LOCUS       sequence-415935-        5475 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.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5475)
  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 5475)
  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 CAAX pMT2
FEATURES             Location/Qualifiers
     source          1..5475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(2..331)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      16..151
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     316..336
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          800..1029
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1012..1031
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1525..2088
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    2275..2409
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2312..2331)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2366..2385
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3082..3099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3253..3841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3333..3352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4012..4803,4804..4872))
                     /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     4635..4654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4873..4977)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5045..5063
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5101..5123)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5223..5242
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5436..5458
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5450..5467
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5451..5467
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
       61 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
      121 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
      181 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
      241 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
      301 ctggggactt tccacaccct aactgacaca cattccacag gatccggtcg cgcgaatttc
      361 gagcggtgtt ccgcggtcct cctcgtatag aaactcggac cactctgaga cgaaggctcg
      421 cgtccaggcc agcacgaagg aggctaagtg ggaggggtag cggtcgttgt ccactagggg
      481 gtccactcgc tccagggtgt gaagacacat gtcgccctct tcggcatcaa ggaaggtgat
      541 tggtttatag gtgtaggcca cgtgaccggg tgttcctgaa ggggggctat aaaagggggt
      601 gggggcgcgt tcgtcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc
      661 tcgcggttga ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg
      721 gcctccgaac ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga
      781 aaacctctcg actgttgggg tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta
      841 agattgtcag tttccaaaaa cgaggaggat ttgatattca cctggcccgc ggtgatgcct
      901 ttgagggtgg ccgcgtccat ctggtcagaa aagacaatct ttttgttgtc aagcttgagg
      961 tgtggcaggc ttgagatctg gccatacact tgagtgacaa tgacatccac tttgcctttc
     1021 tctccacagg tgtccactcc caggtccaac tgcaggcgag cctgaattgc ggccgcacta
     1081 gtatggcccg ctttggagac gagatgccgg gccgctacgg cgcaggcgga ggaggctcag
     1141 ggccggccgc cggggtggtc gtgggcgccg cgggcggccg aggagccggg ggcagccggc
     1201 agggcgggca gcccggagcg cagaggatgt acaagcagtc gatggcgcag gcagcggcga
     1261 ccatggccct ctacaacccc atccctgtcc gccagaactg cctcacggtc aaccgctccc
     1321 tgttcctctt cagtgaagac aacgtggtga gaaaatacgc caaaaagatc acggaatggc
     1381 ctcgaggctg catgagctgc aagtgtgtgc tctcctgaat tccggggggg gggggggggr
     1441 ksmcagctca gggctgcgat ttcgcgccaa acttgacggc aatcctagcg tgaaggctgg
     1501 taggatttta tccccgctgc catcatggtt cgaccattga actgcatcgt cgccgtgtcc
     1561 caaaatatgg ggattggcaa gaacggagac ctaccctggc ctccgctcag gaacgagttc
     1621 aagtacttcc aaagaatgac cacaacctct tcagtggaag gtaaacagaa tctggtgatt
     1681 atgggtagga aaacctggtt ctccattcct gagaagaatc gacctttaaa ggacagaatt
     1741 aatatagttc tcagtagaga actcaaagaa ccaccacgag gagctcattt tcttgccaaa
     1801 agtttggatg atgccttaag acttattgaa caaccggaat tggcaagtaa agtagacatg
     1861 gtttggatag tcggaggcag ttctgtttac caggaagcca tgaatcaacc aggccacctc
     1921 agactctttg tgacaaggat catgcaggaa tttgaaagtg acacgttttt cccagaaatt
     1981 gatttgggga aatataaact tctcccagaa tacccaggcg tcctctctga ggtccaggag
     2041 gaaaaaggca tcaagtataa gtttgaagtc tacgagaaga aagactaaca ggaagatgct
     2101 ttcaagttct ctgctcccct cctaaagcta tgcattttta taagaccatg ggacttttgc
     2161 tggctttaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     2221 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     2281 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     2341 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     2401 gtctggatcc ccggccaacg gtctggtgac ccggctgcga gagctcggtg tacctgagac
     2461 gcgagtaagc ccttgagtca aagacgtagt cgttgcaagt ccgcaccagg tactgatatc
     2521 ccaccaaaaa gtgcggcggc ggctggcggt agaggggcca gcgtagggtg gccggggctc
     2581 cgggggcgag gtcttccaac ataaggcgat gatatccgta gatgtacctg gacatccagg
     2641 tgatgccggc ggcggtggtg gaggcgcgcg gaaagtcgcg gacgcggttc cagatgttgc
     2701 gcagcggcaa aaagtgctcc atggtcggga cgctctggcc ggtgaggcgt gcgcagtcgt
     2761 tgacgctcta gaccgtgcaa aaggagagcc tgtaagcggg cactcttccg tggtctggtg
     2821 gataaattcg caagggtatc atggcggacg accggggttc gaaccccgga tccggccgtc
     2881 cgccgtgatc catgcggtta ccgcccgcgt gtcgaaccca ggtgtgcgac gtcagacaac
     2941 gggggagcgc tccttttggc ttccttccag gcgcggcggc tgctgcgcta gcttttttgg
     3001 cgagctcgaa ttaattctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3061 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3121 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3181 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3241 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3301 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3361 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3421 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3481 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3541 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3601 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3661 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3721 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3781 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3841 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3901 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3961 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4021 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4081 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4141 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4201 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4261 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4321 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4381 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4441 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4501 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4561 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4621 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4681 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4741 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4801 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4861 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4921 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4981 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     5041 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     5101 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     5161 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     5221 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
     5281 atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg
     5341 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     5401 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     5461 ggccagtgcc aagct
//
