LOCUS       sequence-343724-        8810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes OptoLAC1B circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8810)
  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 8810)
  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: pMAL658
FEATURES             Location/Qualifiers
     source          1..8810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(674..691)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      844..930
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1022..1049
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1068..1159
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1617..2072
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1704..1723)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1914..1935
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2628..3287)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3202..3221
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3288..3390)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3782..3799)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3916..4461)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3996..4015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4698..4745)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4741..4759
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4812..4829)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4870..5949)
                     /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     5911..5930
                     /label=LacI-R
                     /note="LacI, reverse primer"
     RBS             5956..5978
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      complement(6054..6081)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(6097..6168)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6216..6248)
                     /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"
     CDS             complement(6249..6959)
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             6966..6977
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        7526..7603
                     /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."
ORIGIN
        1 gctggattct gccggcttcg gcgtcacgct gtttgacgac gcgcaagcct ttctcgacgc
       61 cctgccgggt ctctccttcg gctgcgtcgt ctccgacgtg cgcatgccgg gccttgacgg
      121 catcgagctg ttgaagcgga tgaaggcgca gcaaagcccc tttccgatcc tcatcatgac
      181 cggtcacggc gacgtgccgc tcgcggtcga ggcgatgaag ttaggggcgg tggactttct
      241 ggaaaagcct ttcgaggacg accgcctcac cgccatgatc gaatcggcga tccgccaggc
      301 cgagccggcc gccaagagcg aggccgtcgc gcaggatatc gccgcccgcg tcgcctcgtt
      361 gagccccagg gagcgccagg tcatggaagg gctgatcgcc ggcctttcca acaagctgat
      421 cgcccgcgag tacgacatca gcccgcgcac catcgaggtg tatcgggcca acgtcatgac
      481 caagatgcag gccaacagcc tttcggagct ggttcgcctc gcgatgcgcg ccggcatgct
      541 caacgattga caattgatgt aagttagctc actcattagg caccgggatc tcgaccgatg
      601 cccttgagag ccttcaaccc agtctgcagg catgcaagct tggctgtttt ggcggatgag
      661 agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga
      721 atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga
      781 aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg
      841 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
      901 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
      961 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
     1021 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
     1081 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     1141 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     1201 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     1261 tgaagatcag ttgggtgcag caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttacgc
     1621 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     1681 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     1741 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     1801 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     1861 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     1921 cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg
     1981 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2041 gaattttaac aaaatattaa cgtttacaat ttaaaaggat ctaggtgaag atcctttttg
     2101 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     2161 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2221 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2281 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     2341 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2401 taatcctgtt accagtcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa
     2461 taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac
     2521 gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag
     2581 gcgtagcaac caggcgttta agggcaccaa taactgcctt aaaaaaatta cgccccgccc
     2641 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     2701 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     2761 tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     2821 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     2881 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     2941 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     3001 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     3061 gccatacgga attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     3121 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     3181 gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc
     3241 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     3301 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     3361 tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa gttggcccag
     3421 ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg atcttccgtc
     3481 acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttact gatttagtgt
     3541 atgatggtgt ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc
     3601 agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctagcggag
     3661 tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct tcatgtggca
     3721 ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata tattccgctt
     3781 cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg
     3841 aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa gtgagagggc
     3901 cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg aaatctgacg
     3961 ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4021 cggctccctc gtgcgctctc ctgttcctgc ctttcggttt accggtgtca ttccgctgtt
     4081 atggccgcgt ttgtctcatt ccacgcctga cactcagttc cgggtaggca gttcgctcca
     4141 agctggactg tatgcacgaa ccccccgttc agtccgaccg ctgcgcctta tccggtaact
     4201 atcgtcttga gtccaacccg gaaagacatg caaaagcacc actggcagca gccactggta
     4261 attgatttag aggagttagt cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca
     4321 agttttggtg actgcgctcc tccaagccag ttacctcggt tcaaagagtt ggtagctcag
     4381 agaaccttcg aaaaaccgcc ctgcaaggcg gttttttcgt tttcagagca agagattacg
     4441 cgcagaccaa aacgatctca agaagatcat cttattaatc agataaaata tttctaggct
     4501 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     4561 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg
     4621 ctcatgtttg acagcttatc atcgatgcat gcggccgccg tcccattcgc caatccggat
     4681 atagttcctc ctttcagcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     4741 gctagttatt gctcagcggt ggcagcagcc aactcagctt cctttcgggc tttgttagca
     4801 gccggatctc agtggtggtg gtggtggtgc tcattaagct gctaaagcgt agttttcgtc
     4861 gtttgctgcc tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4921 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     4981 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     5041 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     5101 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     5161 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     5221 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     5281 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     5341 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     5401 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     5461 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     5521 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     5581 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     5641 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     5701 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     5761 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     5821 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     5881 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     5941 tggtttcatg ctagctctcc ttcttaaagt taaacaaaat tatttctaga gcaaccatta
     6001 tcaccgccag aggtaaaata gtcaacacgc acggtgttac tctagtatat aaacgcagaa
     6061 aggcccaccc gaaggtgagc cagtgtgact ctagtagaga gcgttcaccg acaaacaaca
     6121 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcctggctct
     6181 agtagcgatc tacactagca ctatcagcgt tattaagcta ctaaagcgta gttttcgtcg
     6241 tttgcagcgc caaacgtctc ttcaggccac tgactagcga taactttccc cacaacggaa
     6301 caactctcat tgcatgggat cattgggtac tgtgggttta gtggttgtaa aaacacctga
     6361 ccgctatccc tgatcagttt cttgaaggta aactcatcac ccccaagtct ggctatgcag
     6421 aaatcacctg gctcaacagc ctgctcaggg tcaacgagaa ttaacattcc gtcaggaaag
     6481 ctcggcttgg agcctgttgg tgcggtcatg gaattacctt caacctcaag ccagaatgca
     6541 gaatcactgg cttttttggt tgtgcttacc catctctccg catcaccttt ggtaaaggtt
     6601 ctaagctcag gtgagaacat ccctgcctga acatgagaaa aaacagggta ctcatactca
     6661 cttctaagtg acggctgcat actaaccgct tcatacatct cgtagatttc tctggcgatt
     6721 gaagggctaa attcttcaac gctaactttg agaatttttg caagcaatgc ggcgttataa
     6781 gcatttaatg cattgatgcc attaaataaa gcaccaacgc ctgactgccc catccccatc
     6841 ttgtctgcga cagattcctg ggataagcca agttcatttt tctttttttc ataaattgct
     6901 ttaaggcgac gtgcgtcctc aagctgctct tgtgttaatg gtttcttttt tgtgctcatc
     6961 tagtatttct cctctttact agtactccgt tgtgatgacg cattggtacg cggtatcggg
     7021 aggttcgaaa atttcgagcg atatcttaag gggggtgcct tacgtagaac cccgtaggtc
     7081 atgcccgagg ccggtcctgg atggcgcggc ggatacgctt gagcaggttt tcgtcgagaa
     7141 gcggcttcaa aaccacgtct tttacgccgg cctcggcggc ccgggtcgag atgttttcgt
     7201 ccggatagcc ggtgatcagg atcacgggcg tagatctcga tcctctacgc cggacgcatc
     7261 gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc
     7321 gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg
     7381 gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt
     7441 gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg
     7501 cataagggag agcgtcgaga tcccggacac catcgaatgg tgcaaaacct ttcgcggtat
     7561 ggcatgatag cgcccggaag agagtcaatt gagggtggtg aatgtggcta gttttcaatc
     7621 atttgggata ccaggacagc tggaagtcat caaaaaagca cttgatcacg tgcgagtcgg
     7681 tgtggtaatt acagatcccg cacttgaaga taatcctatt gtctacgtaa atcaaggctt
     7741 tgttcaaatg accggctacg agaccgagga aattttagga aagaactgtc gcttcttaca
     7801 ggggaaacac acagatcctg cagaagtgga caacatcaga accgctttac aaaataaaga
     7861 accggtcacc gttcagatcc aaaactacaa aaaagacgga acgatgttct ggaatgaatt
     7921 aaatattgat ccaatggaaa tagaggataa aacgtatttt gtcggaattc agaatgatat
     7981 caccgagcac cagcagaccc aggcgcgcct ccaggaactg caatccgagc tcgtccacgt
     8041 ctccaggctg agcgccatgg gcgaaatggc gtccgcgctc gcgcacgagc tcaaccagcc
     8101 gctggcggcg atcagcaact acatgaaggg ctcgcggcgg ctgcttgccg gcagcagtga
     8161 tccgaacaca ccgaaggtcg aaagcgccct ggaccgcgcc gccgagcagg cgctgcgcgc
     8221 cggccagatc atccggcgcc tgcgcgactt cgttgcccgc ggcgaatcgg agaagcgggt
     8281 cgagagtctc tccaagctga tcgaggaggc cggcgcgctc gggcttgccg gcgcgcgcga
     8341 gcagaacgtg cagctccgct tcagtctcga tccgggcgcc gatctcgttc tcgccgaccg
     8401 ggtgcagatc cagcaggtcc tggtcaacct gttccgcaac gcgctggaag cgatggctca
     8461 gtcgcagcga cgcgagctcg tcgtcaccaa cacccccgcc gccgacgaca tgatcgaggt
     8521 cgaagtgtcc gacaccggca gcggtttcca ggacgacgtc attccgaacc tgtttcagac
     8581 tttcttcacc accaaggaca ccggcatggg cgtgggactg tccatcagcc gctcgatcat
     8641 cgaagctcac ggcgggcgca tgtgggccga gagcaacgca tcgggcgggg cgaccttccg
     8701 cttcaccctc ccggcagccg acgagatgat aggaggtcta gcatgacgac caagggacat
     8761 atctacgtca tcgacgacga cgcggcgatg cgggattcgc tgaatttcct
//
