LOCUS       sequence-500599-        4140 bp    DNA     circular SYN 20-FEB-2026
DEFINITION  EGFP expressed under PCT synthetic promoter and LuxR expressed under
            BBa_J23102 promoter; component of BNeuAS3; colE1 origin of 
            replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4140)
  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 4140)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 20, 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_PCTR1EGFP_PJ23102R1LuxR
FEATURES             Location/Qualifiers
     source          1..4140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             760..771
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(781..815)
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     primer_bind     complement(836..854)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    942..960
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    972..990
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1000..1011
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1018..1020,1021..1023,1024..1737)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1018..1020,1021..1023,1024..1734)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1063..1084)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1324..1343)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1671..1692
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1761..1847
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        1959..2063
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2064..2132,2133..2924)
                     /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(2282..2301)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      2954..3048
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3136..3724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3625..3644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3997..4083)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattttta aagtatgggc aatcaattgc tcctgttaaa attgctttag aaatactttg
       61 gcagcggttt gttgtattga gtttcatttg cgcattggtt aaatggaaag tgacagtacg
      121 ctcactgcaa cctaatattt ttgaaatatc ccaagagctt tttccttcgc atgcccacgc
      181 taaacattct ttttctcttt tggttaaatc gttgtttgat ttattatttg ctatatttat
      241 ttttcgataa ttatcaacta gagaaggaac aattaatggt atgttcatac acgcatgtaa
      301 aaataaacta tctatatagt tgtctttttc tgaatgtgca aaactaagca ttccgaagcc
      361 attgttagcc gtatgaatag ggaaactaaa cccagtgata agacctgatg ttttcgcttc
      421 tttaattaca tttggagatt ttttatttac agcattgttt tcaaatatat tccaattaat
      481 tggtgaatga ttggagttag aataatctac tataggatca tattttatta aattagcgtc
      541 atcataatat tgcctccatt ttttagggta attatctagg attgaaatat cagatttaac
      601 catagaatga ggataaatga tcgcgagtaa ataatattca caatgtacca ttttagtcat
      661 atcagataag cattgattaa tatcattatt gcttctacaa gctttaattt tattaattat
      721 tctgtatgtg tcgtcggcat ttatgttttt catggtacct ttctcctctt taatgaattc
      781 gctagcacag tacctaggac tgagctagct gtcaactcga ggtgaagacg aaagggcctc
      841 gtgatacgcc tatttttata ggttaatgtc atgataataa agatcctggt aactcgagta
      901 cctctggcgg tgatagatta cctctggcgg tgatattgac atccctatca gtgatagaga
      961 tactgagcac atccctatca gtgatagaga gaattcatta aagaggagaa aggtaccatg
     1021 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     1081 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     1141 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     1201 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag
     1261 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     1321 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     1381 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     1441 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc
     1501 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac
     1561 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     1621 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     1681 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc
     1741 ggccgcgact ctagaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     1801 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgccctaga
     1861 cctaggaaca aatgcgagat cttgatgttg gcgtcttcgt tgtcgactgt tcgtcttcgt
     1921 cccagtagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     1981 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2041 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2101 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2161 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2221 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2281 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2341 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2401 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2461 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2521 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2581 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2641 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2701 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2761 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2821 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2881 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2941 acgagctcgc ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat
     3001 ttgttcagaa cgctcggttg ccgccgggcg ttttttattg gtgagaatcc aagcactagt
     3061 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     3121 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3181 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3241 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     3301 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3361 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3421 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3481 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3541 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     3601 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     3661 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     3721 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     3781 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     3841 agctgatacc gctcgccgca gccgaacggt tgttttggag cacggaactg cagaatgaat
     3901 agcttctgcc aggatcctga ctggcgacgt aatccacgaa aatctataca ttaagacgat
     3961 ctctttttcc gtcatcgcct aggtctaggg cggcggattt gtcctactca ggagagcgtt
     4021 caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct ttcgttttat
     4081 ttgatgcctc tagatgatgc ctggctctag tagtgatcta cactagcact atcagtgtta
//
