LOCUS       sequence-415913-        5645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cav2.2 N-terminus (amino acids 1-95) with R52A and R54A mutations 
            that prevent dominant-negative suppression of Cav2.2 currents by 
            truncated Cav2.2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5645)
  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 5645)
  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: Bn R52A, R54A pMT2
FEATURES             Location/Qualifiers
     source          1..5645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             91..654
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    841..975
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(878..897)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     932..951
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1648..1665)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1819..2407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1899..1918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2578..3369,3370..3438))
                     /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     3201..3220
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3439..3543)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3611..3629
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3667..3689)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3789..3808
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4002..4024
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4016..4033
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4017..4033
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4043..4372)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4057..4192
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4357..4377
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          4841..5070
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     5053..5072
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 ggggggggac agctcagggc tgcgatttcg cgccaaactt gacggcaatc ctagcgtgaa
       61 ggctggtagg attttatccc cgctgccatc atggttcgac cattgaactg catcgtcgcc
      121 gtgtcccaaa atatggggat tggcaagaac ggagacctac cctggcctcc gctcaggaac
      181 gagttcaagt acttccaaag aatgaccaca acctcttcag tggaaggtaa acagaatctg
      241 gtgattatgg gtaggaaaac ctggttctcc attcctgaga agaatcgacc tttaaaggac
      301 agaattaata tagttctcag tagagaactc aaagaaccac cacgaggagc tcattttctt
      361 gccaaaagtt tggatgatgc cttaagactt attgaacaac cggaattggc aagtaaagta
      421 gacatggttt ggatagtcgg aggcagttct gtttaccagg aagccatgaa tcaaccaggc
      481 cacctcagac tctttgtgac aaggatcatg caggaatttg aaagtgacac gtttttccca
      541 gaaattgatt tggggaaata taaacttctc ccagaatacc caggcgtcct ctctgaggtc
      601 caggaggaaa aaggcatcaa gtataagttt gaagtctacg agaagaaaga ctaacaggaa
      661 gatgctttca agttctctgc tcccctccta aagctatgca tttttataag accatgggac
      721 ttttgctggc tttagatcat aatcagccat accacatttg tagaggtttt acttgcttta
      781 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
      841 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
      901 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
      961 tatcatgtct ggatccccgg ccaacggtct ggtgacccgg ctgcgagagc tcggtgtacc
     1021 tgagacgcga gtaagccctt gagtcaaaga cgtagtcgtt gcaagtccgc accaggtact
     1081 gatatcccac caaaaagtgc ggcggcggct ggcggtagag gggccagcgt agggtggccg
     1141 gggctccggg ggcgaggtct tccaacataa ggcgatgata tccgtagatg tacctggaca
     1201 tccaggtgat gccggcggcg gtggtggagg cgcgcggaaa gtcgcggacg cggttccaga
     1261 tgttgcgcag cggcaaaaag tgctccatgg tcgggacgct ctggccggtg aggcgtgcgc
     1321 agtcgttgac gctctagacc gtgcaaaagg agagcctgta agcgggcact cttccgtggt
     1381 ctggtggata aattcgcaag ggtatcatgg cggacgaccg gggttcgaac cccggatccg
     1441 gccgtccgcc gtgatccatg cggttaccgc ccgcgtgtcg aacccaggtg tgcgacgtca
     1501 gacaacgggg gagcgctcct tttggcttcc ttccaggcgc ggcggctgct gcgctagctt
     1561 ttttggcgag ctcgaattaa ttctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1621 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1681 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1741 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1801 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1861 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1921 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1981 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2041 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2101 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2161 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2221 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2281 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2341 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2401 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2461 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2521 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2581 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2641 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2701 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2761 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2821 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2881 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2941 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3001 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3061 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3121 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3181 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3241 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3301 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3361 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3421 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3481 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3541 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     3601 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     3661 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     3721 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     3781 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     3841 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     3901 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     3961 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     4021 aacgacggcc agtgccaagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact
     4081 acttctggaa tagctcagag gccgaggcgg cctcggcctc tgcataaata aaaaaaatta
     4141 gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg atgggcggag
     4201 ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata cttctgcctg
     4261 ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc atgctttgca
     4321 tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac acattccaca
     4381 ggatccggtc gcgcgaattt cgagcggtgt tccgcggtcc tcctcgtata gaaactcgga
     4441 ccactctgag acgaaggctc gcgtccaggc cagcacgaag gaggctaagt gggaggggta
     4501 gcggtcgttg tccactaggg ggtccactcg ctccagggtg tgaagacaca tgtcgccctc
     4561 ttcggcatca aggaaggtga ttggtttata ggtgtaggcc acgtgaccgg gtgttcctga
     4621 aggggggcta taaaaggggg tgggggcgcg ttcgtcctca ctctcttccg catcgctgtc
     4681 tgcgagggcc agctgttggg ctcgcggttg aggacaaact cttcgcggtc tttccagtac
     4741 tcttggatcg gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg acctgagcga
     4801 gtccgcatcg accggatcgg aaaacctctc gactgttggg gtgagtactc cctctcaaaa
     4861 gcgggcatga cttctgcgct aagattgtca gtttccaaaa acgaggagga tttgatattc
     4921 acctggcccg cggtgatgcc tttgagggtg gccgcgtcca tctggtcaga aaagacaatc
     4981 tttttgttgt caagcttgag gtgtggcagg cttgagatct ggccatacac ttgagtgaca
     5041 atgacatcca ctttgccttt ctctccacag gtgtccactc ccaggtccaa ctgcaggcga
     5101 gcctgaattg cggccgcact agtatggtcc gcttcgggga cgagctgggc ggccgctatg
     5161 ggggcgcggg cggcgcggag cgggcccggg gcggcggggc cggcggggct ggcggcccgg
     5221 gccccggggg gctgccgccg ggccagcggg tcctctacaa acagtcgatc gcgcaggccg
     5281 cggcgaccat ggcgctgtac aaccccatcc cggtcaagca gaactgcttc accgtcaacc
     5341 gctcgctctt cgtcttcagc gaggacaacg tcgtccgcaa atacgccaag cgcatcaccg
     5401 agtggcctta gctcgaggam wtsmksctgg ccaccatcat cgccaactgc atcgtgctgg
     5461 ccctggagca gcacctccct gacggggaca agacgcctat gtccgagcgc ctggatgaca
     5521 ccgagcccta cttcattggg atcttctgct tcgaggcggg gatcaagatc ctgncnctgg
     5581 gctttgttct gcacaagggc tcctatctgc ggaacggctg gaacgtcatg gacttcgtgg
     5641 tagtc
//
