LOCUS       sequence-340112-        8190 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes OptoLAC2 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: pMAL296
FEATURES             Location/Qualifiers
     source          1..8190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             468..485
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(538..556)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      552..599
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      636..1091
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(723..742)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     933..954
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1183..1998)
                     /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(1183..1998)
                     /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     1906..1925
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1990..2009
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2120..2708
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2609..2628
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2862..2879
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2894..3033
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2959..2978)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3119..3141
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3135..3326)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     promoter        complement(5884..5961)
                     /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     6167..6186
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     RBS             6510..6521
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             6528..7238
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     CDS             7239..7271
                     /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      7319..7390
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      7406..7433
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             7509..7531
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             join(7538..8190,1..427)
                     /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(7557..7576)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
       61 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
      121 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
      181 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
      241 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
      301 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
      361 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      421 cgggcaggca gcaaacgacg aaaactacgc tgcagcagtt taatgagcac caccaccacc
      481 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
      541 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
      601 tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag
      661 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
      721 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
      781 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
      841 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
      901 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
      961 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggtcta
     1021 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     1081 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1141 ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa actcatcgag
     1201 catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt tttgaaaaag
     1261 ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg caagatcctg
     1321 gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt tcccctcgtc
     1381 aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg gtgagaatgg
     1441 caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac gctcgtcatc
     1501 aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa
     1561 tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa
     1621 cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta atacctggaa
     1681 tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag tacggataaa
     1741 atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga ccatctcatc
     1801 tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg gcgcatcggg
     1861 cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc gagcccattt
     1921 atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc aagacgtttc
     1981 ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag acagttttat
     2041 tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2101 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2161 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     2221 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     2281 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     2341 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2401 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2461 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2521 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2581 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2641 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2701 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2761 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     2821 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     2881 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2941 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     3001 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     3061 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     3121 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     3181 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     3241 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     3301 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     3361 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     3421 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     3481 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     3541 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     3601 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     3661 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     3721 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     3781 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     3841 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     3901 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     3961 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     4021 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     4081 ggtcgagatc ccggtgccta atgagtgagc taacttacat caattgtcaa tcgttgagca
     4141 tgccggcgcg catcgcgagg cgaaccagct ccgaaaggct gttggcctgc atcttggtca
     4201 tgacgttggc ccgatacacc tcgatggtgc gcgggctgat gtcgtactcg cgggcgatca
     4261 gcttgttgga aaggccggcg atcagccctt ccatgacctg gcgctccctg gggctcaacg
     4321 aggcgacgcg ggcggcgata tcctgcgcga cggcctcgct cttggcggcc ggctcggcct
     4381 ggcggatcgc cgattcgatc atggcggtga ggcggtcgtc ctcgaaaggc ttttccagaa
     4441 agtccaccgc ccctaacttc atcgcctcga ccgcgagcgg cacgtcgccg tgaccggtca
     4501 tgatgaggat cggaaagggg ctttgctgcg ccttcatccg cttcaacagc tcgatgccgt
     4561 caaggcccgg catgcgcacg tcggagacga cgcagccgaa ggagagaccc ggcagggcgt
     4621 cgagaaaggc ttgcgcgtcg tcaaacagcg tgacgccgaa gccggcagaa tccagcagga
     4681 aattcagcga atcccgcatc gccgcgtcgt cgtcgatgac gtagatatgt cccttggtcg
     4741 tcatgctaga cctcctatca tctcgtcggc tgccgggagg gtgaagcgga aggtcgcccc
     4801 gcccgatgcg ttgctctcgg cccacatgcg cccgccgtga gcttcgatga tcgagcggct
     4861 gatggacagt cccacgccca tgccggtgtc cttggtggtg aagaaagtct gaaacaggtt
     4921 cggaatgacg tcgtcctgga aaccgctgcc ggtgtcggac acttcgacct cgatcatgtc
     4981 gtcggcggcg ggggtgttgg tgacgacgag ctcgcgtcgc tgcgactgag ccatcgcttc
     5041 cagcgcgttg cggaacaggt tgaccaggac ctgctggatc tgcacccggt cggcgagaac
     5101 gagatcggcg cccggatcga gactgaagcg gagctgcacg ttctgctcgc gcgcgccggc
     5161 aagcccgagc gcgccggcct cctcgatcag cttggagaga ctctcgaccc gcttctccga
     5221 ttcgccgcgg gcaacgaagt cgcgcaggcg ccggatgatc tggccggcgc gcagcgcctg
     5281 ctcggcggcg cggtccaggg cgctttcgac cttcggtgtg ttcggatcac tgctgccggc
     5341 aagcagccgc cgcgagccct tcatgtagtt gctgatcgcc gccagcggct ggttgagctc
     5401 gtgcgcgagc gcggacgcca tttcgcccat ggcgctcagc ctggagacgt ggacgagctc
     5461 ggattgcagt tcctggaggc gcgcctgggt ctgctggtgc tcggtgatat cattctgaat
     5521 tccgacaaaa tacgttttat cctctatttc cattggatca atatttaatt cattccagaa
     5581 catcgttccg tcttttttgt agttttggat ctgaacggtg accggttctt tattttgtaa
     5641 agcggttctg atgttgtcca cttctgcagg atctgtgtgt ttcccctgta agaagcgaca
     5701 gttctttcct aaaatttcct cggtctcgta gccggtcatt tgaacaaagc cttgatttac
     5761 gtagacaata ggattatctt caagtgcggg atctgtaatt accacaccga ctcgcacgtg
     5821 atcaagtgct tttttgatga cttccagctg tcctggtatc ccaaatgatt gaaaactagc
     5881 cacattcacc accctcaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt
     5941 tttgcaccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta
     6001 ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat
     6061 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     6121 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     6181 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     6241 agaggatcga gatctacgcc cgtgatcctg atcaccggct atccggacga aaacatctcg
     6301 acccgggccg ccgaggccgg cgtaaaagac gtggttttga agccgcttct cgacgaaaac
     6361 ctgctcaagc gtatccgccg cgccatccag gaccggcctc gggcatgacc tacggggttc
     6421 tacgtaaggc acccccctta agatatcgct cgaaattttc gaacctcccg ataccgcgta
     6481 ccaatgcgtc atcacaacgg agtactagta aagaggagaa atactagatg agcacaaaaa
     6541 agaaaccatt aacacaagag cagcttgagg acgcacgtcg ccttaaagca atttatgaaa
     6601 aaaagaaaaa tgaacttggc ttatcccagg aatctgtcgc agacaagatg gggatggggc
     6661 agtcaggcgt tggtgcttta tttaatggca tcaatgcatt aaatgcttat aacgccgcat
     6721 tgcttgcaaa aattctcaaa gttagcgttg aagaatttag cccttcaatc gccagagaaa
     6781 tctacgagat gtatgaagcg gttagtatgc agccgtcact tagaagtgag tatgagtacc
     6841 ctgttttttc tcatgttcag gcagggatgt tctcacctga gcttagaacc tttaccaaag
     6901 gtgatgcgga gagatgggta agcacaacca aaaaagccag tgattctgca ttctggcttg
     6961 aggttgaagg taattccatg accgcaccaa caggctccaa gccgagcttt cctgacggaa
     7021 tgttaattct cgttgaccct gagcaggctg ttgagccagg tgatttctgc atagccagac
     7081 ttgggggtga tgagtttacc ttcaagaaac tgatcaggga tagcggtcag gtgtttttac
     7141 aaccactaaa cccacagtac ccaatgatcc catgcaatga gagttgttcc gttgtgggga
     7201 aagttatcgc tagtcagtgg cctgaagaga cgtttggcgc tgcaaacgac gaaaactacg
     7261 ctttagtagc ttaataacgc tgatagtgct agtgtagatc gctactagag ccaggcatca
     7321 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     7381 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttatat
     7441 actagagtaa caccgtgcgt gttgactatt ttacctctgg cggtgataat ggttgctcta
     7501 gaaataattt tgtttaactt taagaaggag agctagcatg aaaccagtaa cgttatacga
     7561 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     7621 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     7681 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     7741 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     7801 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     7861 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     7921 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     7981 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     8041 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     8101 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     8161 tcaaattcag ccgatagcgg aacgggaagg
//
