LOCUS       sequence-502191-        4046 bp    DNA     circular SYN 09-MAR-2026
DEFINITION  Cassette 1: Mutated cI protein expressed under PL TetO1 promoter; 
            Cassette 2: Mutated cI protein expressed under PL LacO1 promoter; 
            part of BNeu AS10; colE1 origin of replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4046)
  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 4046)
  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: pM2_PLLaco1R1CI*_PLTetO1R1CI*
FEATURES             Location/Qualifiers
     source          1..4046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(7..25,26..31,32..48,49..54,55..80)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    7..25
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    32..50
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             90..101
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             113..820
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="TKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQSG
                     VGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEYPV
                     FSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPDGM
                     LILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSVVG
                     KVIASQWPEETFG"
     terminator      832..918
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(1033..1119)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1131..1838)
                     /codon_start=1
                     /gene="cI"
                     /product="phage lambda repressor"
                     /label=lambda repressor
                     /translation="TKKKPLTQEQLEDARRLKAIYEKKKNELGLSQESVADKMGMGQSG
                     VGALFNGINALNAYNAALLAKILKVSVEEFSPSIAREIYEMYEAVSMQPSLRSEYEYPV
                     FSHVQAGMFSPELRTFTKGDAERWVSTTKKASDSAFWLEVEGNSMTAPTGSKPSFPDGM
                     LILVDPEQAVEPGDFCIARLGGDEFTFKKLIRDSGQVFLQPLNPQYPMIPCNESCSVVG
                     KVIASQWPEETFG"
     RBS             1850..1861
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    1903..1919
                     /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    1926..1942
                     /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)."
     primer_bind     complement(1965..1983)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2092..2196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2197..2265,2266..3057)
                     /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(2415..2434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3087..3181
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3269..3857
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3758..3777
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcgagtccc tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac
       61 atcagcagga cgcactgacc gaattcatta aagaggagaa aggtaccatg gcacaaaaaa
      121 gaaaccatta acacaagagc agcttgagga cgcacgtcgc cttaaagcaa tttatgaaaa
      181 aaagaaaaat gaacttggct tatcccagga atctgtcgca gacaagatgg ggatggggca
      241 gtcaggcgtt ggtgctttat ttaatggcat caatgcatta aatgcttata acgccgcatt
      301 gcttgcaaaa attctcaaag ttagcgttga agaatttagc ccttcaatcg ccagagaaat
      361 ctacgagatg tatgaagcgg ttagtatgca gccgtcactt agaagtgagt atgagtaccc
      421 tgttttttct catgttcagg cagggatgtt ctcacctgag cttagaacct ttaccaaagg
      481 tgatgcggag agatgggtaa gcacaaccaa aaaagccagt gattctgcat tctggcttga
      541 ggttgaaggt aattccatga ccgcaccaac aggctccaag ccaagctttc ctgacggaat
      601 gttaattctc gttgaccctg agcaggctgt tgagccaggt gatttctgca tagccagact
      661 tgggggtgat gagtttacct tcaagaaact gatcagggat agcggtcagg tgtttttaca
      721 accactaaac ccacagtacc caatgatccc atgcaatgag agttgttccg ttgtggggaa
      781 agttatcgct agtcagtggc ctgaagagac gtttggctga tctagaggca tcaaataaaa
      841 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
      901 ctcctgagta ggacaaatcc gccgccctag acctaggaac aaatgcgaga tcctgactgg
      961 cgacgtaatc cacgaaaatc tatacattaa gacgatctct ttttccgtca tcgcctaggt
     1021 ctagggcggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     1081 aaaggcccag tctttcgact gagcctttcg ttttatttga tgcctctaga tcagccaaac
     1141 gtctcttcag gccactgact agcgataact ttccccacaa cggaacaact ctcattgcat
     1201 gggatcattg ggtactgtgg gtttagtggt tgtaaaaaca cctgaccgct atccctgatc
     1261 agtttcttga aggtaaactc atcaccccca agtctggcta tgcagaaatc acctggctca
     1321 acagcctgct cagggtcaac gagaattaac attccgtcag gaaagcttgg cttggagcct
     1381 gttggtgcgg tcatggaatt accttcaacc tcaagccaga atgcagaatc actggctttt
     1441 ttggttgtgc ttacccatct ctccgcatca cctttggtaa aggttctaag ctcaggtgag
     1501 aacatccctg cctgaacatg agaaaaaaca gggtactcat actcacttct aagtgacggc
     1561 tgcatactaa ccgcttcata catctcgtag atttctctgg cgattgaagg gctaaattct
     1621 tcaacgctaa ctttgagaat ttttgcaagc aatgcggcgt tataagcatt taatgcattg
     1681 atgccattaa ataaagcacc aacgcctgac tgccccatcc ccatcttgtc tgcgacagat
     1741 tcctgggata agccaagttc atttttcttt ttttcataaa ttgctttaag gcgacgtgcg
     1801 tcctcaagct gctcttgtgt taatggtttc ttttttgtgc catggtacct ttctcctctt
     1861 taatgaattc ggtcagtgcg tcctgctgat gtgctcagta tcttgttatc cgctcacaat
     1921 gtcaattgtt atccgctcac aattctcgag gtgaagacga aagggcctcg tgatacgcct
     1981 atttttatag gttaatgtca tgataataaa gatcttgatg ttggcgtctt cgttgtcgac
     2041 tgttcgtctt cgtcccagta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2101 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2161 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     2221 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     2281 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2341 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2401 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     2461 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2521 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2581 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2641 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2701 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2761 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2821 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2881 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2941 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3001 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3061 tcagaccaag tttacgagct cgcttggact cctgttgata gatccagtaa tgacctcaga
     3121 actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa
     3181 tccaagcact agtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     3241 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3301 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3361 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     3421 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3481 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3541 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3601 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3661 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3721 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3781 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     3841 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     3901 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     3961 ccgcctttga gtgagctgat accgctcgcc gcagccgaac ggttgttttg gagcacggaa
     4021 ctgcagttct accaggatcc tggtaa
//
