LOCUS       sequence-234180-        4686 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 4686)
  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 4686)
  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..4686
                     /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(2501..2518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2672..3260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2752..2771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3431..4222,4223..4291))
                     /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     4054..4073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4292..4396)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4464..4482
                     /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 gcaacgaaag gctactgtct tctttatcat
     2461 gcaactcgta ggacaggtgc cggcagcgct cttccgcttc ctcgctcact gactcgctgc
     2521 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2581 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2641 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2701 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2761 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2821 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta
     2881 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2941 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3001 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3061 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3121 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3181 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3241 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3301 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3361 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3421 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3481 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3541 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3601 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3661 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3721 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3781 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3841 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3901 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3961 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4021 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4081 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4141 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4201 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4261 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4321 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4381 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     4441 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt cttcaagaat
     4501 tctggcgaat cctctgacca gccagaaaac gacctttctg tggtgaaacc ggatgctgca
     4561 attcagagcg gcagcaagtg ggggacagca gaagacctga ccgccgcaga gtggatgttt
     4621 gacatggtga agactatcgc accatcagcc agaaaaccga attttgctgg gtgggctaac
     4681 gatatc
//
