LOCUS       sequence-343723-        8810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes OptoLAC2B 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: pMAL659
FEATURES             Location/Qualifiers
     source          1..8810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..19)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      172..258
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      350..377
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        396..487
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      945..1400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1032..1051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1242..1263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1956..2615)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2530..2549
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2616..2718)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3110..3127)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3244..3789)
                     /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(3324..3343)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4026..4073)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4069..4087
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4140..4157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4198..5277)
                     /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     5239..5258
                     /label=LacI-R
                     /note="LacI, reverse primer"
     RBS             5284..5306
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      complement(5382..5409)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(5425..5496)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5544..5576)
                     /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(5577..6287)
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="MSTKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQ
                     SGVGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEY
                     PVFSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPD
                     GMLILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSV
                     VGKVIASQWPEETFG"
     RBS             6294..6305
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        6854..6931
                     /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 gcctgataca gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg
       61 gcagtagcgc ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg
      121 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
      181 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
      241 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
      301 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
      361 ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct aaatacattc
      421 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      481 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      541 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      601 gggtgcagca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
      661 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
      721 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
      781 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
      841 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
      901 taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg
      961 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1021 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1081 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1141 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1201 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1261 tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     1321 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     1381 aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1441 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1501 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1561 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1621 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     1681 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1741 cagtcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     1801 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa
     1861 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaacca
     1921 ggcgtttaag ggcaccaata actgccttaa aaaaattacg ccccgccctg ccactcatcg
     1981 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
     2041 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
     2101 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
     2161 ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt agggaaatag
     2221 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
     2281 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
     2341 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacggaat
     2401 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
     2461 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
     2521 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     2581 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     2641 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     2701 cctcttacgt gccgatcaac gtctcatttt cgccaaaagt tggcccaggg cttcccggta
     2761 tcaacaggga caccaggatt tatttattct gcgaagtgat cttccgtcac aggtatttat
     2821 tcggcgcaaa gtgcgtcggg tgatgctgcc aacttactga tttagtgtat gatggtgttt
     2881 ttgaggtgct ccagtggctt ctgtttctat cagctgtccc tcctgttcag ctactgacgg
     2941 ggtggtgcgt aacggcaaaa gcaccgccgg acatcagcgc tagcggagtg tatactggct
     3001 tactatgttg gcactgatga gggtgtcagt gaagtgcttc atgtggcagg agaaaaaagg
     3061 ctgcaccggt gcgtcagcag aatatgtgat acaggatata ttccgcttcc tcgctcactg
     3121 actcgctacg ctcggtcgtt cgactgcggc gagcggaaat ggcttacgaa cggggcggag
     3181 atttcctgga agatgccagg aagatactta acagggaagt gagagggccg cggcaaagcc
     3241 gtttttccat aggctccgcc cccctgacaa gcatcacgaa atctgacgct caaatcagtg
     3301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggcg gctccctcgt
     3361 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt
     3421 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta
     3481 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3541 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag
     3601 gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac
     3661 tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa
     3721 aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa
     3781 cgatctcaag aagatcatct tattaatcag ataaaatatt tctaggctca tgagcccgaa
     3841 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     3901 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatctgct catgtttgac
     3961 agcttatcat cgatgcatgc ggccgccgtc ccattcgcca atccggatat agttcctcct
     4021 ttcagcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc
     4081 tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt tgttagcagc cggatctcag
     4141 tggtggtggt ggtggtgctc attaaactgc tgcagcgtag ttttcgtcgt ttgctgcctg
     4201 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4261 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     4321 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     4381 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     4441 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     4501 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     4561 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     4621 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     4681 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     4741 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     4801 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     4861 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     4921 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     4981 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     5041 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     5101 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     5161 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     5221 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcatgct
     5281 agctctcctt cttaaagtta aacaaaatta tttctagagc aaccattatc accgccagag
     5341 gtaaaatagt caacacgcac ggtgttactc tagtatataa acgcagaaag gcccacccga
     5401 aggtgagcca gtgtgactct agtagagagc gttcaccgac aaacaacaga taaaacgaaa
     5461 ggcccagtct ttcgactgag cctttcgttt tatttgatgc ctggctctag tagcgatcta
     5521 cactagcact atcagcgtta ttaagctact aaagcgtagt tttcgtcgtt tgcagcgcca
     5581 aacgtctctt caggccactg actagcgata actttcccca caacggaaca actctcattg
     5641 catgggatca ttgggtactg tgggtttagt ggttgtaaaa acacctgacc gctatccctg
     5701 atcagtttct tgaaggtaaa ctcatcaccc ccaagtctgg ctatgcagaa atcacctggc
     5761 tcaacagcct gctcagggtc aacgagaatt aacattccgt caggaaagct cggcttggag
     5821 cctgttggtg cggtcatgga attaccttca acctcaagcc agaatgcaga atcactggct
     5881 tttttggttg tgcttaccca tctctccgca tcacctttgg taaaggttct aagctcaggt
     5941 gagaacatcc ctgcctgaac atgagaaaaa acagggtact catactcact tctaagtgac
     6001 ggctgcatac taaccgcttc atacatctcg tagatttctc tggcgattga agggctaaat
     6061 tcttcaacgc taactttgag aatttttgca agcaatgcgg cgttataagc atttaatgca
     6121 ttgatgccat taaataaagc accaacgcct gactgcccca tccccatctt gtctgcgaca
     6181 gattcctggg ataagccaag ttcatttttc tttttttcat aaattgcttt aaggcgacgt
     6241 gcgtcctcaa gctgctcttg tgttaatggt ttcttttttg tgctcatcta gtatttctcc
     6301 tctttactag tactccgttg tgatgacgca ttggtacgcg gtatcgggag gttcgaaaat
     6361 ttcgagcgat atcttaaggg gggtgcctta cgtagaaccc cgtaggtcat gcccgaggcc
     6421 ggtcctggat ggcgcggcgg atacgcttga gcaggttttc gtcgagaagc ggcttcaaaa
     6481 ccacgtcttt tacgccggcc tcggcggccc gggtcgagat gttttcgtcc ggatagccgg
     6541 tgatcaggat cacgggcgta gatctcgatc ctctacgccg gacgcatcgt ggccggcatc
     6601 accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat
     6661 cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc
     6721 gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg
     6781 ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag
     6841 cgtcgagatc ccggacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg
     6901 cccggaagag agtcaattga gggtggtgaa tgtggctagt tttcaatcat ttgggatacc
     6961 aggacagctg gaagtcatca aaaaagcact tgatcacgtg cgagtcggtg tggtaattac
     7021 agatcccgca cttgaagata atcctattgt ctacgtaaat caaggctttg ttcaaatgac
     7081 cggctacgag accgaggaaa ttttaggaaa gaactgtcgc ttcttacagg ggaaacacac
     7141 agatcctgca gaagtggaca acatcagaac cgctttacaa aataaagaac cggtcaccgt
     7201 tcagatccaa aactacaaaa aagacggaac gatgttctgg aatgaattaa atattgatcc
     7261 aatggaaata gaggataaaa cgtattttgt cggaattcag aatgatatca ccgagcacca
     7321 gcagacccag gcgcgcctcc aggaactgca atccgagctc gtccacgtct ccaggctgag
     7381 cgccatgggc gaaatggcgt ccgcgctcgc gcacgagctc aaccagccgc tggcggcgat
     7441 cagcaactac atgaagggct cgcggcggct gcttgccggc agcagtgatc cgaacacacc
     7501 gaaggtcgaa agcgccctgg accgcgccgc cgagcaggcg ctgcgcgccg gccagatcat
     7561 ccggcgcctg cgcgacttcg ttgcccgcgg cgaatcggag aagcgggtcg agagtctctc
     7621 caagctgatc gaggaggccg gcgcgctcgg gcttgccggc gcgcgcgagc agaacgtgca
     7681 gctccgcttc agtctcgatc cgggcgccga tctcgttctc gccgaccggg tgcagatcca
     7741 gcaggtcctg gtcaacctgt tccgcaacgc gctggaagcg atggctcagt cgcagcgacg
     7801 cgagctcgtc gtcaccaaca cccccgccgc cgacgacatg atcgaggtcg aagtgtccga
     7861 caccggcagc ggtttccagg acgacgtcat tccgaacctg tttcagactt tcttcaccac
     7921 caaggacacc ggcatgggcg tgggactgtc catcagccgc tcgatcatcg aagctcacgg
     7981 cgggcgcatg tgggccgaga gcaacgcatc gggcggggcg accttccgct tcaccctccc
     8041 ggcagccgac gagatgatag gaggtctagc atgacgacca agggacatat ctacgtcatc
     8101 gacgacgacg cggcgatgcg ggattcgctg aatttcctgc tggattctgc cggcttcggc
     8161 gtcacgctgt ttgacgacgc gcaagccttt ctcgacgccc tgccgggtct ctccttcggc
     8221 tgcgtcgtct ccgacgtgcg catgccgggc cttgacggca tcgagctgtt gaagcggatg
     8281 aaggcgcagc aaagcccctt tccgatcctc atcatgaccg gtcacggcga cgtgccgctc
     8341 gcggtcgagg cgatgaagtt aggggcggtg gactttctgg aaaagccttt cgaggacgac
     8401 cgcctcaccg ccatgatcga atcggcgatc cgccaggccg agccggccgc caagagcgag
     8461 gccgtcgcgc aggatatcgc cgcccgcgtc gcctcgttga gccccaggga gcgccaggtc
     8521 atggaagggc tgatcgccgg cctttccaac aagctgatcg cccgcgagta cgacatcagc
     8581 ccgcgcacca tcgaggtgta tcgggccaac gtcatgacca agatgcaggc caacagcctt
     8641 tcggagctgg ttcgcctcgc gatgcgcgcc ggcatgctca acgattgaca attgatgtaa
     8701 gttagctcac tcattaggca ccgggatctc gaccgatgcc cttgagagcc ttcaacccag
     8761 tctgcaggca tgcaagcttg gctgttttgg cggatgagag aagattttca
//
