LOCUS       sequence-237550-        4699 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Output expression of firefly luciferase under the activation of BenR
            transcription factor for cell-free expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4699)
  AUTHORS   Voyvodic PL, Pandi A, Koch M, Conejero I, Valjent E, Courtet P, 
            Renard E, Faulon JL, Bonnet J
  TITLE     Plug-and-play metabolic transducers expand the chemical detection 
            space of cell-free biosensors.
  JOURNAL   Nat Commun. 2019 Apr 12;10(1):1697. doi: 10.1038/s41467-019-09722-9.
  PUBMED    30979906
REFERENCE   2  (bases 1 to 4699)
  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: "2019-04-12"; volume: "10(1):1697. doi"; pages: " 
            10.1038/s41467-019-09722-9"
COMMENT     Sequence Label: pBEAST-pBen-Luc
FEATURES             Location/Qualifiers
     source          1..4699
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      148..175
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             426..2078
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(492..510)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     terminator      2173..2244
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2260..2287
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2514..2531)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2685..3273)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2765..2784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3444..4235,4236..4304))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4067..4086
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4305..4409)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4477..4495
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgcctgatgc gtgaacgtga cggacgtaac caccgcgaca tgtgtgtgct gttccgctgg
       61 ctcggatacc cttactctgt tgaaaacgaa tagataggtt aaggaacggt tatttctgcg
      121 tagatctatc ttacacagca tcacactggc tcaccttcgg gtgggccttt ctgcgtttat
      181 atactagaga gagaatataa aaagccagat tattaatccg gcttttttat tatttaggca
      241 actgaaacga ttcggatcct gtattactat tcttaactgt tcgaagcatt gccattttct
      301 gaagttaccg aaaaagtacc gaacatccgt aaatctggat aacgttctgc acaatccgga
      361 tagccccccg ccagccgtct ccctaacctg accaggtcta aacaataaca agggagagtc
      421 tggccatgga agacgccaaa aacataaaga aaggcccggc gccattctat ccgctggaag
      481 atggaaccgc tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa
      541 caattgcttt tacagatgca catatcgagg tggacatcac ttacgctgag tacttcgaaa
      601 tgtccgttcg gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg
      661 tcgtatgcag tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg
      721 gagttgcagt tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatgg
      781 gcatttcgca gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg
      841 tgcaaaaaaa gctcccaatc atccaaaaaa ttattatcat ggattctaaa acggattacc
      901 agggatttca gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat
      961 acgattttgt gccagagtcc ttcgataggg acaagacaat tgcactgatc atgaactcct
     1021 ctggatctac tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat
     1081 tctcgcatgc cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa
     1141 gtgttgttcc attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg
     1201 gatttcgagt cgtcttaatg tatagatttg aagaagagct gtttctgagg agccttcagg
     1261 attacaagat tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc gccaaaagca
     1321 ctctgattga caaatacgat ttatctaatt tacacgaaat tgcttctggt ggcgctcccc
     1381 tctctaagga agtcggggaa gcggttgcca agaggttcca tctgccaggt atcaggcaag
     1441 gatatgggct cactgagact acatcagcta ttctgattac acccgagggg gatgataaac
     1501 cgggcgcggt cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg
     1561 ggaaaacgct gggcgttaat caaagaggcg aactgtgtgt gagaggtcct atgattatgt
     1621 ccggttatgt aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac
     1681 attctggaga catagcttac tgggacgaag acgaacactt cttcatcgtt gaccgcctga
     1741 agtctctgat taagtacaaa ggctatcagg tggctcccgc tgaattggaa tccatcttgc
     1801 tccaacaccc caacatcttc gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg
     1861 aacttcccgc cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg
     1921 tggattacgt cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg
     1981 tggacgaagt accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc
     2041 tcataaaggc caagaagggc ggaaagatcg ccgtgtaaac tttatctgag aatagtcaat
     2101 cttcggaaat cccaggtggc atgctaaaag tctcgtaaag cgttctatca ataacccgtt
     2161 ggtgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     2221 tgttgtttgt cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct
     2281 ttctgcgttt ataccgtctc agaatcggcc gtgaacaata aaatagtttc ggtattattg
     2341 accacttccg agtagaatcg tgcttcagta agagtcgacc gatgcccttg agagccttca
     2401 acccagtcag ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg
     2461 tcttctttat catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc
     2521 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2581 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2641 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2701 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2761 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2821 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2881 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2941 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3001 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3061 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3121 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3181 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3241 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3301 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3361 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3421 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3481 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3541 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3601 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3661 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3721 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3781 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3841 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3901 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3961 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4021 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     4081 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4141 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4201 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4261 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4321 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4381 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     4441 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     4501 cgtcttcaag aattctggcg aatcctctga ccagccagaa aacgaccttt ctgtggtgaa
     4561 accggatgct gcaattcaga gcgccagcaa gtgggggaca gcagaagacc tgaccgccgc
     4621 agagtggatg tttgacatgg tgaagactat cgcaccatca gccagaaaac cgaattttgc
     4681 tgggtgggct aacgatatc
//
