LOCUS       sequence-225526-        3710 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 3710)
  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 3710)
  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-J23101-sfGFP
FEATURES             Location/Qualifiers
     source          1..3710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      148..175
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        301..335
                     /label=BBa_J23101
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.7"
     CDS             363..1076
                     /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      1184..1255
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1271..1298
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1525..1542)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1696..2284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1776..1795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2455..3246,3247..3315))
                     /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     3078..3097
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3316..3420)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3488..3506
                     /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 tcttaggata ctagaggatg accccatctg
      301 tttacagcta gctcagtcct aggtattatg ctagctagta gagtcacaca ggaaagtagt
      361 agatgcgtaa aggcgaagag ctgttcactg gtgtcgtccc tattctggtg gaactggatg
      421 gtgatgtcaa cggtcataag ttttccgtgc gtggcgaggg tgaaggtgac gcaactaatg
      481 gtaaactgac gctgaagttc atctgtacta ctggtaaact gccggtacct tggccgactc
      541 tggtaacgac gctgacttat ggtgttcagt gctttgctcg ttatccggac catatgaagc
      601 agcatgactt cttcaagtcc gccatgccgg aaggctatgt gcaggaacgc acgatttcct
      661 ttaaggatga cggcacgtac aaaacgcgtg cggaagtgaa atttgaaggc gataccctgg
      721 taaaccgcat tgagctgaaa ggcattgact ttaaagaaga cggcaatatc ctgggccata
      781 agctggaata caattttaac agccacaatg tttacatcac cgccgataaa caaaaaaatg
      841 gcattaaagc gaattttaaa attcgccaca acgtggagga tggcagcgtg cagctggctg
      901 atcactacca gcaaaacact ccaatcggtg atggtcctgt tctgctgcca gacaatcact
      961 atctgagcac gcaaagcgtt ctgtctaaag atccgaacga gaaacgcgat catatggttc
     1021 tgctggagtt cgtaaccgca gcgggcatca cgcatggtat ggatgaactg tacaaatgat
     1081 gataagctta ctttatctga gaatagtcaa tcttcggaaa tcccaggtgg catgctaaaa
     1141 gtctcgtaaa gcgttctatc aataacccgt tggtgccagg catcaaataa aacgaaaggc
     1201 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
     1261 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tataccgtct cagaatcggc
     1321 cgtgaacaat aaaatagttt cggtattatt gaccacttcc gagtagaatc gtgcttcagt
     1381 aagagtcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
     1441 ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact cgtaggacag
     1501 gtgccggcag cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1561 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1621 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1681 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1741 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1801 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1861 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1921 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1981 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2041 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2101 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2161 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2221 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2281 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2341 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2401 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2461 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2521 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     2581 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     2641 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     2701 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     2761 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     2821 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     2881 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     2941 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     3001 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     3061 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     3121 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     3181 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     3241 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     3301 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     3361 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     3421 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     3481 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa gaattctggc gaatcctctg
     3541 accagccaga aaacgacctt tctgtggtga aaccggatgc tgcaattcag agcgccagca
     3601 agtgggggac agcagaagac ctgaccgccg cagagtggat gtttgacatg gtgaagacta
     3661 tcgcaccatc agccagaaaa ccgaattttg ctgggtgggc taacgatatc
//
