LOCUS       sequence-225522-        3950 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ""Strong constitutive expression of transcription factor, BenR, for 
            cell-free expression"".
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3950)
  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 3950)
  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-BenR
FEATURES             Location/Qualifiers
     source          1..3950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      148..175
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1424..1495
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1511..1538
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1765..1782)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1936..2524)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2016..2035)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2695..3486,3487..3555))
                     /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     3318..3337
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3556..3660)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3728..3746
                     /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 tcttatgagc taacaccgtg cgtgttgaca
      301 attttacctc tggcggtgat aatggttgca gctagcaata attttgttta actttaagaa
      361 gagatatgat ggaatctcgt ctgctgtctg aacgttcttc tgttttccac cacgctgacc
      421 cgtacgctgt ttctgactac gttaaccagc acgttggtca gcactgcatc ggtctgtctc
      481 gtaccaccca cccgcaggct tctctgtctc accgtaaatt cgctgaactg gacctgtgcc
      541 gtatctctta cggtggttct gttcgtgtta cctctccggc tctggaaacc atctaccacc
      601 tgcaggttct gctgaacggt aactgcctgt ggcgtggtca caaacgtgaa cagcacctgg
      661 ttccgggtga actgctgctg atcaacccgg acgacccggt tgacctgacc tactctgaag
      721 actgcgaaaa attcatcctg aaagttccga cccgtctgct ggactctatc tgcgacgaac
      781 agcgttggca gcgtccggac ggtggtgttc gtttcctgcg taaccactac cgtctggacg
      841 aactggacgg tttcgttaac ctgctggcta tggtttgcca cgaagctgaa gtttctgact
      901 ctctgccgcg tgttcagggt cactactctc agatcgttgc ttctaaactg ctgaccctga
      961 tgtctaccaa catccgtcgt gaatctctgt ctgctccgca ggctggtctg gaacgtatcc
     1021 tggactacat cgaacgtaac ctgaaactgg aactgtctgc tgaagttctg gctgaacagg
     1081 cttgcatgtc tctgcgttct ctgtacgctc tgttcgacca gcacctgggt atcaccccga
     1141 aacactacgt tcgtcagcgt aaactggaac gtgttcacgc ttgcctgtct gacccgacct
     1201 gcggtgttcg ttctgttacc gaactggctc tggactacgg tttcctgcac ctgggtcgtt
     1261 tctctgaaat ctaccgtcag cagttcggtg aactgccgtc tcagaccttc aaacgtcgtg
     1321 cttaagctta ctttatctga gaatagtcaa tcttcggaaa tcccaggtgg catgctaaaa
     1381 gtctcgtaaa gcgttctatc aataacccgt tggtgccagg catcaaataa aacgaaaggc
     1441 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
     1501 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tataccgtct cagaatcggc
     1561 cgtgaacaat aaaatagttt cggtattatt gaccacttcc gagtagaatc gtgcttcagt
     1621 aagagtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     1681 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     1741 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1801 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1861 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1921 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1981 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2041 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2101 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2161 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2221 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2281 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2341 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2401 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2461 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2521 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2581 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2641 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2701 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2761 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2821 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2881 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2941 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3001 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3061 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3121 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3181 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3241 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3301 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3361 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3421 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3481 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3541 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3601 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3661 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     3721 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattctggc gaatcctctg
     3781 accagccaga aaacgacctt tctgtggtga aaccggatgc tgcaattcag agcgccagca
     3841 agtgggggac agcagaagac ctgaccgccg cagagtggat gtttgacatg gtgaagacta
     3901 tcgcaccatc agccagaaaa ccgaattttg ctgggtgggc taacgatatc
//
