LOCUS       sequence-500608-        3116 bp    DNA     circular SYN 20-FEB-2026
DEFINITION  EGFP expressed under PL LacO1 promoter; part of BNeu AS11; colE1 
            origin of replication.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3116)
  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 3116)
  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_PLLacO1_R1_EGFP
FEATURES             Location/Qualifiers
     source          1..3116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    9..25
                     /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    32..48
                     /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             90..101
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(108..110,111..113,114..827)
                     /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(108..110,111..113,114..824)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(153..174)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(414..433)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     761..782
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      851..937
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(1210..1798)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1290..1309)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1886..1980)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2010..2801,2802..2870))
                     /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     2633..2652
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2871..2975)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3084..3102
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaatt gtgagcggat aacaattgac attgtgagcg gataacaaga tactgagcac
       61 atcagcagga cgcactgacc gaattcatta aagaggagaa aggtaccatg gtgagcaagg
      121 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
      181 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
      241 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
      301 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
      361 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
      421 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
      481 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
      541 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
      601 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
      661 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
      721 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
      781 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact
      841 ctagaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
      901 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgccctaga cctaggcgat
      961 gacggaaaaa gagatcgtct taatgtatag attttcgtgg attacgtcgc cagtcaggat
     1021 cctggcagaa gctattcatt ctgcagttcc gtgctccaaa acaaccgttc ggctgcggcg
     1081 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1141 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1201 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1261 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1321 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1381 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1441 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1501 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1561 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1621 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     1681 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1741 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1801 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1861 gattttggtc atgactagtg cttggattct caccaataaa aaacgcccgg cggcaaccga
     1921 gcgttctgaa caaatccaga tggagttctg aggtcattac tggatctatc aacaggagtc
     1981 caagcgagct cgtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2041 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2101 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2161 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2221 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2281 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2341 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2401 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2461 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2521 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2581 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2641 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2701 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2761 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2821 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2881 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2941 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3001 gacgtctact gggacgaaga cgaacagtcg acaacgaaga cgccaacatc aagatcttta
     3061 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcac
//
