LOCUS       sequence-225515-        3775 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ""Output expression of sfGFP 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 3775)
  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 3775)
  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-sfGFP
FEATURES             Location/Qualifiers
     source          1..3775
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      148..175
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             428..1141
                     /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      1249..1320
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1336..1363
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1590..1607)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1761..2349)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1841..1860)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2520..3311,3312..3380))
                     /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     3143..3162
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3381..3485)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3553..3571
                     /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 tcttaggact gttcgaagca ttgccatttt
      301 ctgaagttac cgaaaaagta ccgaacatcc gtaaatctgg ataacgttct gcacaatccg
      361 gatagccccc cgccagccgt ctccctaacc tgaccaggtc taaacaataa caagggagag
      421 tctggccatg cgtaaaggcg aagagctgtt cactggtgtc gtccctattc tggtggaact
      481 ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc gagggtgaag gtgacgcaac
      541 taatggtaaa ctgacgctga agttcatctg tactactggt aaactgccgg taccttggcc
      601 gactctggta acgacgctga cttatggtgt tcagtgcttt gctcgttatc cggaccatat
      661 gaagcagcat gacttcttca agtccgccat gccggaaggc tatgtgcagg aacgcacgat
      721 ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa gtgaaatttg aaggcgatac
      781 cctggtaaac cgcattgagc tgaaaggcat tgactttaaa gaagacggca atatcctggg
      841 ccataagctg gaatacaatt ttaacagcca caatgtttac atcaccgccg ataaacaaaa
      901 aaatggcatt aaagcgaatt ttaaaattcg ccacaacgtg gaggatggca gcgtgcagct
      961 ggctgatcac taccagcaaa acactccaat cggtgatggt cctgttctgc tgccagacaa
     1021 tcactatctg agcacgcaaa gcgttctgtc taaagatccg aacgagaaac gcgatcatat
     1081 ggttctgctg gagttcgtaa ccgcagcggg catcacgcat ggtatggatg aactgtacaa
     1141 atgatgataa gcttacttta tctgagaata gtcaatcttc ggaaatccca ggtggcatgc
     1201 taaaagtctc gtaaagcgtt ctatcaataa cccgttggtg ccaggcatca aataaaacga
     1261 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     1321 tactagagtc acactggctc accttcgggt gggcctttct gcgtttatac cgtctcagaa
     1381 tcggccgtga acaataaaat agtttcggta ttattgacca cttccgagta gaatcgtgct
     1441 tcagtaagag tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg
     1501 cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag
     1561 gacaggtgcc ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1621 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1681 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1741 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1801 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1861 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1921 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     1981 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2041 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2101 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2161 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2221 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2281 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2341 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2401 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2461 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2521 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2581 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2641 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2701 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2761 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2821 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2881 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     2941 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3001 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3061 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3121 tcttgcccgg cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3181 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3241 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3301 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3361 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3421 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3481 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     3541 ttaacctata aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt ctggcgaatc
     3601 ctctgaccag ccagaaaacg acctttctgt ggtgaaaccg gatgctgcaa ttcagagcgc
     3661 cagcaagtgg gggacagcag aagacctgac cgccgcagag tggatgtttg acatggtgaa
     3721 gactatcgca ccatcagcca gaaaaccgaa ttttgctggg tgggctaacg atatc
//
