LOCUS       sequence-339513-        8815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes OptoEXP2 circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8815)
  AUTHORS   Zhao EM, Lalwani MA, Chen JM, Orillac P, Toettcher JE, Avalos JL
  TITLE     Optogenetic Amplification Circuits for Light-Induced Metabolic 
            Control.
  JOURNAL   ACS Synth Biol. 2021 May 21;10(5):1143-1154. doi: 
            10.1021/acssynbio.0c00642. Epub 2021 Apr 9.
  PUBMED    33835777
REFERENCE   2  (bases 1 to 8815)
  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: 
            "10.1021/acssynbio.0c00642"; journalName: "ACS Synth Biol"; date: 
            "2021-05-21- 21"; volume: "10"; issue: "5"; pages: "1143-1154"
COMMENT     Sequence Label: EZ-L545
FEATURES             Location/Qualifiers
     source          1..8815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(100..131)
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     terminator      complement(302..549)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     515..533
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(1438..1458)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(1488..1885)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     primer_bind     complement(2548..2567)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2549..2567)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2574..2591)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2574..2590)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2583..2605)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2731..3186
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2818..2837)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3028..3049
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3542..3561)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3661..3683
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3721..3739)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3823..3927
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3928..3996,3997..4788)
                     /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     complement(4146..4165)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4959..5547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5448..5467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5701..5718
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5835..5856
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5871..5876,5877..5894,5895..5901)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5909..5925
                     /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)."
     primer_bind     5914..5936
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5933..5949
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5933..5949
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5970..5988
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6314..6501
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     terminator      complement(7964..8129)
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     8050..8074
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     CDS             complement(join(8145..8815,1..37))
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI
                     CTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGNY
                     KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNF
                     KIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFV
                     TAAGITHGMDELYK"
     primer_bind     complement(8185..8205)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     join(8800..8815,1..13)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaattgggac aactccagtg aaaagttctt ctcctttgtc gacgctagcc atgattttag
       61 cggccgcggt ggctttacca acagtaccgg attgccaagc tggaagtcga gcttccatta
      121 tataccctct agtgtctctt aagcatggac taaaggctac ctataacgca tggactaaag
      181 gctacctata acgcatggac taaaggctac ctataacgca tggactaaag gctacctata
      241 acgcatggac taaaggctac ctactcgaag ctggcgaact cacgtacccg gcgataaggc
      301 cgcaaattaa agccttcgag cgtcccaaaa ccttctcaag caaggttttc agtataatgt
      361 tacatgcgta cacgcgtctg tacagaaaaa aaagaaaaat ttgaaatata aataacgttc
      421 ttaatactaa cataactata aaaaaataaa tagggaccta gacttcaggt tgtctaactc
      481 cttccttttc ggttagagcg gatgtggggg gagggcgtga atgtaagcgt gacataacta
      541 attacatgat cattactcga ggtcgactta aatacctgct tctacggcaa ttctaactaa
      601 atctgcactc gtttttaagt ttaatttttc cattaccaaa cctctgtgca ttttgacagt
      661 tttttctgaa agacccagtc tggcagccac ttctttattt cttaggcctg atgcaaccag
      721 agttgtgact tctaattgac gtggtgaaag agtttttaac atctccgcag cacgttctct
      781 acgggccatg cccatgttcg gttgatcgtc atccacttcc acttggctac caaggaagta
      841 tagtaattca tcatcgtcat catagattgg tgccaccaaa actgcattac gaaaaggagt
      901 accatccttt ttgtaattca aaatctcaac caacacgggt ttatgctcac gaacaccttg
      961 acggatctta tctgttagcc agggttcagt accagatccc tgtaagaacc tacaattacg
     1021 cccaacacat tcttcctcag aatacccagt tagatcggta aaggcctggt tgatagcaat
     1081 caaaggatta tcagccaatc tgggatcact aacaacactt gctatgggtg aggcctcgat
     1141 caaatccaat acccattggg ctggaggttg gacttcaact ctagtatcat ctgctccgaa
     1201 ttcacctcca tattcgtcga taccaagtgc atcagtaaac atctgttcaa attcgaaatc
     1261 tgccatgtcc aacgcgccat acggagcaga atcatgtggt gtaaatccgg ggccgggaga
     1321 atccccatca cctaacatgt ccagatcaaa atcatcaaga gcatcggcat gtgccatagc
     1381 tacatcctca ccgtctaaat gtaattcgtc accaagtgaa acgtccgttg gtggggctac
     1441 ttttctcttt tttttcgggc tagccatgat tttagcggcc gccaaaactt agattagatt
     1501 gctatgcttt ctttctaatg agcaagaagt aaaaaaagtt gtaatagaac aagaaaaatg
     1561 aaactgaaac ttgagaaatt gaagaccgtt tattaactta aatatcaatg ggaggtcatc
     1621 gaaagagaaa aaaatcaaaa aaaaaatttt caagaaaaag aaacgtgata aaaattttta
     1681 ttgccttttt cgacgaagaa aaagaaacga ggcggtctct tttttctttt ccaaaccttt
     1741 agtacgggta attaacgaca ccctagagga agaaagaggg gaaatttagt atgctgtgct
     1801 tgggtgtttt gaagtggtac ggcgatgcgc ggagtccgag aaaatctgga agagtaaaaa
     1861 aggagtagaa acattttgaa gctatcagct gtttcccggg agctggaact cccacctgca
     1921 acatgcggga tcctgagtaa ataagcggcc gccaccgcgg tggagctcag atcttgacac
     1981 cgattattta aagctgcagc atacgatata tatacatgtg tatatatgta tacctatgaa
     2041 tgtcagtaag tatgtatacg aacagtatga tactgaagat gacaaggtaa tgcatcattc
     2101 tatacgtgtc attctgaacg aggcgcgctt tccttttttc tttttgcttt ttcttttttt
     2161 ttctcttgaa ctccagaaaa aaaatataaa agagatggag gaacgggaaa aagttagttg
     2221 tggtgatagg tggcaagtgg tattccgtaa gaacaacaag aaaagcattt catattatgg
     2281 ctgaactgag cgaacaagtg caaaatttaa gcatcaacga caacaacgag aatggttatg
     2341 ttcctcctca cttaagagga aaaccaagaa gtgccagaaa taacagtagc aactacaata
     2401 acaacaacgg cggctacaac ggtggccgtg gcggtggcag cttctttagc aacaaccgtc
     2461 gtggtggtta cggcaacggt ggtttcttcg gtggaaacaa cggtggcagc ggccggccat
     2521 ttaaatgttt aaacggccac gctggccccc tatagtgagt cgtattacaa ttcactggcc
     2581 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     2641 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     2701 caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg
     2761 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2821 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2881 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2941 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3001 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3061 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3121 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     3181 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatagg
     3241 gtaataactg atataattaa attgaagctc taatttgtga gtttagtatt aggatctcta
     3301 cccttggcga aaagtcctct gccaacaatg atgatatctg atccaccact tacaacttcg
     3361 tcgagggttc tgtactgctg acccaatatg cgtatatata ccaatctaag tctgtgctcc
     3421 ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca aggaaaccga
     3481 aatcaaaaaa aagaataaaa aaaaaatgat gaattgaaaa ggtggtatgg tgcactctca
     3541 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     3601 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3661 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     3721 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3781 cacgcgccga ggacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3841 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3901 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     3961 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4021 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4081 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4141 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4201 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4261 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4321 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4381 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4441 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4501 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4561 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     4621 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     4681 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     4741 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     4801 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4861 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4921 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4981 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5041 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5101 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5161 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5221 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5281 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5341 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5401 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5461 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5521 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5581 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5641 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     5701 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     5761 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     5821 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     5881 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     5941 agctatgacc atgattacgc caagctcgga attaaccctc actaaaggga acaaaagctg
     6001 ggccggccat ttaaatgttt aaacggccac gctggccgct gcacggtcct gttccctagc
     6061 atgtacgtga gcgtatttcc ttttaaacca cgacgctttg tcttcattca acgtttccca
     6121 ttgttttttt ctactattgc tttgctgtgg gaaaaactta tcgaaagatg acgacttttt
     6181 cttaattctc gttttaagag cttggtgagc gctaggagtc actgccaggt atcgtttgaa
     6241 cacggcatta gtcagggaag tcataacaca gtcctttccc gcaattttct ttttctatta
     6301 ctcttggcct cctctagtac actctatatt tttttatgcc tcggtaatga ttttcatttt
     6361 tttttttcca cctagcggat gactcttttt ttttcttagc gattggcatt atcacataat
     6421 gaattataca ttatataaag taatgtgatt tcttcgaaga atatactaaa aaatgagcag
     6481 gcaagataaa cgaaggcaaa gaggctctat atgtatataa agctgcggat gccttctgat
     6541 ttaatctagt tctagtttac ttcaggttta cttaaagttg agataattga agcaaagaga
     6601 agcatcgagc aaaggatggc gtttgttaag agggttacgc aggagacgaa tatacagctg
     6661 gcgctggatc ttgacggtgg gtctgtttct gtacgggaga gcatactggg caaggaatat
     6721 gctagtggtg atgggcagac catccatgtg cacactggag ttgggttttt ggaccacatg
     6781 ttgactgcgc tggcgaagca tggcgggtgg tctctgatcc tggagtgtat aggggacttg
     6841 cacatcgatg accaccacac tgttgaggac tgtgggattg cgctgggcca agcgttcaag
     6901 gaggcgcttg gctccgtgcg tggtatcaag aggttcgggc atgggtttgc accactggac
     6961 gaggcgctga gccgcgctgt ggttgacttc tccaataggc ctttcgccgt ggtggagctg
     7021 ggcctgaagc gagaacgcat aggccagcta tccacagaga tgatcccgca cttcttggag
     7081 agtttcgcca ctgaggcgcg tatcaccatg catgtggact gtctgcgggg caccaacgac
     7141 caccaccgct ccgaatcagc tttcaaggcg ctcgccatcg ccatcagaga ggcaagaaca
     7201 cctacgggtc gcgatgacgt tccatccact aagggtgtcc tagcatagaa cacagcccac
     7261 agctaccacc atcaacaata tttatataga taacgtacac atagaaatca cacaaacaga
     7321 gtatttattc ttaactacat gaactaccat cagaccgtct gggccactat ataatgtgct
     7381 attcataaac gtgatcactc tacgtagcag gcgaccccag gtgaaaattt ttcagcgagc
     7441 tgccagattg tcaggtgaaa aactgaaaaa aacttctggg cgatgagctt gtggcgggaa
     7501 attaagtata taaagcagtt agtttcttct gcttcttgtg gttctgggta ttcttgtaat
     7561 atttggaata atagaaaacc aagactactg cctttctttt catattatag aggaagaaat
     7621 acgcacgaac acgatataga ggtaaatttt ttgcaaaaag ataccaaact ataaaagaaa
     7681 tagaagacta aactacaaaa aataaagaaa gtaatacata tatccaatat gactgagtta
     7741 gcagagcaag tttccatgcc tgaagggcaa caaagacgtg gtggttacgt tccacctcac
     7801 atccgtaacg gtggtggcag aagaggcgat tctgctcctc gtggtggttt caacaacggt
     7861 ggcggccgtg gtggtttctt cagaaacaac ggtgaccgtg gtggattcgg ctttgggtac
     7921 ctccggatat gttggcgcgc cttaatttat tcgccggcga taagagcgac ctcatgctat
     7981 acctgagaaa gcaacctgac ctacaggaaa gagttactca agaataagaa ttttcgtttt
     8041 aaaacctaag agtcacttta aaatttgtat acacttattt tttttataac ttatttaata
     8101 ataaaaatca taaatcataa gaaattcgct gatcattact cgagtttgta tagttcatcc
     8161 atgccatgtg taatcccagc agctgttaca aactcaagaa ggaccatgtg gtctctcttt
     8221 tcgttgggat ctttcgaaag ggcagattgt gtggacaggt aatggttgtc tggtaaaagg
     8281 acagggccat cgccaattgg agtattttgt tgataatggt ctgctagttg aacgcttcca
     8341 tcttcaatgt tgtgtctaat tttgaagtta actttgattc cattcttttg tttgtctgcc
     8401 atgatgtata cattgtgtga gttatagttg tattccaatt tgtgtccaag aatgtttcca
     8461 tcttctttaa aatcaatacc ttttaactcg attctattaa caagggtatc accttcaaac
     8521 ttgacttcag cacgtgtctt gtagttcccg tcatctttga aaaatatagt tctttcctgt
     8581 acataacctt cgggcatggc actcttgaaa aagtcatgcc gtttcatatg atctgggtat
     8641 cttgaaaagc attgaacacc ataagtgaaa gtagtgacaa gtgttggcca tggaacaggt
     8701 agttttccag tagtgcaaat aaatttaagg gtaagttttc cgtatgttgc atcaccttca
     8761 ccctctccac tgacagaaaa tttgtgccca ttaacatcac catctaattc aacaa
//
