LOCUS       sequence-445132-        4329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription for Oocyte expression of mTRAAK(S93C/Q233K).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4329)
  AUTHORS   Deal PE, Lee H, Mondal A, Lolicato M, Mendonca PRF, Black H, Jang S,
            El-Hilali X, Bryant C, Isacoff EY, Renslo AR, Minor DL Jr
  TITLE     Development of covalent chemogenetic K(2P) channel activators.
  JOURNAL   Cell Chem Biol. 2024 Jul 18;31(7):1305-1323.e9. doi: 
            10.1016/j.chembiol.2024.06.006.
  PUBMED    39029456
REFERENCE   2  (bases 1 to 4329)
  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"; doi: 
            "10.1016/j.chembiol.2024.06"; journalName: "Cell Chem Biol"; date: 
            "2024-07-18- 18"; volume: "31"; issue: "7"; pages: "1305-1323.e9"
COMMENT     Sequence Label: pGEMHE:mTRAAK(S93C/Q233K)
FEATURES             Location/Qualifiers
     source          1..4329
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(171..962,963..1031))
                     /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     794..813
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1032..1136)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1204..1222
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1260..1282)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1382..1401
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1595..1617
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1609..1626
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1610..1626
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1633..1652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1633..1651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           1683..1725
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           3036..3161
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     CDS             3229..3255
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3303..3321)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3304..3321)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3339..3355)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3339..3355)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3352..3374)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3363..3379
                     /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(3387..3393,3394..3411,3412..3417))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3432..3453
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3570..3587)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3741..4329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     1201 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     1261 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     1321 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     1381 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     1441 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
     1501 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     1561 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     1621 gccagtgaat tgtaatacga ctcactatag ggcgaattaa ttcgagctcg gtacccagct
     1681 tgcttgttct ttttgcagaa gctcagaata aacgctcaac tttggcagat caattccccg
     1741 gggatccgaa ttctctagag caagcttatg cgcagcacca cactcctggc tctgctggca
     1801 ctggtgctgc tttacttggt atctggggct ctagtgttcc aggctctgga gcagcctcac
     1861 gagcagcagg ctcagaagaa aatggatcat ggccgagacc agtttctgag ggaccatccc
     1921 tgtgtgagcc agaagagcct ggaggatttc atcaagctcc tggttgaagc cctgggaggg
     1981 ggcgcaaacc cagaaaccag ctggaccaat agcagcaacc actcatcagc ttggaacctg
     2041 ggctgcgcct tctttttctc ggggaccatc atcactacca tcggctatgg caatatagtc
     2101 ttacacacag atgccgggcg tctcttttgt atcttctatg cactggtggg gatcccactg
     2161 ttcgggatgc tgctggcggg agtcggggac cggctgggct cctctctgcg ccggggcatc
     2221 ggccacatcg aagcaatctt cttgaagtgg catgtgccac cggggctggt gagaagtctg
     2281 tccgcagtgc tcttcctgct gatcggctgc ctgctctttg tcctcactcc taccttcgtg
     2341 ttctcctaca tggagagctg gagcaagtta gaagccatct actttgttat agtgactctc
     2401 accactgtag gctttggcga ttatgtaccc ggcgatggca ccgggcagaa ctctccagcc
     2461 tacaagccgc tggtgtggtt ctggatcttg tttggcctag cctacttcgc ctcagtgctc
     2521 accaccatcg gcaactggtt gcgagcagtg tcccgccgaa ctcgggcaga gatgggtggc
     2581 ctaacggcac aggctgctag ctggaccggc acagtgacag cgcgagtgac ccagcgaact
     2641 gggcccagcg ccccgccgcc agagaaggag caaccactcc tgccctcctc tttgccggca
     2701 ccgcctgctg ttgttgagcc agccggcagg cccggctccc ctgcacccgc agagaaggtt
     2761 gagactccgt ccccgcccac ggcctcagct ctggattacc ccagtgagaa tctggccttc
     2821 atcgacgagt cctcagacac gcagagtgag cgtggctgtg ccctgcctcg ggctcctcgg
     2881 ggtcgccgcc gacccaaccc atccaaaaag ccttccagac cccggggtcc tgggcgactc
     2941 cgagacaagg ccgtgccggt gtaggtctag actcgagtcg acggatccgc ggcccgcgca
     3001 tgcccatggt ctctagcttg atctggttac cactaaacca gcctcaagaa cacccgaatg
     3061 gagtctctaa gctacataat accaacttac actttacaaa atgttgtccc ccaaaatgta
     3121 gccattcgta tctgctccta ataaaaagaa agtttcttca cattctaaaa aaaaaaaaaa
     3181 aaaaaaaaaa aaaaaccccc cccccccccc ctgcaggcgg cgcttaagta tccgtacgac
     3241 gtcccggact acgcgtaaga attcgcggcg cctagggcgg ccggcgcggc tagcttgagt
     3301 attctatagt gtcacctaaa tagcttggcg taatcatggt catagctgtt tcctgtgtga
     3361 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3421 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3481 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3541 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3601 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3661 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3721 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3781 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagataccg ggcgtttccc
     3841 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3901 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3961 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4021 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4081 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4141 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4201 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4261 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4321 ggatctcaa
//
