LOCUS       sequence-13376-e        8422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Gal4 Activation Domain fused to CIBN (aa1-170) for 
            blue-light-induced transcription activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8422)
  AUTHORS   Kennedy MJ, Hughes RM, Peteya LA, Schwartz JW, Ehlers MD, Tucker CL
  TITLE     Rapid blue-light-mediated induction of protein interactions in 
            living cells.
  JOURNAL   Nat Methods. 2010 Dec;7(12):973-5. doi: 10.1038/nmeth.1524
  PUBMED    21037589
REFERENCE   2  (bases 1 to 8422)
  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/nmeth"; 
            journalName: "Nat Methods"; date: "2010-12"; volume: "7"; issue: 
            "12"; pages: "973-5"
COMMENT     Sequence Label: pGal4AD-CIBN
FEATURES             Location/Qualifiers
     source          1..8422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        771..1476
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1522..1542
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1558..1899
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     primer_bind     1905..1924
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1905..1923
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1939..2448
                     /codon_start=1
                     /product="N-terminal portion of the CIB1 transcription 
                     factor from Arabidopsis thaliana"
                     /label=CIBN
                     /note="binds to CRY2 (cryptochrome 2) that has absorbed 
                     blue light (Kennedy et al., 2010)"
                     /translation="MNGAIGGDLLLNFPDMSVLERQRAHLKYLNPTFDSPLAGFFADSS
                     MITGGEMDSYLSTAGLNLPMMYGETTVEGDSRLSISPETTLGTGNFKKRKFDTETKDCN
                     EKKKKMTMNRDDLVEEGEEEKSKITEQNNGSTKSIKKMKHKAKKEENNFSNDSSKVTKE
                     LEKTDYI"
     terminator      2850..3037
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(2905..2929)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3154..4248)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(4249..4654)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(4696..4712)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4696..4712)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4709..4731)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4720..4736
                     /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(4744..4750,4751..4768,4769..4774))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4789..4810
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    4865..4898
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4976..5009
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5078..5095)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5249..5837)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5329..5348)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6008..6799,6800..6868))
                     /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     6631..6650
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6869..6973)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7041..7059
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7097..7119)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7219..7238
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      7255..8419
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tgcatgcctg caggtcgaga tccgggatcg aagaaatgat ggtaaatgaa ataggaaatc
       61 aaggagcatg aaggcaaaag acaaatataa gggtcgaacg aaaaataaag tgaaaagtgt
      121 tgatatgatg tatttggctt tgcggcgccg aaaaaacgag tttacgcaat tgcacaatca
      181 tgctgactct gtggcggacc cgcgctcttg ccggcccggc gataacgctg ggcgtgaggc
      241 tgtgcccggc ggagtttttt gcgcctgcat tttccaaggt ttaccctgcg ctaaggggcg
      301 agattggaga agcaataaga atgccggttg gggttgcgat gatgacgacc acgacaactg
      361 gtgtcattat ttaagttgcc gaaagaacct gagtgcattt gcaacatgag tatactagaa
      421 gaatgagcca agacttgcga gacgcgagtt tgccggtggt gcgaacaata gagcgaccat
      481 gaccttgaag gtgagacgcg cataaccgct agagtacttt gaagaggaaa cagcaatagg
      541 gttgctacca gtataaatag acaggtacat acaacactgg aaatggttgt ctgtttgagt
      601 acgctttcaa ttcatttggg tgtgcacttt attatgttac aatatggaag ggaactttac
      661 acttctccta tgcacatata ttaattaaag tccaatgcta gtagagaagg ggggtaacac
      721 ccctccgcgc tcttttccga tttttttcta aaccgtggaa tatttcggat atccttttgt
      781 tgtttccggg tgtacaatat ggacttcctc ttttctggca accaaaccca tacatcggga
      841 ttcctataat accttcgttg gtctccctaa catgtaggtg gcggagggga gatatacaat
      901 agaacagata ccagacaaga cataatgggc taaacaagac tacaccaatt acactgcctc
      961 attgatggtg gtacataacg aactaatact gtagccctag acttgatagc catcatcata
     1021 tcgaagtttc actacccttt ttccatttgc catctattga agtaataata ggcgcatgca
     1081 acttcttttc tttttttttc ttttctctct cccccgttgt tgtctcacca tatccgcaat
     1141 gacaaaaaaa atgatggaag acactaaagg aaaaaattaa cgacaaagac agcaccaaca
     1201 gatgtcgttg ttccagagct gatgaggggt atctcgaagc acacgaaact ttttccttcc
     1261 ttcattcacg cacactactc tctaatgagc aacggtatac ggccttcctt ccagttactt
     1321 gaatttgaaa taaaaaaaag tttgctgtct tgctatcaag tataaataga cctgcaatta
     1381 ttaatctttt gtttcctcgt cattgttctc gttccctttc ttccttgttt ctttttctgc
     1441 acaatatttc aagctatacc aagcatacaa tcaactccaa gctttgcaaa gatggataaa
     1501 gcggaattaa ttcccgagcc tccaaaaaag aagagaaagg tcgaattggg taccgccgcc
     1561 aattttaatc aaagtgggaa tattgctgat agctcattgt ccttcacttt cactaacagt
     1621 agcaacggtc cgaacctcat aacaactcaa acaaattctc aagcgctttc acaaccaatt
     1681 gcctcctcta acgttcatga taacttcatg aataatgaaa tcacggctag taaaattgat
     1741 gatggtaata attcaaaacc actgtcacct ggttggacgg accaaactgc gtataacgcg
     1801 tttggaatca ctacagggat gtttaatacc actacaatgg atgatgtata taactatcta
     1861 ttcgatgatg aagatacccc accaaaccca aaaaaagaga tctttaatac gactcactat
     1921 agggcgagcg ccgaattcat gaatggagct ataggaggtg accttttgct caattttcct
     1981 gacatgtcgg tcctagagcg ccaaagggct cacctcaagt acctcaatcc cacctttgat
     2041 tctcctctcg ccggcttctt tgccgattct tcaatgatta ccggcggcga gatggacagc
     2101 tatctttcga ctgccggttt gaatcttccg atgatgtacg gtgagacgac ggtggaaggt
     2161 gattcaagac tctcaatttc gccggaaacg acgcttggga ctggaaattt caagaaacgg
     2221 aagtttgata cagagactaa ggattgtaat gagaagaaga agaagatgac gatgaacaga
     2281 gatgacctag tagaagaagg agaagaagag aagtcgaaaa taacagagca aaacaatggg
     2341 agcacaaaaa gcatcaagaa gatgaaacac aaagccaaga aagaagagaa caatttctct
     2401 aatgattcat ctaaagtgac gaaggaattg gagaaaacgg attatattca tggatccatc
     2461 gagctcgagc tgcagatgaa tcgtagatac tgaaaaaccc cgcaagttca cttcaactgt
     2521 gcatcgtgca ccatctcaat ttctttcatt tatacatcgt tttgccttct tttatgtaac
     2581 tatactcctc taagtttcaa tcttggccat gtaacctctg atctatagaa ttttttaaat
     2641 gactagaatt aatgcccatc ttttttttgg acctaaattc ttcatgaaaa tatattacga
     2701 gggcttattc agaagctttg gacttcttcg ccagaggttt ggtcaagtct ccaatcaagg
     2761 ttgtcggctt gtctaccttg ccagaaattt acgaaaagat ggaaaagggt caaatcgttg
     2821 gtagatacgt tgttgacact tctaaataag cgaatttctt atgatttatg atttttatta
     2881 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     2941 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     3001 agcatgaggt cgctcttatt gaccacacct ctaccggccg gtcgaaattc ccctacccta
     3061 tgaacatatt ccattttgta atttcgtgtc gtttctatta tgaatttcat ttataaagtt
     3121 tatgtacaaa tatcataaaa aaagagaatc tttttaagca aggattttct taacttcttc
     3181 ggcgacagca tcaccgactt cggtggtact gttggaacca cctaaatcac cagttctgat
     3241 acctgcatcc aaaacctttt taactgcatc ttcaatggcc ttaccttctt caggcaagtt
     3301 caatgacaat ttcaacatca ttgcagcaga caagatagtg gcgatagggt tgaccttatt
     3361 ctttggcaaa tctggagcag aaccgtggca tggttcgtac aaaccaaatg cggtgttctt
     3421 gtctggcaaa gaggccaagg acgcagatgg caacaaaccc aaggaacctg ggataacgga
     3481 ggcttcatcg gagatgatat caccaaacat gttgctggtg attataatac catttaggtg
     3541 ggttgggttc ttaactagga tcatggcggc agaatcaatc aattgatgtt gaaccttcaa
     3601 tgtaggaaat tcgttcttga tggtttcctc cacagttttt ctccataatc ttgaagaggc
     3661 caaaacatta gctttatcca aggaccaaat aggcaatggt ggctcatgtt gtagggccat
     3721 gaaagcggcc attcttgtga ttctttgcac ttctggaacg gtgtattgtt cactatccca
     3781 agcgacacca tcaccatcgt cttcctttct cttaccaaag taaatacctc ccactaattc
     3841 tctgacaaca acgaagtcag tacctttagc aaattgtggc ttgattggag ataagtctaa
     3901 aagagagtcg gatgcaaagt tacatggtct taagttggcg tacaattgaa gttctttacg
     3961 gatttttagt aaaccttgtt caggtctaac actacctgta ccccatttag gaccacccac
     4021 agcacctaac aaaacggcat caaccttctt ggaggcttcc agcgcctcat ctggaagtgg
     4081 gacacctgta gcgtcgatag cagcaccacc aattaaatga ttttcgaaat cgaacttgac
     4141 attggaacga acatcagaaa tagctttaag aaccttaatg gcttcggctg tgatttcttg
     4201 accaacgtgg tcacctggca aaacgacgat cttcttaggg gcagacatta gaatggtata
     4261 tccttgaaat atatatatat attgctgaaa tgtaaaaggt aagaaaagtt agaaagtaag
     4321 acgattgcta accacctatt ggaaaaaaca ataggtcctt aaataatatt gtcaacttca
     4381 agtattgtga tgcaagcatt tagtcatgaa cgcttctcta ttctatatga aaagccggtt
     4441 ccggcgctct cacctttcct ttttctccca atttttcagt tgaaaaaggt atatgcgtca
     4501 ggcgacctct gaaattaaca aaaaatttcc agtcatcgaa tttgattctg tgcgatagcg
     4561 cccctgtgtg ttctcgttat gttgaggaaa aaaataatgg ttgctaagag attcgaactc
     4621 ttgcatctta cgatacctga gtattcccac agttggggat ctcgactcta gctagaggat
     4681 caattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     4741 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     4801 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     4861 gctgataact tcgtataatg tatgctatac gaagttatta ggtctgaaga ggagtttacg
     4921 tccagccaag ctagcttggc tgcaggtcga gcggccgcga tccggaaccc ttaatataac
     4981 ttcgtataat gtatgctata cgaagttatc agctgcatta atgaatcggc caacgcgcgg
     5041 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     5101 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     5161 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     5221 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5281 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5341 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5401 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5461 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5521 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5581 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5641 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     5701 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5761 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5821 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5881 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5941 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     6001 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     6061 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     6121 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     6181 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     6241 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     6301 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6361 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6421 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6481 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6541 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6601 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6661 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6721 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6781 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6841 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6901 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6961 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     7021 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     7081 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     7141 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     7201 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cataacgcat
     7261 ttaagcataa acacgcacta tgccgttctt ctcatgtata tatatataca ggcaacacgc
     7321 agatataggt gcgacgtgaa cagtgagctg tatgtgcgca gctcgcgttg cattttcgga
     7381 agcgctcgtt ttcggaaacg ctttgaagtt cctattccga agttcctatt ctctagctag
     7441 aaagtatagg aacttcagag cgcttttgaa aaccaaaagc gctctgaaga cgcactttca
     7501 aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa tattgcgaat accgcttcca
     7561 caaacattgc tcaaaagtat ctctttgcta tatatctctg tgctatatcc ctatataacc
     7621 tacccatcca cctttcgctc cttgaacttg catctaaact cgacctctac attttttatg
     7681 tttatctcta gtattactct ttagacaaaa aaattgtagt aagaactatt catagagtga
     7741 atcgaaaaca atacgaaaat gtaaacattt cctatacgta gtatatagag acaaaataga
     7801 agaaaccgtt cataattttc tgaccaatga agaatcatca acgctatcac tttctgttca
     7861 caaagtatgc gcaatccaca tcggtataga atataatcgg ggatgccttt atcttgaaaa
     7921 aatgcacccg cagcttcgct agtaatcagt aaacgcggga agtggagtca ggcttttttt
     7981 atggaagaga aaatagacac caaagtagcc ttcttctaac cttaacggac ctacagtgca
     8041 aaaagttatc aagagactgc attatagagc gcacaaagga gaaaaaaagt aatctaagat
     8101 gctttgttag aaaaatagcg ctctcgggat gcatttttgt agaacaaaaa agaagtatag
     8161 attctttgtt ggtaaaatag cgctctcgcg ttgcatttct gttctgtaaa aatgcagctc
     8221 agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttttac aaaaatgaag
     8281 cacagattct tcgttggtaa aatagcgctt tcgcgttgca tttctgttct gtaaaaatgc
     8341 agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt tctacaaaat
     8401 gaagcacaga tgcttcgttg ct
//
