LOCUS       sequence-445023-        4362 bp    DNA     circular SYN 15-APR-2025
DEFINITION  In vitro transcription for Oocyte expression of mTREK-1 
            (S131A/C159A/C219A/C365A/C399A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4362)
  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 4362)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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:mTREK-1 (S131A/C159A/C219A/C365A/C399A)
FEATURES             Location/Qualifiers
     source          1..4362
                     /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           3068..3193
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     CDS             3262..3288
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3336..3354)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3337..3354)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3372..3388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3372..3388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3385..3407)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3396..3412
                     /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(3420..3426,3427..3444,3445..3450))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3465..3486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3603..3620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3774..4362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3854..3873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 gcggcccctg acttgctgga tcccaagtct
     1801 gctgctcaga actccaaacc gaggctctca ttctcctcaa aacccaccgt gcttgcttcc
     1861 cgggtggaga gtgactcggc cattaatgtt atgaaatgga agacggtctc cacgattttc
     1921 ctggtggtcg tcctctacct gatcatcgga gccacggtgt tcaaggcatt ggagcagcct
     1981 caggagattt cccagaggac caccattgtg atccagaagc agaccttcat agcccagcat
     2041 gcctgcgtca actccaccga gctggacgaa ctcatccagc aaatagtggc agcaataaac
     2101 gcaggaatta tccccttagg aaacagctcc aatcaagtta gtcactggga cctcggagcc
     2161 tctttcttct ttgctggtac tgttatcaca accataggat ttggaaacat ctccccacga
     2221 actgaaggtg gaaaaatatt cgccatcatc tatgccttgc tgggaattcc cctctttggc
     2281 tttctactgg ctggggttgg tgatcagcta ggaactatat ttggaaaagg aattgccaaa
     2341 gtggaagaca catttattaa gtggaatgtt agtcagacga agattcgtat catctccacc
     2401 atcatcttca tcctgtttgg cgctgtcctc tttgtggctc tccctgcggt catattcaag
     2461 cacatagaag gctggagcgc cctggacgct atctattttg tggttatcac tctgacgacc
     2521 attggatttg gagactacgt ggcaggtgga tcagacattg aatatctgga cttctacaag
     2581 cctgtggtgt ggttctggat cctcgttggg ctggcctact ttgcagctgt tctgagcatg
     2641 attggggact ggctacgggt gatctctaag aagacgaagg aagaggtggg agagttcaga
     2701 gcgcatgccg ctgagtggac agccaatgtc acggccgagt tcaaggaaac gaggaggcgg
     2761 ctgagcgtgg agatctacga caagttccag cgtgccacat ccgtgaagcg gaagctctcc
     2821 gcagagctgg cgggcaacca caaccaggaa ctgactccgg ccaggaggac cctgtctgtg
     2881 aaccacctga ccagcgagag ggaagtcctg cctcccttgc tgaaggctga gagcatctat
     2941 ctgaacggtc tgacaccaca cgccgctggt gaggacatag ctgtcattga gaacatgaag
     3001 tgactcgagt cgacggatcc gcggcccgcg catgcccatg gtctctagct tgatctggtt
     3061 accactaaac cagcctcaag aacacccgaa tggagtctct aagctacata ataccaactt
     3121 acactttaca aaatgttgtc ccccaaaatg tagccattcg tatctgctcc taataaaaar
     3181 aaagtttctt cacattctaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaan nncccccccc
     3241 cccctgcagg cggcgcttaa gtatccgtac gacgtcccgg actacgcgta agaattcgcg
     3301 gcgcctaggg cggccggcgc ggctagcttg agtattctat agtgtcacct aaatagcttg
     3361 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3421 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3481 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3541 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3601 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3661 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3721 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3781 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3841 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3901 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3961 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4021 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4081 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4141 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4201 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4261 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4321 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aa
//
