LOCUS       sequence-252641-        4052 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing sfGFP under control of benzoate responsive promoter 
            (pBen) and expressing mRFP under constitutive promoter J23101, and 
            RBS B0032.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4052)
  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 4052)
  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-mRFP
FEATURES             Location/Qualifiers
     source          1..4052
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..55
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     CDS             83..760
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      776..833
                     /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(857..874)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1028..1616)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1108..1127)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1849..2664)
                     /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(1849..2664)
                     /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     2572..2591
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2656..2675
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2869..2887
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2891..2934)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             3192..3905
                     /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      3931..4002
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4018..4045
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 actagagatg accccatctg tttacagcta gctcagtcct aggtattatg ctagctagta
       61 gagtcacaca ggaaagtagt agatggcttc ctccgaagac gttatcaaag agttcatgcg
      121 tttcaaagtt cgtatggaag gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg
      181 tgaaggtcgt ccgtacgaag gtacccagac cgctaaactg aaagttacca aaggtggtcc
      241 gctgccgttc gcttgggaca tcctgtcccc gcagttccag tacggttcca aagcttacgt
      301 taaacacccg gctgacatcc cggactacct gaaactgtcc ttcccggaag gtttcaaatg
      361 ggaacgtgtt atgaacttcg aagacggtgg tgttgttacc gttacccagg actcctccct
      421 gcaagacggt gagttcatct acaaagttaa actgcgtggt accaacttcc cgtccgacgg
      481 tccggttatg cagaaaaaaa ccatgggttg ggaagcttcc accgaacgta tgtacccgga
      541 agacggtgct ctgaaaggtg aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta
      601 cgacgctgaa gttaaaacca cctacatggc taaaaaaccg gttcagctgc cgggtgctta
      661 caaaaccgac atcaaactgg acatcacctc ccacaacgaa gactacacca tcgttgaaca
      721 gtacgaacgt gctgaaggtc gtcactccac cggtgcttaa tgcgtcttcc tgcagtccgg
      781 caaaaaaggg caaggtgtca ccaccctgcc ctttttcttt aaaaccgaaa agattacttc
      841 gcgttatgca ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      901 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      961 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1021 tggcgttttt ccacaggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1081 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1141 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1201 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1261 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1321 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1381 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1441 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1501 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1561 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1621 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1681 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1741 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagctcg agtcccgtca
     1801 agtcagcgta atgctctgcc agtgttacaa ccaattaacc aattctgatt agaaaaactc
     1861 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     1921 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     1981 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     2041 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     2101 gaatggcaaa agcttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     2161 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     2221 acgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgaat gcaaccggcg
     2281 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     2341 ctggaatgct gttttcccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     2401 gataaaatgc ttgatggtcg gaagaggcat aaattccgtc agccagttta gtctgaccat
     2461 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     2521 atcgggcttc ccatacaatc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     2581 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tggagcaaga
     2641 cgtttcccgt tgaatatggc tcataacacc ccttgtatta ctgtttatgt aagcagacag
     2701 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     2761 cacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgaaggatc agctcgagtg
     2821 ccacttgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     2881 acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga tttctggaat
     2941 tcgaagagcc tcggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     3001 cgatcaacgt ctcattttcg ccagatatcg gacttgggcc tactgttcga agcattgcca
     3061 ttttctgaag ttaccgaaaa agtaccgaac atccgtaaat ctggataacg ttctgcacaa
     3121 tccggatagc cccccgccag ccgtctccct aacctgacca ggtctaaaca ataacaaggg
     3181 agagtctggc catgcgtaaa ggcgaagagc tgttcactgg tgtcgtccct attctggtgg
     3241 aactggatgg tgatgtcaac ggtcataagt tttccgtgcg tggcgagggt gaaggtgacg
     3301 caactaatgg taaactgacg ctgaagttca tctgtactac tggtaaactg ccggtacctt
     3361 ggccgactct ggtaacgacg ctgacttatg gtgttcagtg ctttgctcgt tatccggacc
     3421 atatgaagca gcatgacttc ttcaagtccg ccatgccgga aggctatgtg caggaacgca
     3481 cgatttcctt taaggatgac ggcacgtaca aaacgcgtgc ggaagtgaaa tttgaaggcg
     3541 ataccctggt aaaccgcatt gagctgaaag gcattgactt taaagaagac ggcaatatcc
     3601 tgggccataa gctggaatac aattttaaca gccacaatgt ttacatcacc gccgataaac
     3661 aaaaaaatgg cattaaagcg aattttaaaa ttcgccacaa cgtggaggat ggcagcgtgc
     3721 agctggctga tcactaccag caaaacactc caatcggtga tggtcctgtt ctgctgccag
     3781 acaatcacta tctgagcacg caaagcgttc tgtctaaaga tccgaacgag aaacgcgatc
     3841 atatggttct gctggagttc gtaaccgcag cgggcatcac gcatggtatg gatgaactgt
     3901 acaaatgatg ataatactag agccaggcat caaataaaac gaaaggctca gtcgaaagac
     3961 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
     4021 tcaccttcgg gtgggccttt ctgcgtttat at
//
