LOCUS       sequence-225646-        3676 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Strong constitutive expression of metabolic enzyme, HipO, for 
            cell-free expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3676)
  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 3676)
  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: pBEST-HipO
FEATURES             Location/Qualifiers
     source          1..3676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             7..29
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1364..1381)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1535..2123)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1615..1634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2294..3085,3086..3154))
                     /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     2917..2936
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3155..3259)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3327..3345
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aataattttg tttaacttta agaaggagat ataccatgaa cctgatcccg gaaatcctgg
       61 acctgcaggg tgaattcgaa aaaatccgtc accagatcca cgaaaacccg gaactgggtt
      121 tcgacgaact gtgcaccgct aaactggttg ctcagaaact gaaagaattc ggttacgaag
      181 tttacgaaga aatcggtaaa accggtgttg ttggtgttct gaaaaaaggt aactctgaca
      241 aaaaaatcgg tctgcgtgct gacatggacg ctctgccgct gcaggaatgc accaacctgc
      301 cgtacaaatc taaaaaagaa aacgttatgc acgcttgcgg tcacgacggt cacaccacct
      361 ctctgctgct ggctgctaaa tacctggctt ctcagaactt caacggtgct ctgaacctgt
      421 acttccagcc ggctgaagaa ggtctgggtg gtgctaaagc tatgatcgaa gacggtctgt
      481 tcgaaaaatt cgactctgac tacgttttcg gttggcacaa catgccgttc ggttctgaca
      541 aaaaattcta cctgaaaaaa ggtgctatga tggcttcttc tgactcttac tctatcgaag
      601 ttatcggtcg tggtggtcac ggttctgctc cggaaaaagc taaagacccg atctacgctg
      661 cttctctgct gatcgttgct ctgcagtcta tcgtttctcg taacgttgac ccgcagaact
      721 ctgctgttgt ttctatcggt gctttcaacg ctggtcacgc tttcaacatc atcccggaca
      781 tcgctaccat caaaatgtct gttcgtgctc tggacaacga aacccgtaaa ctgaccgaag
      841 aaaaaatcta caaaatctgc aaaggtatcg ctcaggctaa cgacatcgaa atcaaaatca
      901 acaaaaacgt tgttgctccg gttaccatga acaacgacga agctgttgac ttcgcttctg
      961 aagttgctaa agaactgttc ggtgaaaaaa actgcgaatt caaccaccgt ccgctgatgg
     1021 cttctgaaga cttcggtttc ttctgcgaaa tgaaaaaatg cgcttacgct ttcctggaaa
     1081 acgaaaacga catctacctg cacaactctt cttacgtttt caacgacaaa ctgctggctc
     1141 gtgctgcttc ttactacgct aaactggctc tgaaatacct gaaataactc gagcaaagcc
     1201 cgccgaaagg cgggcttttc tgtgtcgacc gatgcccttg agagccttca acccagtcag
     1261 ctccttccgg tgggcgcggg gcatgactat cgtcgccgca cttatgactg tcttctttat
     1321 catgcaactc gtaggacagg tgccggcagc gctcttccgc ttcctcgctc actgactcgc
     1381 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     1441 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     1501 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1561 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1621 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1681 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1741 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1801 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1861 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1921 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1981 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2041 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2101 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     2161 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     2221 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     2281 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2341 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2401 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     2461 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2521 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     2581 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     2641 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     2701 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2761 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2821 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2881 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     2941 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3001 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3061 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3121 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3181 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3241 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     3301 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag
     3361 aattctggcg aatcctctga ccagccagaa aacgaccttt ctgtggtgaa accggatgct
     3421 gcaattcaga gcgccagcaa gtgggggaca gcagaagacc tgaccgccgc agagtggatg
     3481 tttgacatgg tgaagactat cgcaccatca gccagaaaac cgaattttgc tgggtgggct
     3541 aacgatatcc gcctgatgcg tgaacgtgac ggacgtaacc accgcgacat gtgtgtgctg
     3601 ttccgctggg catgctgagc taacaccgtg cgtgttgaca attttacctc tggcggtgat
     3661 aatggttgca gctagc
//
