LOCUS       sequence-340148-        8190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes OptoLAC1 circuit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8190)
  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 8190)
  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: pMAL294
FEATURES             Location/Qualifiers
     source          1..8190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1016..1093)
                     /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."
     primer_bind     1299..1318
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             1642..1653
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1660..2370
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             2371..2403
                     /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      2451..2522
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2538..2565
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             2641..2663
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2670..3749
                     /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(2689..2708)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             3790..3807
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3860..3878)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3874..3921
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3958..4413
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4045..4064)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4255..4276
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4505..5320)
                     /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             complement(4505..5320)
                     /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     5228..5247
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5312..5331
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5442..6030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5931..5950
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6184..6201
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    6216..6355
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(6281..6300)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6441..6463
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6457..6648)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gtcccacgcc catgccggtg tccttggtgg tgaagaaagt ctgaaacagg ttcggaatga
       61 cgtcgtcctg gaaaccgctg ccggtgtcgg acacttcgac ctcgatcatg tcgtcggcgg
      121 cgggggtgtt ggtgacgacg agctcgcgtc gctgcgactg agccatcgct tccagcgcgt
      181 tgcggaacag gttgaccagg acctgctgga tctgcacccg gtcggcgaga acgagatcgg
      241 cgcccggatc gagactgaag cggagctgca cgttctgctc gcgcgcgccg gcaagcccga
      301 gcgcgccggc ctcctcgatc agcttggaga gactctcgac ccgcttctcc gattcgccgc
      361 gggcaacgaa gtcgcgcagg cgccggatga tctggccggc gcgcagcgcc tgctcggcgg
      421 cgcggtccag ggcgctttcg accttcggtg tgttcggatc actgctgccg gcaagcagcc
      481 gccgcgagcc cttcatgtag ttgctgatcg ccgccagcgg ctggttgagc tcgtgcgcga
      541 gcgcggacgc catttcgccc atggcgctca gcctggagac gtggacgagc tcggattgca
      601 gttcctggag gcgcgcctgg gtctgctggt gctcggtgat atcattctga attccgacaa
      661 aatacgtttt atcctctatt tccattggat caatatttaa ttcattccag aacatcgttc
      721 cgtctttttt gtagttttgg atctgaacgg tgaccggttc tttattttgt aaagcggttc
      781 tgatgttgtc cacttctgca ggatctgtgt gtttcccctg taagaagcga cagttctttc
      841 ctaaaatttc ctcggtctcg tagccggtca tttgaacaaa gccttgattt acgtagacaa
      901 taggattatc ttcaagtgcg ggatctgtaa ttaccacacc gactcgcacg tgatcaagtg
      961 cttttttgat gacttccagc tgtcctggta tcccaaatga ttgaaaacta gccacattca
     1021 ccaccctcaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcacc
     1081 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     1141 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     1201 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     1261 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     1321 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     1381 gagatctacg cccgtgatcc tgatcaccgg ctatccggac gaaaacatct cgacccgggc
     1441 cgccgaggcc ggcgtaaaag acgtggtttt gaagccgctt ctcgacgaaa acctgctcaa
     1501 gcgtatccgc cgcgccatcc aggaccggcc tcgggcatga cctacggggt tctacgtaag
     1561 gcacccccct taagatatcg ctcgaaattt tcgaacctcc cgataccgcg taccaatgcg
     1621 tcatcacaac ggagtactag taaagaggag aaatactaga tgagcacaaa aaagaaacca
     1681 ttaacacaag agcagcttga ggacgcacgt cgccttaaag caatttatga aaaaaagaaa
     1741 aatgaacttg gcttatccca ggaatctgtc gcagacaaga tggggatggg gcagtcaggc
     1801 gttggtgctt tatttaatgg catcaatgca ttaaatgctt ataacgccgc attgcttgca
     1861 aaaattctca aagttagcgt tgaagaattt agcccttcaa tcgccagaga aatctacgag
     1921 atgtatgaag cggttagtat gcagccgtca cttagaagtg agtatgagta ccctgttttt
     1981 tctcatgttc aggcagggat gttctcacct gagcttagaa cctttaccaa aggtgatgcg
     2041 gagagatggg taagcacaac caaaaaagcc agtgattctg cattctggct tgaggttgaa
     2101 ggtaattcca tgaccgcacc aacaggctcc aagccgagct ttcctgacgg aatgttaatt
     2161 ctcgttgacc ctgagcaggc tgttgagcca ggtgatttct gcatagccag acttgggggt
     2221 gatgagttta ccttcaagaa actgatcagg gatagcggtc aggtgttttt acaaccacta
     2281 aacccacagt acccaatgat cccatgcaat gagagttgtt ccgttgtggg gaaagttatc
     2341 gctagtcagt ggcctgaaga gacgtttggc gctgcaaacg acgaaaacta cgctttagta
     2401 gcttaataac gctgatagtg ctagtgtaga tcgctactag agccaggcat caaataaaac
     2461 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     2521 tctactagag tcacactggc tcaccttcgg gtgggccttt ctgcgtttat atactagagt
     2581 aacaccgtgc gtgttgacta ttttacctct ggcggtgata atggttgctc tagaaataat
     2641 tttgtttaac tttaagaagg agagctagca tgaaaccagt aacgttatac gatgtcgcag
     2701 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
     2761 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
     2821 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
     2881 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
     2941 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
     3001 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc
     3061 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
     3121 ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg
     3181 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
     3241 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc
     3301 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc
     3361 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc
     3421 tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat atctcggtag
     3481 tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc accatcaaac
     3541 aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc tctcagggcc
     3601 aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa accaccctgg
     3661 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     3721 gacaggtttc ccgactggaa agcgggcagg cagcaaacga cgaaaactac gctttagcag
     3781 cttaatgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
     3841 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     3901 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc gaatgggacg
     3961 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4021 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4081 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4141 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4201 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4261 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4321 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4381 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     4441 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgaatta
     4501 attcttagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     4561 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     4621 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     4681 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     4741 cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc tttccagact tgttcaacag
     4801 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     4861 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     4921 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     4981 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     5041 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     5101 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     5161 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     5221 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     5281 gcggcctaga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     5341 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     5401 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5461 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5521 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5581 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5641 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5701 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5761 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5821 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5881 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5941 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6001 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6061 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6121 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6181 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     6241 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
     6301 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
     6361 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
     6421 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
     6481 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
     6541 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
     6601 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
     6661 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
     6721 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
     6781 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
     6841 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
     6901 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
     6961 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
     7021 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
     7081 cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg cgataatggc
     7141 ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg
     7201 caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc
     7261 ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa
     7321 gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg
     7381 gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac
     7441 atcaattgtc aatcgttgag catgccggcg cgcatcgcga ggcgaaccag ctccgaaagg
     7501 ctgttggcct gcatcttggt catgacgttg gcccgataca cctcgatggt gcgcgggctg
     7561 atgtcgtact cgcgggcgat cagcttgttg gaaaggccgg cgatcagccc ttccatgacc
     7621 tggcgctccc tggggctcaa cgaggcgacg cgggcggcga tatcctgcgc gacggcctcg
     7681 ctcttggcgg ccggctcggc ctggcggatc gccgattcga tcatggcggt gaggcggtcg
     7741 tcctcgaaag gcttttccag aaagtccacc gcccctaact tcatcgcctc gaccgcgagc
     7801 ggcacgtcgc cgtgaccggt catgatgagg atcggaaagg ggctttgctg cgccttcatc
     7861 cgcttcaaca gctcgatgcc gtcaaggccc ggcatgcgca cgtcggagac gacgcagccg
     7921 aaggagagac ccggcagggc gtcgagaaag gcttgcgcgt cgtcaaacag cgtgacgccg
     7981 aagccggcag aatccagcag gaaattcagc gaatcccgca tcgccgcgtc gtcgtcgatg
     8041 acgtagatat gtcccttggt cgtcatgcta gacctcctat catctcgtcg gctgccggga
     8101 gggtgaagcg gaaggtcgcc ccgcccgatg cgttgctctc ggcccacatg cgcccgccgt
     8161 gagcttcgat gatcgagcgg ctgatggaca
//
