LOCUS       sequence-445099-        3981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro transcription for Oocyte expression of hTRAAK(S118C/Q258K).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3981)
  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 3981)
  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:hTRAAK(S118C/Q258K)
FEATURES             Location/Qualifiers
     source          1..3981
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     103..122
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     316..338
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     330..347
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     331..347
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     354..373
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        354..372
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           404..446
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     primer_bind     complement(1943..1960)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2114..2702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2194..2213)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2873..3664,3665..3733))
                     /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     3496..3515
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3734..3838)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3906..3924
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(3962..3981,1..3))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
       61 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
      121 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
      181 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
      241 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
      301 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg
      361 actcactata gggcgaatta attcgagctc ggtacccagc ttgcttgttc tttttgcaga
      421 agctcagaat aaacgctcaa ctttggcaga tcaattcccc ggggatccga attctctaga
      481 gcaagcttat gactaccgct ccacaagaac cacccgctcg cccactccaa gctggctccg
      541 gtgctggtcc tgctcctgga cgtgctatga gatcaacaac gctcctcgct ctgctggccc
      601 tcgtattgct gtacctcgtg tctggtgctc tggtattccg tgctctcgaa caaccacatg
      661 aacaacaggc tcaacgtgaa ctgggagaag tccgtgaaaa attcctccgt gcccatccgt
      721 gtgtttccga tcaagagttg ggattgctca tcaaggaagt cgccgacgct ttgggaggcg
      781 gtgccgaccc tgagacaaac tccacctcaa actcctctca ctctgcctgg gatttgggat
      841 gtgccttctt cttctctgga accatcatca ccactatcgg ttacggtaac gtggcactcc
      901 gtaccgacgc cggtagactg ttctgtattt tctacgccct cgttggtatc cctctcttcg
      961 gaatcttgtt ggctggagtc ggagatcgcc tcggttcatc attgaggcat ggcatcggtc
     1021 atattgaggc cattttcttg aaatggcacg tgccacctga attggttcgt gtactgtccg
     1081 ctatgctgtt cctcctcatc ggttgcttgc tcttcgttct cacccccact ttcgtattct
     1141 gctacatgga ggactggtct aaactggagg ccatctactt cgtgatcgtg accctcacta
     1201 ctgttggatt cggagactac gttgctggag ccgatcctag acaggattct cctgcctaca
     1261 aacctttggt gtggttctgg attctgttgg gactggcata cttcgctagt gtgctcacca
     1321 ccattggcaa ctggttgaga gtggtatcac gtcgtactcg tgctgagatg ggaggactca
     1381 ctgctcaagc tgcttcatgg accggaactg tgactgctag agtcacccaa cgtgctggac
     1441 cggctgctcc accacctgaa aaagaacaac ccctgctgcc tcctccgcct tgtcccgccc
     1501 aaccgctggg acgtcctcgt tctccttcac ctcccgaaaa ggcacaacca ccttcacccc
     1561 ctactgcttc cgctttggac tacccttctg aaaacctggc tttcatcgac gaatcttcgg
     1621 acactcaatc cgagcgtgga tgtccattgc ctagggctcc taggggccgt cgtagaccca
     1681 accctccccg taaacctgtt aggcctcgtg gacctggtag accgagagac aaaggagtcc
     1741 ctgtttgact cgagtcgacg gatccgcggc ccgcgcatgc ccatggtctc tagcttgatc
     1801 tggttaccac taaaccagcc tcaagaacac ccgaatggag tctctaagct acataatacc
     1861 aactcctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1921 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1981 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2041 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     2101 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     2161 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     2221 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     2281 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     2341 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2401 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2461 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2521 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2581 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2641 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2701 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2761 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2821 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2881 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2941 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     3001 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     3061 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     3121 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     3181 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3241 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3301 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3361 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3421 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3481 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3541 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3601 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3661 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3721 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3781 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3841 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     3901 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     3961 acctctgaca catgcagctc c
//
