LOCUS       sequence-340149-        7650 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes OptoLAC3 circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7650)
  AUTHORS   Lalwani MA, Ip SS, Carrasco-Lopez C, Day C, Zhao EM, Kawabe H, 
            Avalos JL
  TITLE     Optogenetic control of the lac operon for bacterial chemical and 
            protein production.
  JOURNAL   Nat Chem Biol. 2021 Jan;17(1):71-79. doi: 10.1038/s41589-020-0639-1.
            Epub 2020 Sep 7.
  PUBMED    32895498
REFERENCE   2  (bases 1 to 7650)
  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/s41589-020-0639-1"; journalName: "Nat Chem Biol"; date: 
            "2021-01"; volume: "17"; issue: "1"; pages: "71-79"
COMMENT     Sequence Label: pMAL630
FEATURES             Location/Qualifiers
     source          1..7650
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             516..533
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(586..604)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      600..647
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             771..962
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(956..978)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1064..1203
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1119..1138
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1218..1235)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1389..1977)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1469..1488)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2088..2107)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2099..2914
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2099..2914
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2172..2191)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3006..3461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3143..3164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3355..3374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3577..3596)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3802..3879
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     protein_bind    5995..6011
                     /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)."
     RBS             6018..6029
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6036..6746
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             6747..6779
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     terminator      6827..6898
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      6914..6941
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             7017..7039
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(7046..7650,1..475)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(7065..7084)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
       61 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
      121 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
      181 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
      241 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
      301 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
      361 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
      421 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcaggcagc
      481 aaacgacgaa aactacgctg cagcagttta atgagcacca ccaccaccac cactgagatc
      541 cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac
      601 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
      661 ctatatccgg attggcggat ccatcagtaa cccgtatcgt gagcatcctc tctcgtttca
      721 tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac
      781 aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg
      841 agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc
      901 acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
      961 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     1021 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     1081 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     1141 gagagtgcac cattgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     1201 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1261 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1321 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1381 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1441 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1501 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1561 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1621 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1681 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1741 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1801 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     1861 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     1921 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1981 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2041 attttggtca tgaacaataa aactgtctgc ttacataaac agtaatacaa ggggtgttat
     2101 gagccatatt caacgggaaa cgtcttgctc taggccgcga ttaaattcca acatggatgc
     2161 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
     2221 tcgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
     2281 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
     2341 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
     2401 ccccgggaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
     2461 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
     2521 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
     2581 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
     2641 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
     2701 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
     2761 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     2821 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     2881 gcagtttcat ttgatgctcg atgagttttt ctaagaatta attcatgagc ggatacatat
     2941 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3001 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3061 ctcatttttt aaccaataga ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc caatccggat
     3481 atagttcctc ctttcagcaa aaaacccaga tctcgatcct ctacgccgga cgcatcgtgg
     3541 ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg
     3601 gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg
     3661 caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg
     3721 cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata
     3781 agggagagcg tcgagatccc ggacaccatc gaatggtgca aaacctttcg cggtatggca
     3841 tgatagcgcc cggaagagag tcaattgagg gtggtgaatg tggctagttt tcaatcattt
     3901 gggataccag gacagctgga agtcatcaaa aaagcacttg atcacgtgcg agtcggtgtg
     3961 gtaattacag atcccgcact tgaagataat cctattgtct acgtaaatca aggctttgtt
     4021 caaatgaccg gctacgagac cgaggaaatt ttaggaaaga actgtcgctt cttacagggg
     4081 aaacacacag atcctgcaga agtggacaac atcagaaccg ctttacaaaa taaagaaccg
     4141 gtcaccgttc agatccaaaa ctacaaaaaa gacggaacga tgttctggaa tgaattaaat
     4201 attgatccaa tggaaataga ggataaaacg tattttgtcg gaattcagaa tgatatcacc
     4261 gagcaccagc agacccaggc gcgcctccag gaactgcaat ccgagctcgt ccacgtctcc
     4321 aggctgagcg ccatgggcga aatggcgtcc gcgctcgcgc acgagctcaa ccagccgctg
     4381 gcggcgatca gcaactacat gaagggctcg cggcggctgc ttgccggcag cagtgatccg
     4441 aacacaccga aggtcgaaag cgccctggac cgcgccgccg agcaggcgct gcgcgccggc
     4501 cagatcatcc ggcgcctgcg cgacttcgtt gcccgcggcg aatcggagaa gcgggtcgag
     4561 agtctctcca agctgatcga ggaggccggc gcgctcgggc ttgccggcgc gcgcgagcag
     4621 aacgtgcagc tccgcttcag tctcgatccg ggcgccgatc tcgttctcgc cgaccgggtg
     4681 cagatccagc aggtcctggt caacctgttc cgcaacgcgc tggaagcgat ggctcagtcg
     4741 cagcgacgcg agctcgtcgt caccaacacc cccgccgccg acgacatgat cgaggtcgaa
     4801 gtgtccgaca ccggcagcgg tttccaggac gacgtcattc cgaacctgtt tcagactttc
     4861 ttcaccacca aggacaccgg catgggcgtg ggactgtcca tcagccgctc gatcatcgaa
     4921 gctcacggcg ggcgcatgtg ggccgagagc aacgcatcgg gcggggcgac cttccgcttc
     4981 accctcccgg cagccgacga gatgatagga ggtctagcat gacgaccaag ggacatatct
     5041 acgtcatcga cgacgacgcg gcgatgcggg attcgctgaa tttcctgctg gattctgccg
     5101 gcttcggcgt cacgctgttt gacgacgcgc aagcctttct cgacgccctg ccgggtctct
     5161 ccttcggctg cgtcgtctcc gacgtgcgca tgccgggcct tgacggcatc gagctgttga
     5221 agcggatgaa ggcgcagcaa agcccctttc cgatcctcat catgaccggt cacggcgacg
     5281 tgccgctcgc ggtcgaggcg atgaagttag gggcggtgga ctttctggaa aagcctttcg
     5341 aggacgaccg cctcaccgcc atgatcgaat cggcgatccg ccaggccgag ccggccgcca
     5401 agagcgaggc cgtcgcgcag gatatcgccg cccgcgtcgc ctcgttgagc cccagggagc
     5461 gccaggtcat ggaagggctg atcgccggcc tttccaacaa gctgatcgcc cgcgagtacg
     5521 acatcagccc gcgcaccatc gaggtgtatc gggccaacgt catgaccaag atgcaggcca
     5581 acagcctttc ggagctggtt cgcctcgcga tgcgcgccgg catgctcaac gattgacaat
     5641 tgatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt
     5701 caacccagtc tgtacgcggc cgcgtccggc gtagaggatc gagatctacg cccgtgatcc
     5761 tgatcaccgg ctatccggac gaaaacatct cgacccgggc cgccgaggcc ggcgtaaaag
     5821 acgtggtttt gaagccgctt ctcgacgaaa acctgctcaa gcgtatccgc cgcgccatcc
     5881 aggaccggcc tcgggcatga cctacggggt tctacgtaag gcacccccct taagatatcg
     5941 ctcgaaattt tcgaacctcc cgataccgcg taccaatgcg tcatcacaac ggagttgtga
     6001 gcggataaca aactagtaaa gaggagaaat actagatgag cacaaaaaag aaaccattaa
     6061 cacaagagca gcttgaggac gcacgtcgcc ttaaagcaat ttatgaaaaa aagaaaaatg
     6121 aacttggctt atcccaggaa tctgtcgcag acaagatggg gatggggcag tcaggcgttg
     6181 gtgctttatt taatggcatc aatgcattaa atgcttataa cgccgcattg cttgcaaaaa
     6241 ttctcaaagt tagcgttgaa gaatttagcc cttcaatcgc cagagaaatc tacgagatgt
     6301 atgaagcggt tagtatgcag ccgtcactta gaagtgagta tgagtaccct gttttttctc
     6361 atgttcaggc agggatgttc tcacctgagc ttagaacctt taccaaaggt gatgcggaga
     6421 gatgggtaag cacaaccaaa aaagccagtg attctgcatt ctggcttgag gttgaaggta
     6481 attccatgac cgcaccaaca ggctccaagc cgagctttcc tgacggaatg ttaattctcg
     6541 ttgaccctga gcaggctgtt gagccaggtg atttctgcat agccagactt gggggtgatg
     6601 agtttacctt caagaaactg atcagggata gcggtcaggt gtttttacaa ccactaaacc
     6661 cacagtaccc aatgatccca tgcaatgaga gttgttccgt tgtggggaaa gttatcgcta
     6721 gtcagtggcc tgaagagacg tttggcgctg caaacgacga aaactacgct ttagtagctt
     6781 aataacgctg atagtgctag tgtagatcgc tactagagcc aggcatcaaa taaaacgaaa
     6841 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcta
     6901 ctagagtcac actggctcac cttcgggtgg gcctttctgc gtttatatac tagagtaaca
     6961 ccgtgcgtgt tgactatttt acctctggcg gtgataatgg ttgctctaga aataattttg
     7021 tttaacttta agaaggagag ctagcatgaa accagtaacg ttatacgatg tcgcagagta
     7081 tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc
     7141 gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt
     7201 ggcacaacaa ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc
     7261 cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc
     7321 cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca
     7381 caatcttctc gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga
     7441 tgccattgct gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga
     7501 ccagacaccc atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga
     7561 gcatctggtc gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt
     7621 ctcggcgcgt ctgcgtctgg ctggctggca
//
