LOCUS       sequence-430436-        4534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of HyPerFLEX.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4534)
  AUTHORS   Potekhina ES, Bass DI, Ezerina D, Fleckenstein DD, Chebotarev AS, 
            Sysoeva VA, Maltsev DI, Pak VV, Moshchenko AA, Sokolov AI, 
            Myasnyanko IN, Baranov MS, Fedotov AB, Zheltikov AM, Lanin AA, 
            Messens J, Belousov VV
  TITLE     A color-tailored fluorogenic sensor for hydrogen peroxide.
  JOURNAL   Nat Chem Biol. 2025 Oct 16. doi: 10.1038/s41589-025-02036-6.
  PUBMED    41102408
REFERENCE   2  (bases 1 to 4534)
  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 Chem 
            Biol. 2025 Oct 16. doi: 10.1038/s41589-025-02036-6."
COMMENT     Sequence Label: pQE30-HyPerFLEX
FEATURES             Location/Qualifiers
     source          1..4534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..54)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    62..78
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             127..144
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1282..1376
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1420..2079
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1486..1505)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2144..2215
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2276..2298)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2384..2524
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2439..2458
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2539..2556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2710..3298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2790..2809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3469..4260,4261..4329))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4092..4111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4330..4434)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4502..4520
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga
      121 ggatcgcatc accatcacca tcacggatcc cgggaattcg ccaccatgca cctggccaat
      181 gaggagcaaa acgaattgga aggtgctttt aagctgggct tgatctttac cgtcgccccg
      241 tacttgctgc ctaagttgat cgtcagcctg cgccgcaccg ccccgaagat gcctttgatg
      301 ttggaggaga attacaccca cacccttacc gagtcactga aacgcggcga cgttgacgcc
      361 atcattgtgg cagagccgtt tcaagagccg ggtattgtga ctgaaccttt gtatgacgag
      421 ccgtttttcg ttatcgttcc gaagggccat agcttcgagg agttggatgc cgttagccct
      481 cgcatgttgg gcgaagaaca agttctgctt ctgaccgagg gcaactgtat gcgtgaccag
      541 gtgcttagca tttttaccaa ggtcaaggtg cacatgaaga aagccctttc cggtgacagc
      601 tattgggtct ttgtgaaacg ggtaggtggc actggcggtg agcatgttgt ctttggcagt
      661 gaggacatcg agaacactat ggccaaaatg gacgacggac aactggatgg gttggccttt
      721 ggcgcaattc agctcgatgg tgacgggaat atcctgcagt acaatgctgc tgaaggagac
      781 atcacaggca gagatcccaa acaggtgatt gggaagaact tcttcaagga tgttgcacct
      841 ggaacggatt ctcccgagtt ttacggcaaa ttcaaggaag gcgtagcgtc agggaatctg
      901 aacaccatgt tcgaatggat gataccgact cttaccgatc atagttgcag cgagcttgcc
      961 gccaaacagc gcattcaggg tctcaccaac acccttcaag gctcaagcat cgataccatc
     1021 cgccacatgg ttgccagtgg cttggcaatc agcgtcttgc ctgccactgc ccttaccgaa
     1081 aacgaccaca tgctgtttag cattatcccg tttgaaggca ccccgcctag ccgccgcgtt
     1141 gtcctggcct atcgccgcaa ctttgtccgc cctaaggccc tttcagccat gaaagccgca
     1201 attatgcaaa gccaactgca tggtgttagc ttcatctgcg attgactcga ggccgactgt
     1261 aagcttaatt agctgagctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
     1321 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa
     1381 gctagcttgg cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     1441 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     1501 gttgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     1561 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     1621 aatgctcatc cggaatttcg tatggcaatg aaagacggtg agctggtgat atgggatagt
     1681 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     1741 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     1801 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     1861 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     1921 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     1981 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     2041 ttacaacagt actgcgatga gtggcagggc ggggcgtaat ttttttaagg cagttattgg
     2101 tgcccttaaa cgcctggggt aatgactctc tagcttgagg catcaaataa aacgaaaggc
     2161 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
     2221 tagggstgcg catgatctag agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     2281 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     2341 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     2401 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     2461 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     2521 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2581 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2641 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2701 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2761 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2821 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2881 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2941 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3001 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3061 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3121 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     3181 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3241 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3301 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3361 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     3421 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     3481 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     3541 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     3601 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3661 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3721 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3781 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3841 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3901 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3961 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4021 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4081 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4141 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4201 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4261 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4321 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     4381 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     4441 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     4501 aaataggcgt atcacgaggc cctttcgtct tcac
//
