LOCUS       sequence-502190-        7089 bp    DNA     circular SYN 09-MAR-2026
DEFINITION  LuxR expressed under BBa_J23102; E2-Crimson under PLC promoter; 
            mutated cI* under PLTetO1; and cI* under Plux*; part of BNeuAS5; pUC
            origin of replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7089)
  AUTHORS   Bonnerjee D, Chakraborty S, Mukherjee B, Basu R, Paul A, Bagh S
  TITLE     Multicellular artificial neural network-type architectures 
            demonstrate computational problem solving.
  JOURNAL   Nat Chem Biol. 2024 Nov;20(11):1524-1534. doi: 
            10.1038/s41589-024-01711-4. Epub 2024 Sep 16.
  PUBMED    39285005
REFERENCE   2  (bases 1 to 7089)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 9, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41589-024-01711-4"; journalName: "Nat Chem Biol"; date: 
            "2024-11"; volume: "20"; issue: "11"; pages: "1524-1534"
COMMENT     Sequence Label: pM3_PLux*R1CI*_PLTetO1R1CI*_PLCR1E2Cr_PJ23102R...
FEATURES             Location/Qualifiers
     source          1..7089
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1..87)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             904..915
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(925..959)
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     protein_bind    1045..1061
                     /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)."
     protein_bind    1068..1084
                     /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             1172..1183
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1190..1867
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     1588..1607
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     terminator      complement(1882..1928)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1940..2647)
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="TKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQSG
                     VGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEYPV
                     FSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPDGM
                     LILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSVVG
                     KVIASQWPEETFG"
     RBS             2659..2670
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(join(2680..2705,2706..2711,2712..2728,2729..2734,
                     2735..2753))
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    2710..2728
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2735..2753
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2901..2912
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2924..3631
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="TKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQSG
                     VGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEYPV
                     FSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPDGM
                     LILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSVVG
                     KVIASQWPEETFG"
     terminator      3643..3729
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        3841..3945
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3946..4014,4015..4806)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4164..4183)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      4836..4930
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      5248..5836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      6229..6817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
       61 tttcgactga gcctttcgtt ttatttgatg cctctagatg atgcctggct ctagtagtga
      121 tctacactag cactatcagt gttattaatt tttaaagtat gggcaatcaa ttgctcctgt
      181 taaaattgct ttagaaatac tttggcagcg gtttgttgta ttgagtttca tttgcgcatt
      241 ggttaaatgg aaagtgacag tacgctcact gcaacctaat atttttgaaa tatcccaaga
      301 gctttttcct tcgcatgccc acgctaaaca ttctttttct cttttggtta aatcgttgtt
      361 tgatttatta tttgctatat ttatttttcg ataattatca actagagaag gaacaattaa
      421 tggtatgttc atacacgcat gtaaaaataa actatctata tagttgtctt tttctgaatg
      481 tgcaaaacta agcattccga agccattgtt agccgtatga atagggaaac taaacccagt
      541 gataagacct gatgttttcg cttctttaat tacatttgga gattttttat ttacagcatt
      601 gttttcaaat atattccaat taattggtga atgattggag ttagaataat ctactatagg
      661 atcatatttt attaaattag cgtcatcata atattgcctc cattttttag ggtaattatc
      721 taggattgaa atatcagatt taaccataga atgaggataa atgatcgcga gtaaataata
      781 ttcacaatgt accattttag tcatatcaga taagcattga ttaatatcat tattgcttct
      841 acaagcttta attttattaa ttattctgta tgtgtcgtcg gcatttatgt ttttcatggt
      901 acctttctcc tctttaatga attcgctagc acagtaccta ggactgagct agctgtcaac
      961 tcgaggtgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     1021 ataaagatcc tggtaactcg agaattgtga gcggataaca attgacattg tgagcggata
     1081 acaagatact gagcacatct acctctggcg gtgatagatg attacctctg gcggtgatag
     1141 atgattacct ctggcggtga tagaattcat taaagaggag aaaggtacca tggatagcac
     1201 tgagaacgtc atcaagccct tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
     1261 ccacgagttc gagatcgagg gcgtgggcga gggcaagccc tacgagggca cccagaccgc
     1321 caagctgcaa gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     1381 gttcttctac ggctccaagg cgtacatcaa gcaccccgcc gacatccccg actacctcaa
     1441 gcagtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     1501 ggtgaccgtg acccaggact cctccctgca ggacggcacc ctcatctacc acgtgaagtt
     1561 catcggcgtg aacttcccct ccgacggccc cgtaatgcag aagaagactc tgggctggga
     1621 gccctccact gagcgcaact acccccgcga cggcgtgctg aagggcgaga accacatggc
     1681 gctgaagctg aagggcggcg gccactacct gtgtgagttc aagtccatct acatggccaa
     1741 gaagcccgtg aagctgcccg gctaccacta cgtggactac aagctcgaca tcacctccca
     1801 caacgaggac tacaccgtgg tggagcagta cgagcgcgcc gaggcccgcc accacctgtt
     1861 ccagtagtct agaggcatca aataaaacga aaggcccagt ctttcgactg agcctttcgt
     1921 tttatttgat gcctctagat cagccaaacg tctcttcagg ccactgacta gcgataactt
     1981 tccccacaac ggaacaactc tcattgcatg ggatcattgg gtactgtggg tttagtggtt
     2041 gtaaaaacac ctgaccgcta tccctgatca gtttcttgaa ggtaaactca tcacccccaa
     2101 gtctggctat gcagaaatca cctggctcaa cagcctgctc agggtcaacg agaattaaca
     2161 ttccgtcagg aaagcttggc ttggagcctg ttggtgcggt catggaatta ccttcaacct
     2221 caagccagaa tgcagaatca ctggcttttt tggttgtgct tacccatctc tccgcatcac
     2281 ctttggtaaa ggttctaagc tcaggtgaga acatccctgc ctgaacatga gaaaaaacag
     2341 ggtactcata ctcacttcta agtgacggct gcatactaac cgcttcatac atctcgtaga
     2401 tttctctggc gattgaaggg ctaaattctt caacgctaac tttgagaatt tttgcaagca
     2461 atgcggcgtt ataagcattt aatgcattga tgccattaaa taaagcacca acgcctgact
     2521 gccccatccc catcttgtct gcgacagatt cctgggataa gccaagttca tttttctttt
     2581 tttcataaat tgctttaagg cgacgtgcgt cctcaagctg ctcttgtgtt aatggtttct
     2641 tttttgtgcc atggtacctt tctcctcttt aatgaattcg gtcagtgcgt cctgctgatg
     2701 tgctcagtat ctctatcact gatagggatg tcaatctcta tcactgatag ggactcgagg
     2761 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataaag
     2821 atcctggtaa ctcgagacct gtaggatcgt acaggtttac gcaagaaaat ggtttgttac
     2881 tttcgaataa agaattcatt aaagaggaga aaggtaccat ggcacaaaaa agaaaccatt
     2941 aacacaagag cagcttgagg acgcacgtcg ccttaaagca atttatgaaa aaaagaaaaa
     3001 tgaacttggc ttatcccagg aatctgtcgc agacaagatg gggatggggc agtcaggcgt
     3061 tggtgcttta tttaatggca tcaatgcatt aaatgcttat aacgccgcat tgcttgcaaa
     3121 aattctcaaa gttagcgttg aagaatttag cccttcaatc gccagagaaa tctacgagat
     3181 gtatgaagcg gttagtatgc agccgtcact tagaagtgag tatgagtacc ctgttttttc
     3241 tcatgttcag gcagggatgt tctcacctga gcttagaacc tttaccaaag gtgatgcgga
     3301 gagatgggta agcacaacca aaaaagccag tgattctgca ttctggcttg aggttgaagg
     3361 taattccatg accgcaccaa caggctccaa gccaagcttt cctgacggaa tgttaattct
     3421 cgttgaccct gagcaggctg ttgagccagg tgatttctgc atagccagac ttgggggtga
     3481 tgagtttacc ttcaagaaac tgatcaggga tagcggtcag gtgtttttac aaccactaaa
     3541 cccacagtac ccaatgatcc catgcaatga gagttgttcc gttgtgggga aagttatcgc
     3601 tagtcagtgg cctgaagaga cgtttggctg atctagaggc atcaaataaa acgaaaggct
     3661 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     3721 aggacaaatc cgccgcccta gacctaggaa caaatgcgag atcttgatgt tggcgtcttc
     3781 gttgtcgact gttcgtcttc gtcccagtag acgtcaggtg gcacttttcg gggaaatgtg
     3841 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     3901 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     3961 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4021 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4081 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4141 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4201 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4261 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     4321 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     4381 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     4441 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     4501 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     4561 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     4621 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     4681 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     4741 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     4801 tggtaactgt cagaccaagt ttacgagctc gcttggactc ctgttgatag atccagtaat
     4861 gacctcagaa ctccatctgg atttgttcag aacgctcggt tgccgccggg cgttttttat
     4921 tggtgagaat ccaagcacta gtggctggca ctctgtcgat accccaccga gaccccattg
     4981 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     5041 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     5101 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5161 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5221 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5281 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5341 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5401 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5461 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     5521 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     5581 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5641 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5701 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5761 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5821 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5881 gccttttgct cacatgttct ttcctgcgtt atcccctact agtggctggc actctgtcga
     5941 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     6001 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     6061 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6121 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6181 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6241 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6301 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6361 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6421 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6481 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6541 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6601 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6661 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6721 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6781 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6841 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6901 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6961 ggttgttttg gagcacggaa ctgcagaatg aatagcttct gccaggatcc tgactggcga
     7021 cgtaatccac gaaaatctat acattaagac gatctctttt tccgtcatcg cctaggtcta
     7081 gggcggcgg
//
