LOCUS       sequence-260987-        4391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  X. laevis expression vector. It will generate the human TRAAK 
            channel.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4391)
  AUTHORS   Lolicato M, Riegelhaupt PM, Arrigoni C, Clark KA, Minor DL Jr
  TITLE     Transmembrane helix straightening and buckling underlies activation 
            of mechanosensitive and thermosensitive K(2P) channels.
  JOURNAL   Neuron. 2014 Dec 17;84(6):1198-212. doi: 
            10.1016/j.neuron.2014.11.017. Epub 2014 Dec 11.
  PUBMED    25500157
REFERENCE   2  (bases 1 to 4391)
  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.neuron.2014.11.017"; journalName: "Neuron"; date: 
            "2014-12-17- 17"; volume: "84"; issue: "6"; pages: "1198-212"
COMMENT     Sequence Label: pGEMHE:hTRAAK(Q258K)
FEATURES             Location/Qualifiers
     source          1..4391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     3'UTR           1367..1492
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     CDS             1566..1592
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1640..1658)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1641..1658)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1676..1692)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1676..1692)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1689..1711)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1700..1716
                     /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(1724..1730,1731..1748,1749..1754))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1769..1790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1907..1924)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2078..2666)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2158..2177)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2837..3628,3629..3697))
                     /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     3460..3479
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3698..3802)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3870..3888
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3926..3948)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4048..4067
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4261..4283
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4275..4292
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4276..4292
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4299..4318
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4299..4317
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           4349..4391
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
ORIGIN
        1 cagatcaatt ccccggggat ccgaattctc tagagcaagc ttatgactac cgctccacaa
       61 gaaccacccg ctcgcccact ccaagctggc tccggtgctg gtcctgctcc tggacgtgct
      121 atgagatcaa caacgctcct cgctctgctg gccctcgtat tgctgtacct cgtgtctggt
      181 gctctggtat tccgtgctct cgaacaacca catgaacaac aggctcaacg tgaactggga
      241 gaagtccgtg aaaaattcct ccgtgcccat ccgtgtgttt ccgatcaaga gttgggattg
      301 ctcatcaagg aagtcgccga cgctttggga ggcggtgccg accctgagac aaactccacc
      361 tcaaactcct ctcactctgc ctgggatttg ggatctgcct tcttcttctc tggaaccatc
      421 atcaccacta tcggttacgg taacgtggca ctccgtaccg acgccggtag actgttctgt
      481 attttctacg ccctcgttgg tatccctctc ttcggaatct tgttggctgg agtcggagat
      541 cgcctcggtt catcattgag gcatggcatc ggtcatattg aggccatttt cttgaaatgg
      601 cacgtgccac ctgaattggt tcgtgtactg tccgctatgc tgttcctcct catcggttgc
      661 ttgctcttcg ttctcacccc cactttcgta ttctgctaca tggaggactg gtctaaactg
      721 gaggccatct acttcgtgat cgtgaccctc actactgttg gattcggaga ctacgttgct
      781 ggagccgatc ctagacagga ttctcctgcc tacaaacctt tggtgtggtt ctggattctg
      841 ttgggactgg catacttcgc tagtgtgctc accaccattg gcaactggtt gagagtggta
      901 tcacgtcgta ctcgtgctga gatgggagga ctcactgctc aagctgcttc atggaccgga
      961 actgtgactg ctagagtcac ccaacgtgct ggaccggctg ctccaccacc tgaaaaagaa
     1021 caacccctgc tgcctcctcc gccttgtccc gcccaaccgc tgggacgtcc tcgttctcct
     1081 tcacctcccg aaaaggcaca accaccttca ccccctactg cttccgcttt ggactaccct
     1141 tctgaaaacc tggctttcat cgacgaatct tcggacactc aatccgagcg tggatgtcca
     1201 ttgcctaggg ctcctagggg ccgtcgtaga cccaaccctc cccgtaaacc tgttaggcct
     1261 cgtggacctg gtagaccgag agacaaagga gtccctgttt gactcgagtc gacggatccg
     1321 cggcccgcgc atgcccatgg tctctagctt gatctggtta ccactaaacc agcctcaaga
     1381 acacccgaat ggagtctcta agctacataa taccaactta cactttacaa aatgttgtcc
     1441 cccaaaatgt agccattcgt atctgctcct aataaaaaga aagtttcttc acattctaaa
     1501 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa amcccccccc cccccccctg caggcggcgc
     1561 ttaagtatcc gtacgacgtc ccggactacg cgtaagaatt cgcggcgcct agggcggccg
     1621 gcgcggctag cttgagtatt ctatagtgtc acctaaatag cttggcgtaa tcatggtcat
     1681 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     1741 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     1801 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     1861 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     1921 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     1981 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2041 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2101 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2161 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2221 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2281 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2341 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2401 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2461 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2521 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2581 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2641 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2701 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2761 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     2821 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     2881 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     2941 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3001 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3061 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3121 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3181 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3241 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3301 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3361 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3421 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3481 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3541 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3601 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3661 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3721 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3781 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa
     3841 ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg
     3901 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     3961 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     4021 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     4081 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt
     4141 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     4201 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     4261 cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg
     4321 aattaattcg agctcggtac ccagcttgct tgttcttttt gcagaagctc agaataaacg
     4381 ctcaactttg g
//
