
     6601 agaaagatat atttctcaag atcagaagta ctattccagt atggacgatt caaggcttgc
     6661 ttcacaaacc aaggcaagta atagagattg gagtctctaa aaaggtagtt cccactgaat
     6721 caaaggccat ggagtcaaag attcaaatag aggacctaac agaactcgcc gtaaagactg
     6781 gcgaacagtt catacagagt ctcttacgac tcaatgacaa gaagaaaatc ttcgtcaaca
     6841 tggtggagca cgacacactt gtctactcca aaaatatcaa agatacagtc tcagaagacc
     6901 aaagggcaat
//
