LOCUS       sequence-408987-        6159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of LOV V416L, a mutant with super long 
            relaxation time.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6159)
  AUTHORS   Wang H, Vilela M, Winkler A, Tarnawski M, Schlichting I, Yumerefendi
            H, Kuhlman B, Liu R, Danuser G, Hahn KM
  TITLE     LOVTRAP: an optogenetic system for photoinduced protein 
            dissociation.
  JOURNAL   Nat Methods. 2016 Jul 18. doi: 10.1038/nmeth.3926.
  PUBMED    27427858
REFERENCE   2  (bases 1 to 6159)
  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. 2016 Jul 18. doi: 10.1038/nmeth.3926."
COMMENT     Sequence Label: pTriEx-mCherry-LOV2 V416L
FEATURES             Location/Qualifiers
     source          1..6159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     249..271
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     333..352
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     394..413
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        394..412
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    413..437
                     /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        453..562
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             577..1284
                     /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(722..740)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             1297..1725
                     /codon_start=1
                     /gene="NPH1 from Avena sativa"
                     /product="light, oxygen, or voltage domain from phototropin
                     1 of Avena sativa (Herrou and Crosson, 2011)"
                     /label=LOV2
                     /note="photoswitch that undergoes a light-inducible 
                     conformational change"
                     /translation="LATTLERIEKNFLITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKEL"
     CDS             1765..1797
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             1804..1827
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(1917..1936)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1982..2037
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2036..2055)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      2125..2172
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     2185..2890
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           2273..2645
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(2302..2890)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(2899..2932)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(3100..3687)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3179..3198)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3700..4491,4492..4560))
                     /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     4323..4342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_recomb     4803..5630
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4803..4927
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4965..5570)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        5698..6001
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6003..6159,1..46)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(6156..6159,1..17)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agacgtcgtt tagtgaaccg
       61 tcagatcact agatgcttta ttgcggtagt ttatcacagt taaattgcta acgccagtct
      121 cgaacttaac gtgcagaagt tggtcgtgag gcactgggca ggtaagtatc gggccctttg
      181 tgcgggggga gcggctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg
      241 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt
      301 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca
      361 tcattttggc aaagaattgg atcggaccga aattaatacg actcactata ggggaattgt
      421 gagcggataa caattccccg gagttaatcc gggaccttta attcaaccca acacaatata
      481 ttatagttaa ataagaatta ttatcaaatc atttgtatat taattaaaat actatactgt
      541 aaattacatt ttatttacaa tcaaaggaga tataccatgg tgagcaaggg cgaggaggat
      601 aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac
      661 ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc
      721 gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct
      781 cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg
      841 aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc
      901 gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag
      961 ctgcgcggca ccaacttccc ctccgacggc cctgtaatgc agaagaagac tatgggctgg
     1021 gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag
     1081 aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc
     1141 aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc
     1201 cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc
     1261 ggcggcatgg acgagctgta caagggttct ggatccttgg ctactacact tgaacgtatt
     1321 gagaagaact ttctcattac tgacccaaga ttgccagata atcccattat attcgcgtcc
     1381 gatagtttct tgcagttgac agaatatagc cgtgaagaaa ttttgggaag aaactgcagg
     1441 tttctacaag gtcctgaaac tgatcgcgcg acagtgagaa aaattagaga tgccatagat
     1501 aaccaaacag aggtcactgt tcagctgatt aattatacaa agagtggtaa aaagttctgg
     1561 aacctctttc acttgcagcc tatgcgagat cagaagggag atgtccagta ctttattggg
     1621 gttcagttgg atggaactga gcatgtccga gatgctgccg agagagaggg agtcatgctg
     1681 attaagaaaa ctgcagaaaa tattgatgag gcggcaaaag aactttaaaa gcttgcggcc
     1741 gcacagctgt atacacgtgc aagccagcca gaactcgccc cggaagaccc cgaggatctc
     1801 gagcaccacc atcaccatca ccatcactaa gtgattaacc tcaggtgcag gctgcctatc
     1861 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatcgatctt
     1921 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     1981 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     2041 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     2101 ttggcaacat atgcccatat gtaactagca taaccccttg gggcctctaa acgggtcttg
     2161 aggggttttt tgctgaaagc atgcggagga aattctcctt gaagtttccc tggtgttcaa
     2221 agtaaaggag tttgcaccag acgcacctct gttcactggt ccggcgtatt aaaacacgat
     2281 acattgttat tagtacattt attaagcgct agattctgtg cgttgttgat ttacagacaa
     2341 ttgttgtacg tattttaata attcattaaa tttataatct ttagggtggt atgttagagc
     2401 gaaaatcaaa tgattttcag cgtctttata tctgaattta aatattaaat cctcaataga
     2461 tttgtaaaat aggtttcgat tagtttcaaa caagggttgt ttttccgaac cgatggctgg
     2521 actatctaat ggattttcgc tcaacgccac aaaacttgcc aaatcttgta gcagcaatct
     2581 agctttgtcg atattcgttt gtgttttgtt ttgtaataaa ggttcgacgt cgttcaaaat
     2641 attatgcgct tttgtatttc tttcatcact gtcgttagtg tacaattgac tcgacgtaaa
     2701 cacgttaaat agagcttgga catatttaac atcgggcgtg ttagctttat taggccgatt
     2761 atcgtcgtcg tcccaaccct cgtcgttaga agttgcttcc gaagacgatt ttgccatagc
     2821 cacacgacgc ctattaattg tgtcggctaa cacgtccgcg atcaaatttg tagttgagct
     2881 ttttggaatt gcgatcgcat aacttcgtat agcatacatt atacgaagtt ataagctcgg
     2941 aacgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3001 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3061 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgccccct
     3121 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3181 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3241 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3301 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3361 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3421 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3481 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3541 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3601 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3661 attacgcgca gaaaaaaagg atctcaagaa gatcctttgt taccaatgct taatcagtga
     3721 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3781 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3841 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3901 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3961 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     4021 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     4081 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     4141 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     4201 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4261 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     4321 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4381 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4441 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4501 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4561 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgtccgcgcg
     4621 tttcctgcat cttttaatca aatcccaaga tgtgtataaa cgcgccggta tgtacaggaa
     4681 gaggtttata ctaaactgtt acattgcaaa cgtggtttcg tgtgccaagt gtgaaaaccg
     4741 atgtttaatc aaggctctga cgcatttcta caaccacgac tccaagtgtg tgggtgaagt
     4801 catgcatctt ttaatcaaat cccaagatgt gtataaacca ccaaactgcc aaaaaatgaa
     4861 aactgtcgac aagctctgtc cgtttgctgg caactgcaag ggtctcaatc ctatttgtaa
     4921 ttattgaata ataaaacaat tataaatgtc aaatttgttt tttattaacg atacaaacca
     4981 aacgcaacaa gaacatttgt agtattatct ataattgaaa acgcgtagtt ataatcgctg
     5041 aggtaatatt taaaatcatt ttcaaatgat tcacagttaa tttgcgacaa tataatttta
     5101 ttttcacata aactagacgc cttgtcgtct tcttcttcgt attccttctc tttttcattt
     5161 ttctcttcat aaaaattaac atagttatta tcgtatccat atatgtatct atcgtataga
     5221 gtaaattttt tgttgtcata aatatatatg tcttttttaa tggggtgtat agtaccgctg
     5281 cgcatagttt ttctgtaatt tacaacagtg ctattttctg gtagttcttc ggagtgtgtt
     5341 gctttaatta ttaaatttat ataatcaatg aatttgggat cgtcggtttt gtacaatatg
     5401 ttgccggcat agtacgcagc ttcttctagt tcaattacac cattttttag cagcaccgga
     5461 ttaacataac tttccaaaat gttgtacgaa ccgttaaaca aaaacagttc acctcccttt
     5521 tctatactat tgtctgcgag cagttgtttg ttgttaaaaa taacagccat tgtaatgaga
     5581 cgcacaaact aatatcacaa actggaaatg tctatcaata tatagttgct gatggccggc
     5641 ctattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
     5701 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
     5761 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
     5821 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
     5881 tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     5941 gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     6001 gctgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact cacggggatt
     6061 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     6121 ctttccaaaa tgtcgtaata accccgcccc gttgacgca
//
