LOCUS       sequence-451283-        5151 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses human KCNQ1 in Xenopus laevis oocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5151)
  TITLE     KCNQ1, an unexpected mediator of sex-dependent perception of 
            moderate cold temperatures
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5151)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pSGEM hKCNQ1
FEATURES             Location/Qualifiers
     source          1..5151
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     5'UTR           34..76
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     3'UTR           2257..2382
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     promoter        complement(2476..2494)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2477..2494)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2512..2528)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2512..2528)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2525..2547)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2536..2552
                     /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(2560..2566,2567..2584,2585..2590))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2605..2626
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2743..2760)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2914..3502)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2994..3013)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3673..4464,4465..4533))
                     /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     4296..4315
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4534..4638)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4706..4724
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4762..4784)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4884..4903
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5097..5119
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5111..5128
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5112..5128
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5135..5151,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(5135..5151,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaatta attcgagctg cgtacccagc ttgcttgttc tttttgcaga agctcagaat
       61 aaacgctcaa ctttggcaga tcaattcccc ccaccgcggt ggcggccgct ctagtgctag
      121 tggatctccc gggctgcagg aattcgatct agcaccatgg ccgcggcctc ctccccgccc
      181 agggccgaga ggaagcgctg gggttggggc cgcctgccag gcgcccggcg gggcagcgcg
      241 ggcctggcca agaagtgccc cttctcgctg gagctggcgg agggcggccc ggcgggcggc
      301 gcgctctacg cgcccatcgc gcccggcgcc ccaggtcccg cgccccctgc gtccccggcc
      361 gcgcccgccg cgcccccagt tgcctccgac cttggcccgc ggccgccggt gagcctagac
      421 ccgcgcgtct ccatctacag cacgcgccgc ccggtgttgg cgcgcaccca cgtccagggc
      481 cgcgtctaca acttcctgga gcgtcccacc ggctggaaat gcttcgttta ccacttcgcc
      541 gtcttcctca tcgtcctggt ctgcctcatc ttcagcgtgc tgtccaccat cgagcagtat
      601 gccgccctgg ccacggggac tctcttctgg atggagatcg tgctggtggt gttcttcggg
      661 acggagtacg tcgtacgcct ctggtccgcc ggctgccgca gcaagtacgt gggcctctgg
      721 gggcggctgc gctttgcccg gaagcccatt tccatcatcg atctcatcgt ggtcgtggcc
      781 tccatggtgg tcctctgcgt gggctccaag gggcaggtgt ttgccacgtc ggccatcagg
      841 ggcatccgct tcctgcagat cctgaggatg ctacacgtcg accgccaggg aggcacctgg
      901 aggctcctgg gctccgtggt cttcatccac cgccaggagc tgataaccac cctgtacatc
      961 ggcttcctgg gcctcatctt ctcgagctac tttgtgtacc tggctgagaa ggacgcggtg
     1021 aacgagtcag gccgcgtgga gttcggcagc tacgcagatg cattgtggtg gggggtggtc
     1081 acagtcacca ccatcggcta tggggacaag gtaccccaga cgtgggtcgg gaagaccatc
     1141 gcctcctgct tctctgtctt tgccatctcc ttctttgcgc tcccagcggg gattcttgga
     1201 tccgggtttg ccctgaaggt gcagcagaag cagaggcaga agcacttcaa ccggcagatc
     1261 ccggcggcag cctcactcat tcagaccgca tggaggtgct atgctgccga gaaccccgac
     1321 tcctccacct ggaagatcta catccggaag gccccccgga gccacactct gctgtcaccc
     1381 agccccaaac ccaagaagtc tgtggtggta aagaaaaaaa agttcaagct ggacaaagac
     1441 aatggggtga ctcctggaga gaagatgctc acagtccccc atatcacgtg cgacccccca
     1501 gaagagcggc ggctggacca cttctctgtc gacggctatg acagttctgt aaggaagagc
     1561 ccaacactgc tggaagtgag catgccccat ttcatgagaa ccaacagctt cgccgaggac
     1621 ctggacctgg aaggggagac tctgctgaca cccatcaccc acatctcaca gctgcgggaa
     1681 caccatcggg ccaccattaa ggtcattcga cgcatgcagt actttgtggc caagaagaaa
     1741 ttccagcaag cgcggaagcc ttacgatgtg cgggacgtca ttgagcagta ctcgcagggc
     1801 cacctcaacc tcatggtgcg catcaaggag ctgcagagga ggctggacca gtccattggg
     1861 aagccctcac tgttcatctc cgtctcagaa aagagcaagg atcgcggcag caacacgatc
     1921 ggcgcccgcc tgaaccgagt agaagacaag gtgacgcagc tggaccagag gctggcactc
     1981 atcaccgaca tgcttcacca gctgctctcc ttgcacggtg gcagcacccc cggcagcggc
     2041 ggccccccca gagagggcgg ggcccacatc acccagccct gcggcagtgg cggctccgtc
     2101 gaccctgagc tattcctgcc cagcaacacc ctgcccacct acgagcagct gaccgtgccc
     2161 aggaggggcc ccgatgaggg gtcctgagga ggatcatcaa gcttatcgat accgtcgaca
     2221 tcgagggggg gcccgagctt gatctggtta ccactaaacc agcctcaaga acacccgaat
     2281 ggagtctcta agctacataa taccaactta cactttacaa aatgttgtcc cccaaaatgt
     2341 agccattcgt atctgctcct aataaaaaga aagtttcttc acattctaaa aaaaaaaaaa
     2401 aaaaaaaaaa aaaaaccccc cccccctgca ggcatgcagt aattaaaggc catgatggcc
     2461 agctagcttg agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct
     2521 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2581 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2641 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2701 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2761 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2821 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2881 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2941 gcatcacaaa aatcgacgct caagtaagag gtggcgaaac ccgacaggac tataaagata
     3001 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3061 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     3121 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     3181 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     3241 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     3301 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3361 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3421 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3481 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3541 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3601 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3661 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3721 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3781 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3841 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3901 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3961 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4021 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4081 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4141 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4201 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4261 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4321 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4381 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4441 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4501 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4561 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4621 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     4681 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     4741 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     4801 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     4861 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     4921 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     4981 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     5041 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     5101 gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat a
//
