LOCUS       sequence-412911-        5511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cav2.1 N-terminus (amino acids 1-100) with R57A and R59A mutations 
            that prevent dominant-negative suppression of Cav2.1 currents and a 
            C-terminal CAAX (last 10 aa of H.Ras) motif and a HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5511)
  AUTHORS   Page KM, Heblich F, Margas W, Pratt WS, Nieto-Rostro M, Chaggar K, 
            Sandhu K, Davies A, Dolphin AC
  TITLE     N terminus is key to the dominant negative suppression of Ca(V)2 
            calcium channels: implications for episodic ataxia type 2.
  JOURNAL   J Biol Chem. 2010 Jan 8;285(2):835-44. doi: 10.1074/jbc.M109.065045.
            Epub 2009 Nov 10.
  PUBMED    19903821
REFERENCE   2  (bases 1 to 5511)
  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: 
            "10.1074/jbc.M109.065045"; journalName: "J Biol Chem"; date: 
            "2010-01-8- 8"; volume: "285"; issue: "2"; pages: "835-44"
COMMENT     Sequence Label: An R57A, R59A HA CAAX pMT2
FEATURES             Location/Qualifiers
     source          1..5511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(7..336)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      21..156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     321..341
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          805..1034
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1017..1036
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1394..1420
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1566..2129
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    2316..2450
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2353..2372)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2407..2426
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3123..3140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3294..3882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3374..3393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4053..4844,4845..4913))
                     /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     4676..4695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4914..5018)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5086..5104
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5142..5164)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5264..5283
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5477..5499
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5491..5508
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5492..5508
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
       61 agaggccgag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
      121 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
      181 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      241 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
      301 ggagcctggg gactttccac accctaactg acacacattc cacaggatcc ggtcgcgcga
      361 atttcgagcg gtgttccgcg gtcctcctcg tatagaaact cggaccactc tgagacgaag
      421 gctcgcgtcc aggccagcac gaaggaggct aagtgggagg ggtagcggtc gttgtccact
      481 agggggtcca ctcgctccag ggtgtgaaga cacatgtcgc cctcttcggc atcaaggaag
      541 gtgattggtt tataggtgta ggccacgtga ccgggtgttc ctgaaggggg gctataaaag
      601 ggggtggggg cgcgttcgtc ctcactctct tccgcatcgc tgtctgcgag ggccagctgt
      661 tgggctcgcg gttgaggaca aactcttcgc ggtctttcca gtactcttgg atcggaaacc
      721 cgtcggcctc cgaacggtac tccgccaccg agggacctga gcgagtccgc atcgaccgga
      781 tcggaaaacc tctcgactgt tggggtgagt actccctctc aaaagcgggc atgacttctg
      841 cgctaagatt gtcagtttcc aaaaacgagg aggatttgat attcacctgg cccgcggtga
      901 tgcctttgag ggtggccgcg tccatctggt cagaaaagac aatctttttg ttgtcaagct
      961 tgaggtgtgg caggcttgag atctggccat acacttgagt gacaatgaca tccactttgc
     1021 ctttctctcc acaggtgtcc actcccaggt ccaactgcag gcgagcctga attgcggccg
     1081 cactagtatg gcccgctttg gagacgagat gccgggccgc tacggcgcag gcggaggagg
     1141 ctcagggccg gccgccgggg tggtcgtggg cgccgcgggc ggccgaggag ccgggggcag
     1201 ccggcagggc gggcagcccg gagcgcagag gatgtacaag cagtcgatgg cgcaggcagc
     1261 ggcgaccatg gccctctaca accccatccc tgtccgccag aactgcctca cggtcaaccg
     1321 ctccctgttc ctcttcagtg aagacaacgt ggtgagaaaa tacgccaaaa agatcacgga
     1381 atggcctcga gtctaccctt acgacgtccc tgactacgcc gttcgaggct gcatgagctg
     1441 caagtgtgtg ctctcctgaa ttccgggggg gggggggggg gggacagctc agggctgcga
     1501 tttcgcgcca aacttgacgg caatcctagc gtgaaggctg gtaggatttt atccccgctg
     1561 ccatcatggt tcgaccattg aactgcatcg tcgccgtgtc ccaaaatatg gggattggca
     1621 agaacggaga cctaccctgg cctccgctca ggaacgagtt caagtacttc caaagaatga
     1681 ccacaacctc ttcagtggaa ggtaaacaga atctggtgat tatgggtagg aaaacctggt
     1741 tctccattcc tgagaagaat cgacctttaa aggacagaat taatatagtt ctcagtagag
     1801 aactcaaaga accaccacga ggagctcatt ttcttgccaa aagtttggat gatgccttaa
     1861 gacttattga acaaccggaa ttggcaagta aagtagacat ggtttggata gtcggaggca
     1921 gttctgttta ccaggaagcc atgaatcaac caggccacct cagactcttt gtgacaagga
     1981 tcatgcagga atttgaaagt gacacgtttt tcccagaaat tgatttgggg aaatataaac
     2041 ttctcccaga atacccaggc gtcctctctg aggtccagga ggaaaaaggc atcaagtata
     2101 agtttgaagt ctacgagaag aaagactaac aggaagatgc tttcaagttc tctgctcccc
     2161 tcctaaagct atgcattttt ataagaccat gggacttttg ctggctttag atcataatca
     2221 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2281 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2341 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2401 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cccggccaac
     2461 ggtctggtga cccggctgcg agagctcggt gtacctgaga cgcgagtaag cccttgagtc
     2521 aaagacgtag tcgttgcaag tccgcaccag gtactgatat cccaccaaaa agtgcggcgg
     2581 cggctggcgg tagaggggcc agcgtagggt ggccggggct ccgggggcga ggtcttccaa
     2641 cataaggcga tgatatccgt agatgtacct ggacatccag gtgatgccgg cggcggtggt
     2701 ggaggcgcgc ggaaagtcgc ggacgcggtt ccagatgttg cgcagcggca aaaagtgctc
     2761 catggtcggg acgctctggc cggtgaggcg tgcgcagtcg ttgacgctct agaccgtgca
     2821 aaaggagagc ctgtaagcgg gcactcttcc gtggtctggt ggataaattc gcaagggtat
     2881 catggcggac gaccggggtt cgaaccccgg atccggccgt ccgccgtgat ccatgcggtt
     2941 accgcccgcg tgtcgaaccc aggtgtgcga cgtcagacaa cgggggagcg ctccttttgg
     3001 cttccttcca ggcgcggcgg ctgctgcgct agcttttttg gcgagctcga attaattctg
     3061 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3121 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3181 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3241 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3301 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3361 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3421 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3481 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3541 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3601 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3661 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3721 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3781 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3841 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3901 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3961 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4021 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4081 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4141 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4201 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4261 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4321 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4381 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4441 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4501 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4561 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4621 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4681 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4741 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4801 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4861 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4921 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4981 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5041 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     5101 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     5161 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     5221 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     5281 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     5341 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     5401 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     5461 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtgc c
//
