LOCUS       sequence-324311-        4740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of inactivated control version of genetically 
            encoded biosensor for (pseudo)hypohalous acids and their 
            derivatives.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4740)
  AUTHORS   Kostyuk AI, Tossounian MA, Panova AS, Thauvin M, Raevskii RI, 
            Ezerina D, Wahni K, Van Molle I, Sergeeva AD, Vertommen D, 
            Gorokhovatsky AY, Baranov MS, Vriz S, Messens J, Bilan DS, Belousov 
            VV
  TITLE     Hypocrates is a genetically encoded fluorescent biosensor for 
            (pseudo)hypohalous acids and their derivatives.
  JOURNAL   Nat Commun. 2022 Jan 10;13(1):171. doi: 10.1038/s41467-021-27796-2.
  PUBMED    35013284
REFERENCE   2  (bases 1 to 4740)
  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: "2022-01-10"; volume: "13(1):171. doi"; pages: " 
            10.1038/s41467-021-27796-2"
COMMENT     Sequence Label: pQE30-HypocratesCS
FEATURES             Location/Qualifiers
     source          1..4740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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)."
     RBS             65..76
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             95..112
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     652..673
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(782..803)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1043..1062)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     terminator      1487..1581
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1625..2284
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1691..1710)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2349..2435
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2482..2504)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2590..2730
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2645..2664
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2745..2762)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2916..3504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2996..3015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3675..4466,4467..4535))
                     /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     4298..4317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4536..4640)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4708..4726
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaatttc acacagaatt
       61 cattaaagag gagaaattaa ctatgagagg atcgcatcac catcaccatc acggatccat
      121 gaacaaacac accgaacatg atactcgcga acatctcctg gcgacgggcg agcaacttag
      181 cctgcaacgt ggattcaccg ggatggggct aagcgaatta ctaaaaaccg ctgaagtgcc
      241 gaaagggtcc ttctatcact actttcgctc taaagaagcg tttggcgttg ccatgcttga
      301 gcgtcattac gccgcatatc accagcgact gactgagttg ctgcaatccg gcgaaggtaa
      361 ctaccgcgac cgcatactgg cttattacca gcaaacactg aaccagttta gccaacatgg
      421 aacctccgcc ggctacaaca gcgacaacgt ctatatcatg gccgacaagc agaagaacgg
      481 catcaaggcc aacttcaaga tccgccacaa cgtcgaggac ggcagcgtgc agctcgccga
      541 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
      601 cctgagcttc cagtccgtcc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
      661 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaacgtgga
      721 tggcggtagc ggtggcaccg gcagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      781 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      841 gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg gcaagctgcc
      901 cgtgccctgg cccaccctcg tgaccaccct cggctacggc ctgaagtgct tcgcccgcta
      961 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1021 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1081 cgagggcgac accctggtga accgcatcga gctgaagggc atcggcttca aggaggacgg
     1141 caacatcctg gggcacaagc tggagtacaa cggcaccggt agcctgacag taaaactctc
     1201 tgccgaagtg agcgatctgt cagaagatat gcgcagcgcg atggataaag gtgctcgcgg
     1261 cgtgatcgcc ctgctctctc aggcgctgga aaatggccgt gagaaccata gcttaacctt
     1321 tagtggcgaa ccgctgcaac aggcacaagt gctttacgca ctgtggcttg gcgcgaatct
     1381 gcaggccaaa atttcgcgca gtttcgagcc actggaaaac gcgctggccc atgtaaaaaa
     1441 cattattgcg acgcctgccg tttagaagct taattagctg agcttggact cctgttgata
     1501 gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg
     1561 gcgtttttta ttggtgagaa tccaagctag cttggcgaga ttttcaggag ctaaggaagc
     1621 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     1681 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     1741 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     1801 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga
     1861 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     1921 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     1981 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     2041 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     2101 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     2161 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     2221 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     2281 gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggggtaatga ctctctagct
     2341 tgaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     2401 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc ctctagagct gcctcgcgcg
     2461 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     2521 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     2581 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     2641 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac
     2701 agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg
     2761 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2821 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2881 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2941 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3001 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3061 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3121 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3181 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     3241 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     3301 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     3361 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     3421 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     3481 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     3541 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     3601 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     3661 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3721 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3781 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3841 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3901 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3961 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     4021 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     4081 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     4141 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     4201 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     4261 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     4321 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     4381 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     4441 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     4501 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     4561 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     4621 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     4681 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac
//
