LOCUS       sequence-82526-e        6496 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses alpha subunit of D540K hERG1a K channel in Xenopus 
            oocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6496)
  AUTHORS   Sanguinetti MC, Xu QP
  TITLE     Mutations of the S4-S5 linker alter activation properties of HERG 
            potassium channels expressed in Xenopus oocytes.
  JOURNAL   J Physiol. 1999 Feb 1;514 ( Pt 3):667-75.
  PUBMED    9882738
REFERENCE   2  (bases 1 to 6496)
  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: "J 
            Physiol."; date: "1999-02-1"; volume: "514 ( Pt 3)"; pages: "667-75"
COMMENT     Sequence Label: pSP64-hERG1a D540K
FEATURES             Location/Qualifiers
     source          1..6496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3565..3581)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3565..3581)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3578..3600)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3589..3605
                     /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(3613..3619,3620..3637,3638..3643))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3658..3679
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3796..3813)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3967..4555)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4047..4066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4726..5517,5518..5586))
                     /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     5349..5368
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5587..5691)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5759..5777
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5815..5837)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5937..5956
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6171..6190
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(6480..6496,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(6480..6496,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaatacaagc ttctcaggat gccggtgcgg aggggccacg tcgcgccgca gaacaccttc
       61 ctggacacca tcatccgcaa gtttgagggc cagagccgta agttcatcat cgccaacgct
      121 cgggtggaga actgcgccgt catctactgc aacgacggct tctgcgagct gtgcggctac
      181 tcgcgggccg aggtgatgca gcgaccctgc acctgcgact tcctgcacgg gccgcgcacg
      241 cagcgccgcg ctgccgcgca gatcgcgcag gcactgctgg gcgccgagga gcgcaaagtg
      301 gaaatcgcct tctaccggaa agatgggagc tgcttcctat gtctggtgga tgtggtgccc
      361 gtgaagaacg aggatggggc tgtcatcatg ttcatcctca atttcgaggt ggtgatggag
      421 aaggacatgg tggggtcccc ggctcatgac accaaccacc ggggcccccc caccagctgg
      481 ctggccccag gccgcgccaa gaccttccgc ctgaagctgc ccgcgctgct ggcgctgacg
      541 gcccgggagt cgtcggtgcg gtcgggcggc gcgggcggcg cgggcgcccc gggggccgtg
      601 gtggtggacg tggacctgac gcccgcggca cccagcagcg agtcgctggc cctggacgaa
      661 gtgacagcca tggacaacca cgtggcaggg ctcgggcccg cggaggagcg gcgtgcgctg
      721 gtgggtcccg gctctccgcc ccgcagcgcg cccggccagc tcccatcgcc ccgggcgcac
      781 agcctcaacc ccgacgcctc gggctccagc tgcagcctgg cccggacgcg ctcccgagaa
      841 agctgcgcca gcgtgcgccg cgcctcgtcg gccgacgaca tcgaggccat gcgcgccggg
      901 gtgctgcccc cgccaccgcg ccacgccagc accggggcca tgcacccact gcgcagcggc
      961 ttgctcaact ccacctcgga ctccgacctc gtgcgctacc gcaccattag caagattccc
     1021 caaatcaccc tcaactttgt ggacctcaag ggcgacccct tcttggcttc gcccaccagt
     1081 gaccgtgaga tcatagcacc taagataaag gagcgaaccc acaatgtcac tgagaaggtc
     1141 acccaggtcc tgtccctggg cgccgacgtg ctgcctgagt acaagctgca ggcaccgcgc
     1201 atccaccgct ggaccatcct gcattacagc cccttcaagg ccgtgtggga ctggctcatc
     1261 ctgctgctgg tcatctacac ggctgtcttc acaccctact cggctgcctt cctgctgaag
     1321 gagacggaag aaggcccgcc tgctaccgag tgtggctacg cctgccagcc gctggctgtg
     1381 gtggacctca tcgtggacat catgttcatt gtggacatcc tcatcaactt ccgcaccacc
     1441 tacgtcaatg ccaacgagga ggtggtcagc caccccggcc gcatcgccgt ccactacttc
     1501 aagggctggt tcctcatcga catggtggcc gccatcccct tcgacctgct catcttcggc
     1561 tctggctctg aggagctgat cgggctgctg aagactgcgc ggctgctgcg gctggtgcgc
     1621 gtggcgcgga agctgaaacg ctactcagag tacggcgcgg ccgtgctgtt cttgctcatg
     1681 tgcacctttg cgctcatcgc gcactggcta gcctgcatct ggtacgccat cggcaacatg
     1741 gagcagccac acatggactc acgcatcggc tggctgcaca acctgggcga ccagataggc
     1801 aaaccctaca acagcagcgg cctgggcggc ccctccatca aggacaagta tgtgacggcg
     1861 ctctacttca ccttcagcag cctcaccagt gtgggcttcg gcaacgtctc tcccaacacc
     1921 aactcagaga agatcttctc catctgcgtc atgctcattg gctccctcat gtatgctagc
     1981 atcttcggca acgtgtcggc catcatccag cggctgtact cgggcacagc ccgctaccac
     2041 acacagatgc tgcgggtgcg ggagttcatc cgcttccacc agatccccaa tcccctgcgc
     2101 cagcgcctcg aggagtactt ccagcacgcc tggtcctaca ccaacggcat cgacatgaac
     2161 gcggtgctga agggcttccc tgagtgcctg caggctgaca tctgcctgca cctgaaccgc
     2221 tcactgctgc agcactgcaa acccttccga ggggccacca agggctgcct tcgggccctg
     2281 gccatgaagt tcaagaccac acatgcaccg ccaggggaca cactggtgca tgctggggac
     2341 ctgctcaccg ccctgtactt catctcccgg ggctccatcg agatcctgcg gggcgacgtc
     2401 gtcgtggcca tcctggggaa gaatgacatc tttggggagc ctctgaacct gtatgcaagg
     2461 cctggcaagt cgaacgggga tgtgcgggcc ctcacctact gtgacctaca caagatccat
     2521 cgggacgacc tgctggaggt gctggacatg taccctgagt tctccgacca cttctggtcc
     2581 agcctggaga tcaccttcaa cctgcgagat accaacatga tcccgggctc ccccggcagt
     2641 acggagttag agggtggctt cagtcggcaa cgcaagcgca agttgtcctt ccgcaggcgc
     2701 acggacaagg acacggagca gccaggggag gtgtcggcct tggggccggg ccgggcgggg
     2761 gcagggccga gtagccgggg ccggccgggg gggccgtggg gggagagccc gtccagtggc
     2821 ccctccagcc ctgagagcag tgaggatgag ggcccaggcc gcagctccag ccccctccgc
     2881 ctggtgccct tctccagccc caggcccccc ggagagccgc cgggtgggga gcccctgatg
     2941 gaggactgcg agaagagcag cgacacttgc aaccccctgt caggcgcctt ctcaggagtg
     3001 tccaacattt tcagcttctg gggggacagt cggggccgcc agtaccagga gctccctcga
     3061 tgccccgccc ccacccccag cctcctcaac atccccctct ccagcccggg tcggcggccc
     3121 cggggcgacg tggagagcag gctggatgcc ctccagcgcc agctcaacag gctggagacc
     3181 cggctgagtg cagacatggc cactgtcctg cagctgctac agaggcagat gacgctggtc
     3241 ccgcccgcct acagtgctgt gaccaccccg gggcctggcc ccacttccac atccccgctg
     3301 ttgcccgtca gccccctccc caccctcacc ttggactcgc tttctcaggt ttcccagttc
     3361 atggcgtgtg aggagctgcc cccgggggcc ccagagcttc cccaagaagg ccccacacga
     3421 cgcctctccc taccgggcca gctgggggcc ctcacctccc agcccctgca cagacacggc
     3481 tcggacccgg gcagttagtg gggctgccca gtgtggacac gtggctcacc cggatccccg
     3541 ggcgagctcg aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3601 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3661 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3721 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3781 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3841 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3901 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3961 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4021 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4081 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4141 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4201 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4261 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4321 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4381 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     4441 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4501 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4561 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4621 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4681 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4741 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4801 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4861 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4921 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4981 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5041 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5101 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5161 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5221 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5281 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5341 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5401 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5461 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5521 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5581 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5641 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5701 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     5761 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     5821 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     5881 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     5941 atcagagcag attgtactga gagtgcacca tatcgacgct ctcccttatg cgactcctgc
     6001 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     6061 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     6121 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     6181 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     6241 gcgtagagga tctggctagc gatgaccctg ctgattggtt cgctgaccat ttccgggtgc
     6301 gggacggcgt taccagaaac tcagaaggtt cgtccaacca aaccgactct gacggcagtt
     6361 tacgagagag atgatagggt ctgcttcagt aagccagatg ctacacaatt aggcttgtac
     6421 atattgtcgt tagaacgcgg ctacaattaa tacataacct tatgtatcat acacatacga
     6481 tttaggtgac actata
//
