LOCUS       sequence-292311-        3992 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  X. laevis expression vector. It will generate the TREK-1 channel.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3992)
  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 3992)
  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_cryst
FEATURES             Location/Qualifiers
     source          1..3992
                     /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"
     CDS             1756..1782
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1830..1848)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1831..1848)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1866..1882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1866..1882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1879..1901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1890..1906
                     /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(1914..1920,1921..1938,1939..1944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1959..1980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2097..2114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2268..2856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2348..2367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3027..3818,3819..3887))
                     /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     3650..3669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3888..3992)
                     /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 tatgtcattc tcctcaaaac ccaccgtgct
      661 tgcttcccgg gtggagagtg actcggccat taatgttatg aaatggaaga cggtctccac
      721 gattttcctg gtggtcgtcc tctacctgat catcggagcc acggtgttca aggcattgga
      781 gcagcctcag gagatttccc agaggaccac cattgtgatc cagagggaga agttcttacg
      841 cgcgcatccc tgcgtcagcg accaggagct ggacgaactc atccagcaaa tagtggcagc
      901 aataaacgca ggaattatcc ccttaggagc cagctccaat caagttagtc actgggacct
      961 cggaagctct ttcttctttg ctggtactgt tatcacaacc ataggatttg gaaacatctc
     1021 cccacgaact gaaggtggaa aaatattctg catcatctat gccttgctgg gaattcccct
     1081 ctttggcttt ctactggctg gggttggtga tcagctagga actatatttg gaaaaggaat
     1141 tgccaaagtg gaagacacat ttattaagtg gaatgttagt cagacgaaga ttcgtatcat
     1201 ctccaccatc atcttcatcc tgtttggctg tgtcctcttt gtggctctcc ctgcggtcat
     1261 attcaagcac atagaaggct ggagcgccct ggacgctatc tattttgtgg ttatcactct
     1321 gacgaccatt ggatttggag actacgtggc aggtggatca gacattgaat atctggactt
     1381 ctacaagcct gtggtgtggt tctggatcct cgttgggctg gcctactttg cagctgttct
     1441 gagcatgatt ggggactggc tacgggtgat cgctaagaag acgaaggaag cggtgggaga
     1501 gttcagagcg catgccgctg agtggacagc caatgtcact taatgactcg agtcgacgga
     1561 tccgcggccc gcgcatgccc atggtctcta gcttgatctg gttaccacta aaccagcctc
     1621 aagaacaccc gaatggagtc tctaagctac ataataccaa cttacacttt acaaaatgtt
     1681 gtcccccaaa atgtagccat tcgtatctgc tcctaataaa aagaaagtcc cccccccctg
     1741 caggcggcgc ttaagtatcc gtacgacgtc ccggactacg cgtaagaatt cgcggcgcct
     1801 agggcggccg gcgcggctag cttgagtatt ctatagtgtc acctaaatag cttggcgtaa
     1861 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1921 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1981 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     2041 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2101 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2161 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2221 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2281 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2341 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2401 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2461 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2521 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2581 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2641 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2701 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2761 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2821 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2881 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2941 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3001 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3061 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3121 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3181 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3241 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3301 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3361 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3421 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3481 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3541 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3601 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3661 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3721 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3781 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3841 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3901 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3961 atttagaaaa ataaacaaat aggggttccg cg
//
