LOCUS       sequence-82532-e        3431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses beta subunit for IKs channel in Xenopus oocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3431)
  AUTHORS   Sanguinetti MC, Curran ME, Zou A, Shen J, Spector PS, Atkinson DL, 
            Keating MT
  TITLE     Coassembly of K(V)LQT1 and minK (IsK) proteins to form cardiac I(Ks)
            potassium channel.
  JOURNAL   Nature. 1996 Nov 7;384(6604):80-3.
  PUBMED    8900283
REFERENCE   2  (bases 1 to 3431)
  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"; journalName: "Nature."; 
            date: "1996-11-7"; volume: "384(6604)"; pages: "80-3"
COMMENT     Sequence Label: pSP64-hKCNE1
FEATURES             Location/Qualifiers
     source          1..3431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..24
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6..23
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(521..537)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(521..537)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(534..556)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    545..561
                     /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(569..575,576..593,594..599))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    614..635
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(752..769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(923..1511)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1003..1022)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1682..2473,2474..2542))
                     /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     2305..2324
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2543..2647)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2715..2733
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2771..2793)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2893..2912
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3127..3146
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atacgattta ggtgacacta tagaatacaa gcttaatgcc caggatgatc ctgtctaaca
       61 ccacagcggt gacgcccttt ctgaccaagc tgtggcagga gacagttcag cagggtggca
      121 acatgtcggg cctggcccgc aggtcccccc gcagcggtga cggcaagctg gaggccctct
      181 acgtcctcat ggtactggga ttcttcggct tcttcaccct gggcatcatg ctgagctaca
      241 tccgctccaa gaagctggag cactcgaacg acccattcaa cgtctacatc gagtccgatg
      301 cctggcaaga gaaggacaag gcctatgtcc aggcccgggt cctggagagc tacaggtcgt
      361 gctatgtcgt tgaaaaccat ctggccatag aacaacccaa cacacacctt cctgagacga
      421 agccttcccc atgaacccca ccactggcta aactggatcc ccgggcgagc tcccaaaaaa
      481 aaaaaaaaaa aaaaaaaaaa aaaaccgaat tcgtaatcat gtcatagctg tttcctgtgt
      541 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
      601 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
      661 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
      721 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
      781 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
      841 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
      901 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
      961 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1021 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1081 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1141 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1201 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1261 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1321 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     1381 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     1441 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     1501 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     1561 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     1621 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     1681 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     1741 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     1801 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     1861 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     1921 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     1981 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2041 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2101 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     2161 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     2221 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2281 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     2341 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     2401 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     2461 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     2521 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     2581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     2641 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     2701 cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg
     2761 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg
     2821 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct
     2881 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatc gacgctctcc
     2941 cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt tgagcaccgc
     3001 cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg ccacggggcc
     3061 tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag cccgatcttc
     3121 cccatcggtg atgtcggcga tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc
     3181 ggccacgatg cgtccggcgt agaggatctg gctagcgatg accctgctga ttggttcgct
     3241 gaccatttcc ggggtgcgga acggcgttac cagaaactca gaaggttcgt ccaaccaaac
     3301 cgactctgac ggcagtttac gagagagatg atagggtctg cttcagtaag ccagatgcta
     3361 cacaattagg cttgtacata ttgtcgttag aacgcggcta caattaatac ataaccttat
     3421 gtatcataca c
//
