LOCUS       sequence-419677-        4398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Unit Bactoneuron-10, expresses E2-crimson.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4398)
  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 4398)
  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-10
FEATURES             Location/Qualifiers
     source          1..4398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(273..359)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             1176..1187
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(1197..1231)
                     /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(1252..1270)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    1317..1333
                     /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    1340..1356
                     /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             1444..1455
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1462..1464,1465..1467,1468..2181)
                     /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(1462..1464,1465..1467,1468..2178)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1507..1528)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1768..1787)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2115..2136
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2205..2291
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        2403..2507
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2508..2576,2577..3368)
                     /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(2726..2745)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     terminator      3398..3492
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3810..4398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4299..4318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacggttgtt ttggagcacg gaactgcaga atgaatagct
      181 tctgccagga tcctgactgg cgacgtaatc cacgaaaatc tatacattaa gacgatctct
      241 ttttccgtca tcgcctaggt ctagggcggc ggatttgtcc tactcaggag agcgttcacc
      301 gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga
      361 tgcctctaga tgatgcctgg ctctagtagt gatctacact agcactatca gtgttattaa
      421 tttttaaagt atgggcaatc aattgctcct gttaaaattg ctttagaaat actttggcag
      481 cggtttgttg tattgagttt catttgcgca ttggttaaat ggaaagtgac agtacgctca
      541 ctgcaaccta atatttttga aatatcccaa gagctttttc cttcgcatgc ccacgctaaa
      601 cattcttttt ctcttttggt taaatcgttg tttgatttat tatttgctat atttattttt
      661 cgataattat caactagaga aggaacaatt aatggtatgt tcatacacgc atgtaaaaat
      721 aaactatcta tatagttgtc tttttctgaa tgtgcaaaac taagcattcc gaagccattg
      781 ttagccgtat gaatagggaa actaaaccca gtgataagac ctgatgtttt cgcttcttta
      841 attacatttg gagatttttt atttacagca ttgttttcaa atatattcca attaattggt
      901 gaatgattgg agttagaata atctactata ggatcatatt ttattaaatt agcgtcatca
      961 taatattgcc tccatttttt agggtaatta tctaggattg aaatatcaga tttaaccata
     1021 gaatgaggat aaatgatcgc gagtaaataa tattcacaat gtaccatttt agtcatatca
     1081 gataagcatt gattaatatc attattgctt ctacaagctt taattttatt aattattctg
     1141 tatgtgtcgt cggcatttat gtttttcatg gtacctttct cctctttaat gaattcgcta
     1201 gcacagtacc taggactgag ctagctgtca actcgaggtg aagacgaaag ggcctcgtga
     1261 tacgcctatt tttataggtt aatgtcatga taataaagat cctggtaact cgagaattgt
     1321 gagcggataa caattgacat tgtgagcgga taacaagata ctgagcacat ctacctctgg
     1381 cggtgataga tgattacctc tggcggtgat agatgattac ctctggcggt gatagaattc
     1441 attaaagagg agaaaggtac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1501 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1561 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1621 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1681 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1741 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1801 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     1861 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1921 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1981 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2041 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2101 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2161 atggacgagc tgtacaagta aagcggccgc gactctagag gcatcaaata aaacgaaagg
     2221 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     2281 gtaggacaaa tccgccgccc tagacctagg aacaaatgcg agatcttgat gttggcgtct
     2341 tcgttgtcga ctgttcgtct tcgtcccagt agacgtcagg tggcactttt cggggaaatg
     2401 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2461 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2521 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2581 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2641 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     2701 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     2761 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     2821 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     2881 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     2941 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3001 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3061 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3121 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3181 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3241 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3301 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3361 attggtaact gtcagaccaa gtttacgagc tcgcttggac tcctgttgat agatccagta
     3421 atgacctcag aactccatct ggatttgttc agaacgctcg gttgccgccg ggcgtttttt
     3481 attggtgaga atccaagcac tagtggctgg cactctgtcg ataccccacc gagaccccat
     3541 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3601 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     3661 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3721 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3781 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3841 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3901 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3961 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4021 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4081 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4141 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4201 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4261 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4321 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4381 gggcggagcc tatggaaa
//
