LOCUS       sequence-252642-        4345 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing sfGFP under control of benzoate responsive promoter 
            (pBen) and expressing BenR transcription factor gene under 
            constitutive promoter J23101, and RBS B0032.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4345)
  AUTHORS   Pandi A, Koch M, Voyvodic PL, Soudier P, Bonnet J, Kushwaha M, 
            Faulon JL
  TITLE     Metabolic perceptrons for neural computing in biological systems.
  JOURNAL   Nat Commun. 2019 Aug 28;10(1):3880. doi: 10.1038/s41467-019-11889-0.
  PUBMED    31462649
REFERENCE   2  (bases 1 to 4345)
  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"; journalName: "Nat 
            Commun."; date: "2019-08-28"; volume: "10(1):3880. doi"; pages: " 
            10.1038/s41467-019-11889-0"
COMMENT     Sequence Label: pSB1K3_pBen-sfGFP_J23101-BenR
FEATURES             Location/Qualifiers
     source          1..4345
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(272..1087)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(272..1087)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     995..1014
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1079..1098
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1292..1310
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1314..1357)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             1615..2328
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      2354..2425
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2441..2468
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2496..2530
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     terminator      3544..3601
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(3625..3642)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3796..4345,1..39))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3876..3895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
       61 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
      121 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
      181 agtatatatg agtaaacttg gtctgacagc tcgagtcccg tcaagtcagc gtaatgctct
      241 gccagtgtta caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa
      301 actgcaattt attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta
      361 atgaaggaga aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg
      421 cgattccgac tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt
      481 tatcaagtga gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat
      541 gcatttcttt ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg
      601 catcaaccaa accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc
      661 tgttaaaagg acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg
      721 catcaacaat attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc
      781 cggggatcgc agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg
      841 tcggaagagg cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat
      901 tggcaacgct acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca
      961 atcgatagat tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata
     1021 aatcagcatc catgttggaa tttaatcgcg gcctggagca agacgtttcc cgttgaatat
     1081 ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg
     1141 atatattttt atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt
     1201 gaataaatcg aacttttgct gagttgaagg atcagctcga gtgccacttg acgtctaaga
     1261 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc agaatttcag
     1321 ataaaaaaaa tccttagctt tcgctaagga tgatttctgg aattcgaaga gcctcggtag
     1381 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
     1441 tcgccagata tcggacttgg gcctactgtt cgaagcattg ccattttctg aagttaccga
     1501 aaaagtaccg aacatccgta aatctggata acgttctgca caatccggat agccccccgc
     1561 cagccgtctc cctaacctga ccaggtctaa acaataacaa gggagagtct ggccatgcgt
     1621 aaaggcgaag agctgttcac tggtgtcgtc cctattctgg tggaactgga tggtgatgtc
     1681 aacggtcata agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg
     1741 acgctgaagt tcatctgtac tactggtaaa ctgccggtac cttggccgac tctggtaacg
     1801 acgctgactt atggtgttca gtgctttgct cgttatccgg accatatgaa gcagcatgac
     1861 ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc ctttaaggat
     1921 gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc
     1981 attgagctga aaggcattga ctttaaagaa gacggcaata tcctgggcca taagctggaa
     2041 tacaatttta acagccacaa tgtttacatc accgccgata aacaaaaaaa tggcattaaa
     2101 gcgaatttta aaattcgcca caacgtggag gatggcagcg tgcagctggc tgatcactac
     2161 cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc cagacaatca ctatctgagc
     2221 acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg atcatatggt tctgctggag
     2281 ttcgtaaccg cagcgggcat cacgcatggt atggatgaac tgtacaaatg atgataatac
     2341 tagagccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     2401 ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc
     2461 tttctgcgtt tatatactag agatgacccc atctgtttac agctagctca gtcctaggta
     2521 ttatgctagc tagtagagtc acacaggaaa gtagtagatg gaatctcgtc tgctgtctga
     2581 acgttcttct gttttccacc acgctgaccc gtacgctgtt tctgactacg ttaaccagca
     2641 cgttggtcag cactgcatcg gtctgtctcg taccacccac ccgcaggctt ctctgtctca
     2701 ccgtaaattc gctgaactgg acctgtgccg tatctcttac ggtggttctg ttcgtgttac
     2761 ctctccggct ctggaaacca tctaccacct gcaggttctg ctgaacggta actgcctgtg
     2821 gcgtggtcac aaacgtgaac agcacctggt tccgggtgaa ctgctgctga tcaacccgga
     2881 cgacccggtt gacctgacct actctgaaga ctgcgaaaaa ttcatcctga aagttccgac
     2941 ccgtctgctg gactctatct gcgacgaaca gcgttggcag cgtccggacg gtggtgttcg
     3001 tttcctgcgt aaccactacc gtctggacga actggacggt ttcgttaacc tgctggctat
     3061 ggtttgccac gaagctgaag tttctgactc tctgccgcgt gttcagggtc actactctca
     3121 gatcgttgct tctaaactgc tgaccctgat gtctaccaac atccgtcgtg aatctctgtc
     3181 tgctccgcag gctggtctgg aacgtatcct ggactacatc gaacgtaacc tgaaactgga
     3241 actgtctgct gaagttctgg ctgaacaggc ttgcatgtct ctgcgttctc tgtacgctct
     3301 gttcgaccag cacctgggta tcaccccgaa acactacgtt cgtcagcgta aactggaacg
     3361 tgttcacgct tgcctgtctg acccgacctg cggtgttcgt tctgttaccg aactggctct
     3421 ggactacggt ttcctgcacc tgggtcgttt ctctgaaatc taccgtcagc agttcggtga
     3481 actgccgtct cagaccttca aacgtcgtgc ttaagcttcg gtgcttaatg cgtcttcctg
     3541 cagtccggca aaaaagggca aggtgtcacc accctgccct ttttctttaa aaccgaaaag
     3601 attacttcgc gttatgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     3661 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3721 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3781 cgcgttgctg gcgtttttcc acaggctccg cccccctgac gagcatcaca aaaatcgacg
     3841 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     3901 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     3961 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4021 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4081 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4141 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4201 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     4261 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4321 cgctggtagc ggtggttttt ttgtt
//
