LOCUS       sequence-173095-        6777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PI(DM): Photo-activatable/light sensitive.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6777)
  AUTHORS   Dagliyan O, Tarnawski M, Chu PH, Shirvanyants D, Schlichting I, 
            Dokholyan NV, Hahn KM
  TITLE     Engineering extrinsic disorder to control protein activity in living
            cells.
  JOURNAL   Science. 2016 Dec 16;354(6318):1441-1444.
  PUBMED    27980211
REFERENCE   2  (bases 1 to 6777)
  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: "Science."; 
            date: "2016-12-16"; volume: "354(6318)"; pages: "1441-1444"
COMMENT     Sequence Label: pTriex-mVenus-PI-RhoA(Q63L)
FEATURES             Location/Qualifiers
     source          1..6777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     241..263
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     325..344
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     386..405
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        386..404
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    405..429
                     /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        445..554
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     CDS             575..592
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(599..601,602..1312)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(641..662)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(902..921)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1249..1270
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1478..1906
                     /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="LATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEILGR
                     NCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNLFHLQPMRDQKGDVQ
                     YFIGVQLDGTEHVRDAAEREGVMLIKKTAENIDEAAKEL"
     CDS             2375..2407
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             2414..2437
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     primer_bind     complement(2527..2546)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2592..2647
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2646..2665)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      2735..2782
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_recomb     2795..3500
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     3'UTR           2883..3255
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     CDS             complement(2912..3500)
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="IPKSSTTNLIADVLADTINRRRVAMAKSSSEATSNDEGWDDDDNR
                     PNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVEPLLQNKTQTNIDKAR
                     LLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDLIFKFRYKDAENHLIF
                     ALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
     protein_bind    complement(3509..3542)
                     /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(3710..4297)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3789..3808)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4310..5101,5102..5170))
                     /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     4933..4952
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     misc_recomb     5413..6240
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             5413..5537
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(5575..6180)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     enhancer        6308..6611
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6613..6777,1..38)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     join(6766..6777,1..9)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 gtaggcgtgt acggtgggag gtctatataa gcagacgtcg tttagtgaac cgtcagatca
       61 ctagatgctt tattgcggta gtttatcaca gttaaattgc taacgccagt ctcgaactta
      121 acgtgcagaa gttggtcgtg aggcactggg caggtaagta tcgggccctt tgtgcggggg
      181 gagcggctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
      241 ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct
      301 tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg
      361 gcaaagaatt ggatcggacc gaaattaata cgactcacta taggggaatt gtgagcggat
      421 aacaattccc cggagttaat ccgggacctt taattcaacc caacacaata tattatagtt
      481 aaataagaat tattatcaaa tcatttgtat attaattaaa atactatact gtaaattaca
      541 ttttatttac aatcaaagga gatataccat ggcacaccat caccaccatc acggttctgt
      601 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
      661 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
      721 gctgaccctg aagttcatca gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
      781 gaccaccctc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
      841 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
      901 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
      961 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1021 ggagtacaac tacaacagcc acaacgtcta tatcaccgcc gacaagcaga agaacggcat
     1081 caaggcaaac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgccgacca
     1141 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1201 gagctaccag tccaaactga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1261 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agggttctgg
     1321 aagtggatcc atggctgcca tccggaagaa actggtgatt gttggtgatg gagcctgtgg
     1381 aaagacatgc ttgctcatag tcttcagcaa ggaccagttc ccagaggtgt atgtgcccac
     1441 agtgtttgag aactatgtgg cagatatcga ggtggatttg gctactacac ttgaacgtat
     1501 tgagaagaac tttgtcatta ctgacccaag attgccagat aatcccatta tattcgcgtc
     1561 cgatagtttc ttgcagttga cagaatatag ccgtgaagaa attttgggaa gaaactgcag
     1621 gtttctacaa ggtcctgaaa ctgatcgcgc gacagtgaga aaaattagag atgccataga
     1681 taaccaaaca gaggtcactg ttcagctgat taattataca aagagtggta aaaagttctg
     1741 gaacctcttt cacttgcagc ctatgcgaga tcagaaggga gatgtccagt actttattgg
     1801 ggttcagttg gatggaactg agcatgtccg agatgctgcc gagagagagg gagtcatgct
     1861 gattaagaaa actgcagaaa atattgatga ggcggcaaaa gaacttaagc aggtagagtt
     1921 ggctttgtgg gacacagctg ggctggaaga ttatgatcgc ctgaggcccc tctcctaccc
     1981 agataccgat gttatactga tgtgtttttc catcgacagc cctgatagtt tagaaaacat
     2041 cccagaaaag tggaccccag aagtcaagca tttctgtccc aacgtgccca tcatcctggt
     2101 tgggaataag aaggatcttc ggaatgatga gcacacaagg cgggagctag ccaagatgaa
     2161 gcaggagccg gtgaaacctg aagaaggcag agatatggca aacaggattg gcgcttttgg
     2221 gtacatggag tgttcagcaa agaccaaaga tggagtgaga gaggtttttg aaatggctac
     2281 gagagctgct ctgcaagcta gacgtgggaa gaaaaaatct gggtgccttg tcttgtaaaa
     2341 gcttgcggcc gcacagctgt atacacgtgc aagccagcca gaactcgccc cggaagaccc
     2401 cgaggatctc gagcaccacc atcaccatca ccatcactaa gtgattaacc tcaggtgcag
     2461 gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga
     2521 gatcgatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     2581 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     2641 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     2701 ggtttagagt ttggcaacat atgcccatat gtaactagca taaccccttg gggcctctaa
     2761 acgggtcttg aggggttttt tgctgaaagc atgcggagga aattctcctt gaagtttccc
     2821 tggtgttcaa agtaaaggag tttgcaccag acgcacctct gttcactggt ccggcgtatt
     2881 aaaacacgat acattgttat tagtacattt attaagcgct agattctgtg cgttgttgat
     2941 ttacagacaa ttgttgtacg tattttaata attcattaaa tttataatct ttagggtggt
     3001 atgttagagc gaaaatcaaa tgattttcag cgtctttata tctgaattta aatattaaat
     3061 cctcaataga tttgtaaaat aggtttcgat tagtttcaaa caagggttgt ttttccgaac
     3121 cgatggctgg actatctaat ggattttcgc tcaacgccac aaaacttgcc aaatcttgta
     3181 gcagcaatct agctttgtcg atattcgttt gtgttttgtt ttgtaataaa ggttcgacgt
     3241 cgttcaaaat attatgcgct tttgtatttc tttcatcact gtcgttagtg tacaattgac
     3301 tcgacgtaaa cacgttaaat agagcttgga catatttaac atcgggcgtg ttagctttat
     3361 taggccgatt atcgtcgtcg tcccaaccct cgtcgttaga agttgcttcc gaagacgatt
     3421 ttgccatagc cacacgacgc ctattaattg tgtcggctaa cacgtccgcg atcaaatttg
     3481 tagttgagct ttttggaatt gcgatcgcat aacttcgtat agcatacatt atacgaagtt
     3541 ataagctcgg aacgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3601 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3661 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3721 tccgccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     3781 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     3841 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     3901 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     3961 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4021 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4081 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4141 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4201 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4261 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttgt taccaatgct
     4321 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4381 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4441 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4501 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     4561 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4621 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4681 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4741 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4801 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4861 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4921 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4981 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5041 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5101 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5161 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5221 tgtccgcgcg tttcctgcat cttttaatca aatcccaaga tgtgtataaa cgcgccggta
     5281 tgtacaggaa gaggtttata ctaaactgtt acattgcaaa cgtggtttcg tgtgccaagt
     5341 gtgaaaaccg atgtttaatc aaggctctga cgcatttcta caaccacgac tccaagtgtg
     5401 tgggtgaagt catgcatctt ttaatcaaat cccaagatgt gtataaacca ccaaactgcc
     5461 aaaaaatgaa aactgtcgac aagctctgtc cgtttgctgg caactgcaag ggtctcaatc
     5521 ctatttgtaa ttattgaata ataaaacaat tataaatgtc aaatttgttt tttattaacg
     5581 atacaaacca aacgcaacaa gaacatttgt agtattatct ataattgaaa acgcgtagtt
     5641 ataatcgctg aggtaatatt taaaatcatt ttcaaatgat tcacagttaa tttgcgacaa
     5701 tataatttta ttttcacata aactagacgc cttgtcgtct tcttcttcgt attccttctc
     5761 tttttcattt ttctcttcat aaaaattaac atagttatta tcgtatccat atatgtatct
     5821 atcgtataga gtaaattttt tgttgtcata aatatatatg tcttttttaa tggggtgtat
     5881 agtaccgctg cgcatagttt ttctgtaatt tacaacagtg ctattttctg gtagttcttc
     5941 ggagtgtgtt gctttaatta ttaaatttat ataatcaatg aatttgggat cgtcggtttt
     6001 gtacaatatg ttgccggcat agtacgcagc ttcttctagt tcaattacac cattttttag
     6061 cagcaccgga ttaacataac tttccaaaat gttgtacgaa ccgttaaaca aaaacagttc
     6121 acctcccttt tctatactat tgtctgcgag cagttgtttg ttgttaaaaa taacagccat
     6181 tgtaatgaga cgcacaaact aatatcacaa actggaaatg tctatcaata tatagttgct
     6241 gatggccggc ctattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     6301 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     6361 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6421 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6481 atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6541 cccagtacat gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg
     6601 ctattaccat gctgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact
     6661 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     6721 atcaacggga ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcg
//
