LOCUS       sequence-225523-        4715 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ""Strong constitutive expression of metabolic enzyme, CocE, for 
            cell-free expression"".
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4715)
  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 4715)
  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-CocE
FEATURES             Location/Qualifiers
     source          1..4715
                     /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"
     terminator      2189..2260
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2276..2303
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2530..2547)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2701..3289)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2781..2800)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3460..4251,4252..4320))
                     /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     4083..4102
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4321..4425)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4493..4511
                     /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 agatgatggt tgacggtaac tactctgttg cttctaacgt tatggttccg atgcgtgacg
      421 gtgttcgtct ggctgttgac ctgtaccgtc cggacgctga cggtccggtt ccggttctgc
      481 tggttcgtaa cccgtacgac aaattcgacg ttttcgcttg gtctacccag tctaccaact
      541 ggctggaatt tgttcgtgac ggttacgctg ttgttatcca ggacacccgt ggtctgttcg
      601 cttctgaagg tgaatttgtt ccgcacgttg acgacgaagc tgacgctgaa gacaccctgt
      661 cttggatttt ggaacaggct tggtgcgacg gtaacgttgg tatgttcggt gtttcttacc
      721 tgggtgttac ccagtggcag gctgctgttt ctggtgttgg tggtctgaaa gctatcgctc
      781 cgtctatggc ttctgctgac ctgtaccgtg ctccgtggta cggtccgggt ggtgctctgt
      841 ctgttgaagc gctgctgggt tggtctgctc tgatcggtac cggtctgatc acctctcgtt
      901 ctgacgctcg tccggaagac gctgctgact tcgttcagct ggctgctatc ctgaacgacg
      961 ttgctggtgc tgcttctgtt accccgctgg ctgaacagcc gctgctgggt cgtctgatcc
     1021 cgtgggttat cgaccaggtt gttgaccacc cggacaacga cgaatcttgg cagtctatct
     1081 ctctgttcga acgtctgggt ggtctggcta ccccggctct gatcaccgct ggttggtacg
     1141 acggtttcgt tggtgaaagc ctgcgtacct tcgttgctgt taaagacaac gctgacgctc
     1201 gtctggttgt tggtccgtgg tcccactcta acctgaccgg tcgtaacgct gaccgtaaat
     1261 tcggtatcgc tgctacctac ccgatccagg aagctaccac catgcacaaa gctttcttcg
     1321 accgtcacct gcgtggtgaa accgacgcac ttgctggtgt tccgaaagtt cgtctgttcg
     1381 ttatgggtat cgacgaatgg cgtgacgaaa ccgactggcc gctgccggac accgcttaca
     1441 ccccgttcta cctgggtggt tctggtgctg ctaacacctc taccggtggt ggtaccctgt
     1501 ctacctctat cagcggtacc gaatctgctg acacctacct gtacgacccg gctgacccgg
     1561 ttccgagcct gggtggtacc ctgctgttcc acaacggtga caacggtccg gctgaccagc
     1621 gtccgatcca cgaccgtgac gacgttctgt gctactctac cgaagttctg accgacccgg
     1681 ttgaagttac cggtaccgtt tctgctcgtc tgttcgtttc ttcttctgct gttgacaccg
     1741 acttcaccgc taaactggtt gacgttttcc cggacggtcg tgctatcgct ctgtgcgacg
     1801 gtatcgttcg tatgcgttac cgtgaaaccc tggttaaccc gaccctgatc gaagctggtg
     1861 aaatctacga agttgctatc gacatgctgg ctacctctaa cgttttcctg ccgggtcacc
     1921 gtatcatggt tcaggtttct tcttctaact tcccgaaata cgaccgtaac tctaacaccg
     1981 gtggtgttat cgctcgtgaa cagctggaag aaatgtgcac cgctgttaac cgtatccacc
     2041 gtggtccgga acacccgagc cacatcgttc tgccgatcat caaacgttaa gcttacttta
     2101 tctgagaata gtcaatcttc ggaaatccca ggtggcatgc taaaagtctc gtaaagcgtt
     2161 ctatcaataa cccgttggtg ccaggcatca aataaaacga aaggctcagt cgaaagactg
     2221 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc acactggctc
     2281 accttcgggt gggcctttct gcgtttatac cgtctcagaa tcggccgtga acaataaaat
     2341 agtttcggta ttattgacca cttccgagta gaatcgtgct tcagtaagag tcgaccgatg
     2401 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc
     2461 gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc
     2521 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2581 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2641 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2701 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2761 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2821 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2881 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2941 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     3001 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     3061 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     3121 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     3181 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     3241 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     3301 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     3361 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     3421 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3481 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3541 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3601 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3661 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3721 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3781 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3841 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3901 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3961 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     4021 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg
     4081 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     4141 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     4201 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     4261 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     4321 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     4381 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     4441 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4501 tatcacgagg ccctttcgtc ttcaagaatt ctggcgaatc ctctgaccag ccagaaaacg
     4561 acctttctgt ggtgaaaccg gatgctgcaa ttcagagcgc cagcaagtgg gggacagcag
     4621 aagacctgac cgccgcagag tggatgtttg acatggtgaa gactatcgca ccatcagcca
     4681 gaaaaccgaa ttttgctggg tgggctaacg atatc
//
