LOCUS       sequence-422483-        4140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Unit Bactoneuron-9, expresses EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4140)
  AUTHORS   Sarkar K, Bonnerjee D, Srivastava R, Bagh S
  TITLE     A single layer artificial neural network type architecture with 
            molecular engineered bacteria for reversible and irreversible 
            computing.
  JOURNAL   Chem Sci. 2021 Nov 9;12(48):15821-15832. doi: 10.1039/d1sc01505b. 
            eCollection 2021 Dec 15.
  PUBMED    35024106
REFERENCE   2  (bases 1 to 4140)
  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: 
            "10.1039/d1sc01505b"; journalName: "Chem Sci"; date: "2021-11-9- 9";
            volume: "12"; issue: "48"; pages: "15821-15832"
COMMENT     Sequence Label: Bneu-9
FEATURES             Location/Qualifiers
     source          1..4140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(78..869,870..938))
                     /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     701..720
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(939..1043)
                     /gene="bla"
                     /label=AmpR promoter
     terminator      complement(1155..1241)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(1265..1978,1979..1981,1982..1984))
                     /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             complement(join(1268..1978,1979..1981,1982..1984))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1310..1331)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1659..1678
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1918..1939
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     RBS             1991..2002
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    2012..2030
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2042..2060
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2148..2166
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2187..2221
                     /label=BBa_J23102
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.86"
     RBS             2231..2242
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      3059..3145
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3418..4006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3498..3517)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(join(4094..4140,1..48))
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctcg
       61 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
      121 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
      181 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
      241 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
      301 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
      361 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
      421 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
      481 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
      541 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
      601 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
      661 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
      721 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
      781 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
      841 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
      901 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
      961 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     1021 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctactgg
     1081 gacgaagacg aacagtcgac aacgaagacg ccaacatcaa gatctcgcat ttgttcctag
     1141 gtctagggcg gcggatttgt cctactcagg agagcgttca ccgacaaaca acagataaaa
     1201 cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctcta gagtcgcggc
     1261 cgctttactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca
     1321 gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca
     1381 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
     1441 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga
     1501 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
     1561 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
     1621 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
     1681 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
     1741 gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca
     1801 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
     1861 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt
     1921 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
     1981 ccatggtacc tttctcctct ttaatgaatt ctctctatca ctgataggga tgtgctcagt
     2041 atctctatca ctgataggga tgtcaatatc accgccagag gtaatctatc accgccagag
     2101 gtactcgagt taccaggatc tttattatca tgacattaac ctataaaaat aggcgtatca
     2161 cgaggccctt tcgtcttcac ctcgagttga cagctagctc agtcctaggt actgtgctag
     2221 cgaattcatt aaagaggaga aaggtaccat gaaaaacata aatgccgacg acacatacag
     2281 aataattaat aaaattaaag cttgtagaag caataatgat attaatcaat gcttatctga
     2341 tatgactaaa atggtacatt gtgaatatta tttactcgcg atcatttatc ctcattctat
     2401 ggttaaatct gatatttcaa tcctagataa ttaccctaaa aaatggaggc aatattatga
     2461 tgacgctaat ttaataaaat atgatcctat agtagattat tctaactcca atcattcacc
     2521 aattaattgg aatatatttg aaaacaatgc tgtaaataaa aaatctccaa atgtaattaa
     2581 agaagcgaaa acatcaggtc ttatcactgg gtttagtttc cctattcata cggctaacaa
     2641 tggcttcgga atgcttagtt ttgcacattc agaaaaagac aactatatag atagtttatt
     2701 tttacatgcg tgtatgaaca taccattaat tgttccttct ctagttgata attatcgaaa
     2761 aataaatata gcaaataata aatcaaacaa cgatttaacc aaaagagaaa aagaatgttt
     2821 agcgtgggca tgcgaaggaa aaagctcttg ggatatttca aaaatattag gttgcagtga
     2881 gcgtactgtc actttccatt taaccaatgc gcaaatgaaa ctcaatacaa caaaccgctg
     2941 ccaaagtatt tctaaagcaa ttttaacagg agcaattgat tgcccatact ttaaaaatta
     3001 ataacactga tagtgctagt gtagatcact actagagcca ggcatcatct agaggcatca
     3061 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     3121 gaacgctctc ctgagtagga caaatccgcc gccctagacc taggcgatga cggaaaaaga
     3181 gatcgtctta atgtatagat tttcgtggat tacgtcgcca gtcaggatcc tggcagaagc
     3241 tattcattct gcagttccgt gctccaaaac aaccgttcgg ctgcggcgag cggtatcagc
     3301 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3361 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     3421 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     3481 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     3541 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     3601 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3661 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3721 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3781 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3841 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3901 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3961 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4021 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4081 gactagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca
//
