LOCUS       sequence-292317-        4315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  X. laevis expression vector. It will generate the mouse TREK-1 
            channel.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4315)
  AUTHORS   Lolicato M, Arrigoni C, Mori T, Sekioka Y, Bryant C, Clark KA, Minor
            DL Jr
  TITLE     K2P2.1 (TREK-1)-activator complexes reveal a cryptic selectivity 
            filter binding site.
  JOURNAL   Nature. 2017 Jul 20;547(7663):364-368. doi: 10.1038/nature22988. 
            Epub 2017 Jul 10.
  PUBMED    28693035
REFERENCE   2  (bases 1 to 4315)
  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: 
            "10.1038/nature22988"; journalName: "Nature"; date: "2017-07-20- 
            20"; volume: "547"; issue: "7663"; pages: "364-368"
COMMENT     Sequence Label: pGEMHE:mTREK-1(K271Q)
FEATURES             Location/Qualifiers
     source          1..4315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..86
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(124..146)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     246..265
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     459..481
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     473..490
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     474..490
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     497..516
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        497..515
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           547..589
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           1932..2057
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(2153..2171)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2154..2171)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2189..2205)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2189..2205)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2202..2224)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2213..2229
                     /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(2237..2243,2244..2261,2262..2267))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2282..2303
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2420..2437)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2591..3179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2671..2690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3350..4141,4142..4210))
                     /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     3973..3992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4211..4315)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
       61 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
      121 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
      181 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
      241 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
      301 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
      361 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
      421 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
      481 gacggccagt gaattgtaat acgactcact atagggcgaa ttaattcgag ctcggtaccc
      541 agcttgcttg ttctttttgc agaagctcag aataaacgct caactttggc agatcaattc
      601 cccggggatc cgaattctct agagcaagct tatggcggcc cctgacttgc tggatcccaa
      661 gtctgctgct cagaactcca aaccgaggct ctcattctcc tcaaaaccca ccgtgcttgc
      721 ttcccgggtg gagagtgact cggccattaa tgttatgaaa tggaagacgg tctccacgat
      781 tttcctggtg gtcgtcctct acctgatcat cggagccacg gtgttcaagg cattggagca
      841 gcctcaggag atttcccaga ggaccaccat tgtgatccag aagcagacct tcatagccca
      901 gcatgcctgc gtcaactcca ccgagctgga cgaactcatc cagcaaatag tggcagcaat
      961 aaacgcagga attatcccct taggaaacag ctccaatcaa gttagtcact gggacctcgg
     1021 aagctctttc ttctttgctg gtactgttat cacaaccata ggatttggaa acatctcccc
     1081 acgaactgaa ggtggaaaaa tattctgcat catctatgcc ttgctgggaa ttcccctctt
     1141 tggctttcta ctggctgggg ttggtgatca gctaggaact atatttggaa aaggaattgc
     1201 caaagtggaa gacacattta ttaagtggaa tgttagtcag acgaagattc gtatcatctc
     1261 caccatcatc ttcatcctgt ttggctgtgt cctctttgtg gctctccctg cggtcatatt
     1321 caagcacata gaaggctgga gcgccctgga cgctatctat tttgtggtta tcactctgac
     1381 gaccattgga tttggagact acgtggcagg tggatcagac attgaatatc tggacttcta
     1441 ccagcctgtg gtgtggttct ggatcctcgt tgggctggcc tactttgcag ctgttctgag
     1501 catgattggg gactggctac gggtgatctc taagaagacg aaggaagagg tgggagagtt
     1561 cagagcgcat gccgctgagt ggacagccaa tgtcacggcc gagttcaagg aaacgaggag
     1621 gcggctgagc gtggagatct acgacaagtt ccagcgtgcc acatccgtga agcggaagct
     1681 ctccgcagag ctggcgggca accacaacca ggaactgact ccgtgtcgtc gcaccctgtc
     1741 tgtgaaccac ctgaccagcg agagggaagt cctgcctccc ttgctgaagg ctgagagcat
     1801 ctatctgaac ggtctgacac cacactgtgc tggtgaggac atagctgtca ttgagaacat
     1861 gaagtgactc gagtcgacgg atccgcggcc cgcgcatgcc catggtctct agcttgatct
     1921 ggttaccact aaaccagcct caagaacacc cgaatggagt ctctaagcta cataatacca
     1981 acttacactt tacaaaatgt tgtcccccaa aatgtagcca ttcgtatctg ctcctaataa
     2041 aaagaaagtt tcttcacatt ctaaaaaaaa aaaaaaaaaa aaaaaaaaaa aacccggact
     2101 acgcgtaaga attcgcggcg cctagggcgg ccggcgcggc tagcttgagt attctatagt
     2161 gtcacctaaa tagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2221 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2281 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2341 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2401 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2461 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2521 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2581 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2641 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2701 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2761 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2821 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     2881 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     2941 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3001 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3061 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3121 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3181 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3241 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3301 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3361 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3421 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3481 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3541 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3601 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3661 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3721 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3781 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3841 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     3901 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     3961 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4021 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4081 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4141 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4201 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4261 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcg
//
