LOCUS       sequence-415911-        8996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of rabbit Cav2.1 calcium channel with a CaV2.1-P1217fs 
            mutation causing a truncated protein, (EA2). R57A R59A mutations 
            prevent dominant-negative suppression of Cav2.1 currents .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8996)
  AUTHORS   Dahimene S, Page KM, Nieto-Rostro M, Pratt WS, D'Arco M, Dolphin AC
  TITLE     A CaV2.1 N-terminal fragment relieves the dominant-negative 
            inhibition by an Episodic ataxia 2 mutant.
  JOURNAL   Neurobiol Dis. 2016 Sep;93:243-56. doi: 10.1016/j.nbd.2016.05.020. 
            Epub 2016 May 31.
  PUBMED    27260834
REFERENCE   2  (bases 1 to 8996)
  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: "Neurobiol 
            Dis."; date: "2016-09"; volume: "93:243-56. doi"; pages: " 
            10.1016/j.nbd.2016.05.020. Epub 2016 May 31"
COMMENT     Sequence Label: Cav2.1 EA2, R57A, R59A  pMT2
FEATURES             Location/Qualifiers
     source          1..8996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             190..753
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    940..1074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(977..996)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1031..1050
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1747..1764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1918..2506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1998..2017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2677..3468,3469..3537))
                     /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     3300..3319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3538..3642)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3710..3728
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3766..3788)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3888..3907
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4101..4123
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4115..4132
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4116..4132
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4142..4471)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4156..4291
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4456..4476
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          4940..5169
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     5152..5171
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 gggggggggg gggggggrvv bbbbvrkssk kbkdkkkbbs svvvvvdbkb rvssvvvvbb
       61 vbssvssrrs sssssssdkb kddddbvbss vrvdvddkrr rrvrrksaca gctcagggct
      121 gcgatttcgc gccaaacttg acggcaatcc tagcgtgaag gctggtagga ttttatcccc
      181 gctgccatca tggttcgacc attgaactgc atcgtcgccg tgtcccaaaa tatggggatt
      241 ggcaagaacg gagacctacc ctggcctccg ctcaggaacg agttcaagta cttccaaaga
      301 atgaccacaa cctcttcagt ggaaggtaaa cagaatctgg tgattatggg taggaaaacc
      361 tggttctcca ttcctgagaa gaatcgacct ttaaaggaca gaattaatat agttctcagt
      421 agagaactca aagaaccacc acgaggagct cattttcttg ccaaaagttt ggatgatgcc
      481 ttaagactta ttgaacaacc ggaattggca agtaaagtag acatggtttg gatagtcgga
      541 ggcagttctg tttaccagga agccatgaat caaccaggcc acctcagact ctttgtgaca
      601 aggatcatgc aggaatttga aagtgacacg tttttcccag aaattgattt ggggaaatat
      661 aaacttctcc cagaataccc aggcgtcctc tctgaggtcc aggaggaaaa aggcatcaag
      721 tataagtttg aagtctacga gaagaaagac taacaggaag atgctttcaa gttctctgct
      781 cccctcctaa agctatgcat ttttataaga ccatgggact tttgctggct ttagatcata
      841 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
      901 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
      961 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     1021 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatccccggc
     1081 caacggtctg gtgacccggc tgcgagagct cggtgtacct gagacgcgag taagcccttg
     1141 agtcaaagac gtagtcgttg caagtccgca ccaggtactg atatcccacc aaaaagtgcg
     1201 gcggcggctg gcggtagagg ggccagcgta gggtggccgg ggctccgggg gcgaggtctt
     1261 ccaacataag gcgatgatat ccgtagatgt acctggacat ccaggtgatg ccggcggcgg
     1321 tggtggaggc gcgcggaaag tcgcggacgc ggttccagat gttgcgcagc ggcaaaaagt
     1381 gctccatggt cgggacgctc tggccggtga ggcgtgcgca gtcgttgacg ctctagaccg
     1441 tgcaaaagga gagcctgtaa gcgggcactc ttccgtggtc tggtggataa attcgcaagg
     1501 gtatcatggc ggacgaccgg ggttcgaacc ccggatccgg ccgtccgccg tgatccatgc
     1561 ggttaccgcc cgcgtgtcga acccaggtgt gcgacgtcag acaacggggg agcgctcctt
     1621 ttggcttcct tccaggcgcg gcggctgctg cgctagcttt tttggcgagc tcgaattaat
     1681 tctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     1741 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1801 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1861 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1921 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1981 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2041 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2101 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     2161 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     2221 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     2281 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     2341 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     2401 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     2461 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     2521 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2581 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2641 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2701 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2761 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2821 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2881 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2941 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3001 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3061 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3121 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     3181 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     3241 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     3301 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     3361 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     3421 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     3481 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3541 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3601 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3661 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3721 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3781 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3841 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     3901 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     3961 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     4021 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     4081 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct
     4141 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     4201 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     4261 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
     4321 ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc
     4381 cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc
     4441 ctggggactt tccacaccct aactgacaca cattccacag gatccggtcg cgcgaatttc
     4501 gagcggtgtt ccgcggtcct cctcgtatag aaactcggac cactctgaga cgaaggctcg
     4561 cgtccaggcc agcacgaagg aggctaagtg ggaggggtag cggtcgttgt ccactagggg
     4621 gtccactcgc tccagggtgt gaagacacat gtcgccctct tcggcatcaa ggaaggtgat
     4681 tggtttatag gtgtaggcca cgtgaccggg tgttcctgaa ggggggctat aaaagggggt
     4741 gggggcgcgt tcgtcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc
     4801 tcgcggttga ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg
     4861 gcctccgaac ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga
     4921 aaacctctcg actgttgggg tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta
     4981 agattgtcag tttccaaaaa cgaggaggat ttgatattca cctggcccgc ggtgatgcct
     5041 ttgagggtgg ccgcgtccat ctggtcagaa aagacaatct ttttgttgtc aagcttgagg
     5101 tgtggcaggc ttgagatctg gccatacact tgagtgacaa tgacatccac tttgcctttc
     5161 tctccacagg tgtccactcc caggtccaac tgcaggcgag cctgaattcc tcgaggtctt
     5221 gcagaatggc ccgctttgga gacgagatgc cgggccgcta cggcgcaggc ggaggaggct
     5281 cagggccggc cgccggggtg gtcgtgggcg ccgcgggcgg ccgaggagcc gggggcagcc
     5341 ggcagggcgg gcagcccgga gcgcagagga tgtacaagca gtcgatggcg caggcagcgg
     5401 cgaccatggc cctctacaac cccatccctg tccgccagaa ctgcctcacg gtcaaccgct
     5461 ccctgttcct cttcagtgaa gacaacgtgg tgagaaaata cgccaaaaag atcacggaat
     5521 ggcctccctt cgagtacatg atcctggcca ccatcattgc taactgcatc gtcctggccc
     5581 tggagcagca cctccctgat gatgacaaga cacccatgtc cgagcggctg gatgacacag
     5641 aaccctattt cattggcatc ttctgttttg aggctggaat taagatcgtg gctcttggct
     5701 ttgccttcca caaaggctcc tacctgagga atggctggaa cgtcatggac tttgtcgtgg
     5761 tgctaacagg catcttggcc actgtcggga cggagtttga tctacggaca ctgagggcgg
     5821 ttcgtgtgct gcggccactc aagctggtgt ctggaatccc aagtttacaa gtcgtcctga
     5881 agtcaatcat gaaggcgatg atccctctgc tgcagatcgg cctcctcctg ttttttgcaa
     5941 tccttatttt tgcaatcata gggttagaat tttatatggg aaaatttcat accacctgct
     6001 ttgaagaggg gacagacgac atccagggtg agtcgccagc tccgtgtggg acagaggagc
     6061 ctgcccgcac ctgccccaac gggaccaaat gtcagccgta ctgggaaggg cccaacaacg
     6121 gcatcactca gttcgacaac atcctgtttg ctgtgctcac tgttttccag tgcatcacca
     6181 tggaaggctg gactgatctc ctctacaata gcaacgatgc ctcagggaac acttggaact
     6241 ggttgtactt catccccctc atcatcatcg gctccttttt tatgctgaac cttgtgctgg
     6301 gtgtgctgtc tggggagttt gccaaagaaa gggaacgtgt agagaaccga agggcttttc
     6361 tgaagctcag aagacaacag cagattgaac gtgagctcaa tggatacatg gagtggatct
     6421 cgaaagcaga agaggtgatc ctcgcggagg acgaaacaga cgtggagcag aggcaccctt
     6481 ttgatggagc tcttcggaga gctactctga agaaaagcaa gacggacctg ctcaaccctg
     6541 aggaggcgga ggaccagctt gctgacatcg cgtctgtggg gtctcccttc gccagagcca
     6601 gcatcaaaag tgccaagctg gagaattcga cttttttcca caaaaaggag agaagaatgc
     6661 gtttctacat ccgccgcatg gtcaaaactc aggccttcta ctggaccgtg ctcagtctgg
     6721 tagccctcaa cacgctgtgc gtcgccattg tccactacaa ccagcccgag tggctctccg
     6781 acttcctcta ctatgcagaa ttcattttct taggactctt tatgtccgaa atgtttataa
     6841 aaatgtatgg gctcgggaca cggccttact tccactcttc cttcaactgc tttgactgtg
     6901 gggtcatcat cgggagcatc tttgaagtca tctgggccgt catcaaaccg ggtacatcct
     6961 ttggaatcag cgtgttacga gctctcaggt tactgcgtat tttcaaagtc acaaagtact
     7021 gggcatctct cagaaacctg gttgtctccc tcctcaactc catgaaatcc atcataagtc
     7081 tgctgttcct cctcttcctc ttcattgtcg tctttgccct cttggggatg cagctgtttg
     7141 gtggccagtt taattttgac gaggggactc ctcccaccaa cttcgacact tttccagcag
     7201 caataatgac tgtgtttcag atcctgactg gcgaggattg gaatgaggtc atgtatgatg
     7261 ggatcaagtc tcaggggggc gtgcagggcg gcatggtgtt ctccatctac ttcatcgtcc
     7321 tcaccctctt cgggaactac accctgctga acgtgttctt ggctatcgcg gtggacaacc
     7381 tggccaacgc ccaggaactc accaaggatg aacaagaaga ggaagaggca gccaatcaga
     7441 aactggctct acagaaagcc aaggaggtgg cagaagtgag tcccctgtct gcagccaaca
     7501 tgtccatagc tgtgaaggaa cagcagaaga accagaagcc tgccaagtcg gtgtgggagc
     7561 agcgcaccag cgagatgcgc aagcagaacc tgctggctag ccgcgaggcg ctgtacgggg
     7621 acgcggctga gcgctggccc accacttacg cgcgcccgct gcggccggac gtgaagacgc
     7681 acttggaccg gccgctcgtg gtggacccgc aggagaaccg taacaacaac accaacaaga
     7741 gccgtgcgcc agaagcgctg cgccaaaccg cgcggccccg cgagagcgcg cgcgaccccg
     7801 acgcgcggcg cgcctggccc agcagccctg agcgcgcccc tggacgagag ggcccgtatg
     7861 gccgcgagag cgagccgcaa cagcgcgagc acgcgccacc ccgcgagcac gtaccctggg
     7921 acgcggatcc tgagcgcgcc aaggccgggg acgcgccccg ccgccacacg caccggcctg
     7981 tggccgaggg cgagcctcgt cgccaccgcg cgcgccgccg gcccggggac gaaccggacg
     8041 acagaccgga gcgcaggccg cgtccccgcg acgccactag gccggcccgc gctgcagacg
     8101 gcgaaggcga tgatggggag cgcaagcggc gacaccgaca cgggccgccg gcccacgatg
     8161 acagggagcg cagacaccgg cgcagaaaag agagccaggg ctctggggtc cccatgtctg
     8221 gtcccaacct gtccaccacc aggccaatcc agcaggatct gggccgccag gacctgccac
     8281 tggctgagga cctggacaac atgaagaaca acaagttggc caccggggag cctgccagtc
     8341 cccacgacag cctgggccac agtggccttc cccctagccc tgccaagatc gggaacagca
     8401 ccaaccctgg tcccgccttg gccaccaatc cccagaatgc tgccagccgc aggacgccca
     8461 acaacccggg caacccgtcc aaccccggcc cccccaagac tcccgagaac agccttatcg
     8521 tcaccaaccc cagcagcacc cagcccaact cagcaaagac tgccaggaaa cccgagcaca
     8581 tggcggtgga gatccccccg gcctgcccgc ccctcaacca cactgtggtc caagtaaaca
     8641 aaaacgccaa cccagaccca ctgccaaaga aagaggaaga gaagaaggag gaagaggagg
     8701 cagaccccgg ggaggatggc ccaaagccca tgccgcccta cagctccatg ttcatcctct
     8761 ccaccaccaa cccccttcgc cggctgtgcc attacatcct gaacctgcgc tacttcgaga
     8821 tgtgcatcct catggtcatt gccatgagta gcatcgcgct ggccgccgag gaccggtgca
     8881 gcccaacgca ccccgcaaca acgtgctgcg atattttgac tatgttttca caggagtgtt
     8941 tacctttgag atggtgatca agatgatcga ctagtgcggc cgcgtcgacg aattcc
//
