LOCUS       Exported                8923 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pGal4AD-CIB1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8923)
  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 Oct 31. ():.
  PUBMED    21037589
REFERENCE   2  (bases 1 to 8923)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8923
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             1948..2457
                     /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      3351..3538
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3406..3430)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(3655..4749)
                     /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(4750..5155)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(5197..5213)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5197..5213)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5210..5232)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5221..5237
                     /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(5245..5275)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5290..5311
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(5366..5399)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(5477..5510)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(5579..5596)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5750..6338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5830..5849)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6509..7369)
                     /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     7132..7151
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7370..7474)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7542..7560
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7598..7620)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7720..7739
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      7756..8920
                     /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 ccgaagctag cgccaccatg aatggagcta taggaggtga ccttttgctc
     1981 aattttcctg acatgtcggt cctagagcgc caaagggctc acctcaagta cctcaatccc
     2041 acctttgatt ctcctctcgc cggcttcttt gccgattctt caatgattac cggcggcgag
     2101 atggacagct atctttcgac tgccggtttg aatcttccga tgatgtacgg tgagacgacg
     2161 gtggaaggtg attcaagact ctcaatttcg ccggaaacga cgcttgggac tggaaatttc
     2221 aagaaacgga agtttgatac agagactaag gattgtaatg agaagaagaa gaagatgacg
     2281 atgaacagag atgacctagt agaagaagga gaagaagaga agtcgaaaat aacagagcaa
     2341 aacaatggga gcacaaaaag catcaagaag atgaaacaca aagccaagaa agaagagaac
     2401 aatttctcta atgattcatc taaagtgacg aaggaattgg agaaaacgga ttatattcat
     2461 gttcgtgcac gacgaggcca agccactgat agtcacagca tagcagaacg agttagaaga
     2521 gaaaagatca gtgagagaat gaagtttcta caagatttgg ttcctggatg cgacaagatc
     2581 acaggcaaag cagggatgct tgatgaaatc attaactatg ttcagtctct tcagagacaa
     2641 atcgagttct tatcgatgaa actagcaatt gtgaatccaa ggccggattt tgatatggat
     2701 gacatttttg ccaaagaggt tgcctcaact ccaatgactg tggtgccatc tcctgaaatg
     2761 gttctttccg gttattctca tgagatggtt cactctggtt attctagtga gatggttaac
     2821 tccggttacc ttcatgtcaa tccaatgcag caagtgaata ccagttctga tccattgtca
     2881 tgcttcaaca atggcgaagc tccttcgatg tgggactctc atgtgcagaa tctctatggc
     2941 aatttaggag taccggtcat cgagctcgag ctgcagatga atcgtagata ctgaaaaacc
     3001 ccgcaagttc acttcaactg tgcatcgtgc accatctcaa tttctttcat ttatacatcg
     3061 ttttgccttc ttttatgtaa ctatactcct ctaagtttca atcttggcca tgtaacctct
     3121 gatctataga attttttaaa tgactagaat taatgcccat cttttttttg gacctaaatt
     3181 cttcatgaaa atatattacg agggcttatt cagaagcttt ggacttcttc gccagaggtt
     3241 tggtcaagtc tccaatcaag gttgtcggct tgtctacctt gccagaaatt tacgaaaaga
     3301 tggaaaaggg tcaaatcgtt ggtagatacg ttgttgacac ttctaaataa gcgaatttct
     3361 tatgatttat gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt
     3421 aaagtgactc ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg
     3481 tcaggttgct ttctcaggta tagcatgagg tcgctcttat tgaccacacc tctaccggcc
     3541 ggtcgaaatt cccctaccct atgaacatat tccattttgt aatttcgtgt cgtttctatt
     3601 atgaatttca tttataaagt ttatgtacaa atatcataaa aaaagagaat ctttttaagc
     3661 aaggattttc ttaacttctt cggcgacagc atcaccgact tcggtggtac tgttggaacc
     3721 acctaaatca ccagttctga tacctgcatc caaaaccttt ttaactgcat cttcaatggc
     3781 cttaccttct tcaggcaagt tcaatgacaa tttcaacatc attgcagcag acaagatagt
     3841 ggcgataggg ttgaccttat tctttggcaa atctggagca gaaccgtggc atggttcgta
     3901 caaaccaaat gcggtgttct tgtctggcaa agaggccaag gacgcagatg gcaacaaacc
     3961 caaggaacct gggataacgg aggcttcatc ggagatgata tcaccaaaca tgttgctggt
     4021 gattataata ccatttaggt gggttgggtt cttaactagg atcatggcgg cagaatcaat
     4081 caattgatgt tgaaccttca atgtaggaaa ttcgttcttg atggtttcct ccacagtttt
     4141 tctccataat cttgaagagg ccaaaacatt agctttatcc aaggaccaaa taggcaatgg
     4201 tggctcatgt tgtagggcca tgaaagcggc cattcttgtg attctttgca cttctggaac
     4261 ggtgtattgt tcactatccc aagcgacacc atcaccatcg tcttcctttc tcttaccaaa
     4321 gtaaatacct cccactaatt ctctgacaac aacgaagtca gtacctttag caaattgtgg
     4381 cttgattgga gataagtcta aaagagagtc ggatgcaaag ttacatggtc ttaagttggc
     4441 gtacaattga agttctttac ggatttttag taaaccttgt tcaggtctaa cactacctgt
     4501 accccattta ggaccaccca cagcacctaa caaaacggca tcaaccttct tggaggcttc
     4561 cagcgcctca tctggaagtg ggacacctgt agcgtcgata gcagcaccac caattaaatg
     4621 attttcgaaa tcgaacttga cattggaacg aacatcagaa atagctttaa gaaccttaat
     4681 ggcttcggct gtgatttctt gaccaacgtg gtcacctggc aaaacgacga tcttcttagg
     4741 ggcagacatt agaatggtat atccttgaaa tatatatata tattgctgaa atgtaaaagg
     4801 taagaaaagt tagaaagtaa gacgattgct aaccacctat tggaaaaaac aataggtcct
     4861 taaataatat tgtcaacttc aagtattgtg atgcaagcat ttagtcatga acgcttctct
     4921 attctatatg aaaagccggt tccggcgctc tcacctttcc tttttctccc aatttttcag
     4981 ttgaaaaagg tatatgcgtc aggcgacctc tgaaattaac aaaaaatttc cagtcatcga
     5041 atttgattct gtgcgatagc gcccctgtgt gttctcgtta tgttgaggaa aaaaataatg
     5101 gttgctaaga gattcgaact cttgcatctt acgatacctg agtattccca cagttgggga
     5161 tctcgactct agctagagga tcaattcgta atcatggtca tagctgtttc ctgtgtgaaa
     5221 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5281 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5341 gtcgggaaac ctgtcgtgcc agctgataac ttcgtataat gtatgctata cgaagttatt
     5401 aggtctgaag aggagtttac gtccagccaa gctagcttgg ctgcaggtcg agcggccgcg
     5461 atccggaacc cttaatataa cttcgtataa tgtatgctat acgaagttat cagctgcatt
     5521 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5581 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5641 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5701 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5761 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5821 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5881 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5941 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6001 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6061 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6121 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6181 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6241 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6301 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6361 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6421 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6481 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6541 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6601 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6661 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6721 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6781 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6841 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6901 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6961 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     7021 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     7081 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     7141 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     7201 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     7261 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7321 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7381 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7441 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7501 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     7561 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     7621 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     7681 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     7741 tgagagtgca ccataacgca tttaagcata aacacgcact atgccgttct tctcatgtat
     7801 atatatatac aggcaacacg cagatatagg tgcgacgtga acagtgagct gtatgtgcgc
     7861 agctcgcgtt gcattttcgg aagcgctcgt tttcggaaac gctttgaagt tcctattccg
     7921 aagttcctat tctctagcta gaaagtatag gaacttcaga gcgcttttga aaaccaaaag
     7981 cgctctgaag acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa
     8041 atattgcgaa taccgcttcc acaaacattg ctcaaaagta tctctttgct atatatctct
     8101 gtgctatatc cctatataac ctacccatcc acctttcgct ccttgaactt gcatctaaac
     8161 tcgacctcta cattttttat gtttatctct agtattactc tttagacaaa aaaattgtag
     8221 taagaactat tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt
     8281 agtatataga gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc
     8341 aacgctatca ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg
     8401 gggatgcctt tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg
     8461 aagtggagtc aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa
     8521 ccttaacgga cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg
     8581 agaaaaaaag taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg
     8641 tagaacaaaa aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc
     8701 tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat
     8761 ttttgtttta caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc
     8821 atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt
     8881 tgcatttttg ttctacaaaa tgaagcacag atgcttcgtt gct
//
