LOCUS       sequence-247142-        3611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Strong constitutive expression of super-folder GFP for cell-free 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3611)
  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 3611)
  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: pBEAST2-sfGFP
FEATURES             Location/Qualifiers
     source          1..3611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      148..175
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     RBS             343..365
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             370..1083
                     /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"
     primer_bind     complement(1426..1443)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1597..2185)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1677..1696)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2356..3147,3148..3216))
                     /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     2979..2998
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3217..3321)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3389..3407
                     /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 ggagatataa tgcgtaaagg cgaagagctg ttcactggtg tcgtccctat tctggtggaa
      421 ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga aggtgacgca
      481 actaatggta aactgacgct gaagttcatc tgtactactg gtaaactgcc ggtaccttgg
      541 ccgactctgg taacgacgct gacttatggt gttcagtgct ttgctcgtta tccggaccat
      601 atgaagcagc atgacttctt caagtccgcc atgccggaag gctatgtgca ggaacgcacg
      661 atttccttta aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat
      721 accctggtaa accgcattga gctgaaaggc attgacttta aagaagacgg caatatcctg
      781 ggccataagc tggaatacaa ttttaacagc cacaatgttt acatcaccgc cgataaacaa
      841 aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag
      901 ctggctgatc actaccagca aaacactcca atcggtgatg gtcctgttct gctgccagac
      961 aatcactatc tgagcacgca aagcgttctg tctaaagatc cgaacgagaa acgcgatcat
     1021 atggttctgc tggagttcgt aaccgcagcg ggcatcacgc atggtatgga tgaactgtac
     1081 aaatgatgaa ctttatctga gaatagtcaa tcttcggaaa tcccaggtgg catgctaaaa
     1141 gtctcgtaaa gcgttctatc aataacccgt tggtgcaaag cccgccgaaa ggcgggcttt
     1201 tctgtccgtc tcagaatcgg ccgtgaacaa taaaatagtt tcggtattat tgaccacttc
     1261 cgagtagaat cgtgcttcag taagagtcga ccgatgccct tgagagcctt caacccagtc
     1321 agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt
     1381 atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc tcactgactc
     1441 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     1501 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1561 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1621 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1681 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1741 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1801 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1861 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1921 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1981 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2041 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2101 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2161 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2221 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     2281 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     2341 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     2401 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     2461 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     2521 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2581 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2641 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     2701 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2761 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2821 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2881 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2941 gcggcgaccg agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag
     3001 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3061 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3121 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3181 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3241 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     3301 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     3361 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     3421 agaattctgg cgaatcctct gaccagccag aaaacgacct ttctgtggtg aaaccggatg
     3481 ctgcaattca gagcgccagc aagtggggga cagcagaaga cctgaccgcc gcagagtgga
     3541 tgtttgacat ggtgaagact atcgcaccat cagccagaaa accgaatttt gctgggtggg
     3601 ctaacgatat c
//
