LOCUS       sequence-324303-        5422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of non-targeted 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 5422)
  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 5422)
  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: pCS2+-HypocratesCS
FEATURES             Location/Qualifiers
     source          1..5422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     631..652
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(761..782)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1022..1041)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    1474..1608
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1498..1517)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1552..1571
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1721..1741)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1721..1739)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1760..1776)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1760..1776)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1773..1795)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1784..1800
                     /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)."
     promoter        complement(join(1808..1814,1815..1832,1833..1838))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1853..1874
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1991..2008)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2162..2750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2242..2261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2921..3712,3713..3781))
                     /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     3544..3563
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3782..3886)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3912..4367)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4049..4070)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4261..4280
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4438..5422
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga tgccaccatg aacaaacaca ccgaacatga
      121 tactcgcgaa catctcctgg cgacgggcga gcaacttagc ctgcaacgtg gattcaccgg
      181 gatggggcta agcgaattac taaaaaccgc tgaagtgccg aaagggtcct tctatcacta
      241 ctttcgctct aaagaagcgt ttggcgttgc catgcttgag cgtcattacg ccgcatatca
      301 ccagcgactg actgagttgc tgcaatccgg cgaaggtaac taccgcgacc gcatactggc
      361 ttattaccag caaacactga accagtttag ccaacatgga acctccgccg gctacaacag
      421 cgacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggcca acttcaagat
      481 ccgccacaac gtcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
      541 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcttcc agtccgtcct
      601 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
      661 cgggatcact ctcggcatgg acgagctgta caacgtggat ggcggtagcg gtggcaccgg
      721 cagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
      781 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
      841 gctgaccctg aagctgatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
      901 gaccaccctc ggctacggcc tgaagtgctt cgcccgctac cccgaccaca tgaagcagca
      961 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1021 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1081 ccgcatcgag ctgaagggca tcggcttcaa ggaggacggc aacatcctgg ggcacaagct
     1141 ggagtacaac ggcaccggta gcctgacagt aaaactctct gccgaagtga gcgatctgtc
     1201 agaagatatg cgcagcgcga tggataaagg tgctcgcggc gtgatcgccc tgctctctca
     1261 ggcgctggaa aatggccgtg agaaccatag cttaaccttt agtggcgaac cgctgcaaca
     1321 ggcacaagtg ctttacgcac tgtggcttgg cgcgaatctg caggccaaaa tttcgcgcag
     1381 tttcgagcca ctggaaaacg cgctggccca tgtaaaaaac attattgcga cgcctgccgt
     1441 ttagtctaga actatagtga gtcgtattac gtagatccag acatgataag atacattgat
     1501 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1561 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     1621 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta attcgcggcc
     1681 gcggcgccaa tgcattgggc ccggtaccca gcttttgttc cctttagtga gggttaattg
     1741 cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1801 ttccacacaa catacgagcc gggagcataa agtgtaaagc ctggggtgcc taatgagtga
     1861 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1921 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1981 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2041 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2101 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2161 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2221 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2281 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2341 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2401 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2461 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2521 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2581 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2641 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2701 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2761 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2821 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2881 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2941 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3001 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3061 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3121 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3181 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3241 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3301 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3361 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3421 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3481 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3541 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3601 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3661 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3721 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3781 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3841 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3901 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3961 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     4021 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     4081 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     4141 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     4201 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4261 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4321 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     4381 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc
     4441 gaccgccaat tcaatatggc gtatatggac tcatgccaat tcaatatggt ggatctggac
     4501 ctgtgccaat tcaatatggc gtatatggac tcgtgccaat tcaatatggt ggatctggac
     4561 cccagccaat tcaatatggc ggacttggca ccatgccaat tcaatatggc ggacctggca
     4621 ctgtgccaac tggggagggg tctacttggc acggtgccaa gtttgaggag gggtcttggc
     4681 cctgtgccaa gtccgccata ttgaattggc atggtgccaa taatggcggc catattggct
     4741 atatgccagg atcaatatat aggcaatatc caatatggcc ctatgccaat atggctattg
     4801 gccaggttca atactatgta ttggccctat gccatatagt attccatata tgggttttcc
     4861 tattgacgta gatagcccct cccaatgggc ggtcccatat accatatatg gggcttccta
     4921 ataccgccca tagccactcc cccattgacg tcaatggtct ctatatatgg tctttcctat
     4981 tgacgtcata tgggcggtcc tattgacgta tatggcgcct cccccattga cgtcaattac
     5041 ggtaaatggc ccgcctggct caatgcccat tgacgtcaat aggaccaccc accattgacg
     5101 tcaatgggat ggctcattgc ccattcatat ccgttctcac gccccctatt gacgtcaatg
     5161 acggtaaatg gcccacttgg cagtacatca atatctatta atagtaactt ggcaagtaca
     5221 ttactattgg aagtacgcca gggtacattg gcagtactcc cattgacgtc aatggcggta
     5281 aatggcccgc gatggctgcc aagtacatcc ccattgacgt caatggggag gggcaatgac
     5341 gcaaatgggc gttccattga cgtaaatggg cggtaggcgt gcctaatggg aggtctatat
     5401 aagcaatgct cgtttaggga ac
//
