LOCUS       sequence-202636-        9114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Calcium channel beta4 auxillary subunit in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9114)
  AUTHORS   Brodbeck J, Davies A, Courtney JM, Meir A, Balaguero N, Canti C, 
            Moss FJ, Page KM, Pratt WS, Hunt SP, Barclay J, Rees M, Dolphin AC
  TITLE     The ducky mutation in Cacna2d2 results in altered Purkinje cell 
            morphology and is associated with the expression of a truncated 
            alpha 2 delta-2 protein with abnormal function.
  JOURNAL   J Biol Chem. 2002 Mar 8;277(10):7684-93. doi: 
            10.1074/jbc.M109404200. Epub 2001 Dec 26.
  PUBMED    11756448
REFERENCE   2  (bases 1 to 9114)
  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.M109404200"; journalName: "J Biol Chem"; date: 
            "2002-03-8- 8"; volume: "277"; issue: "10"; pages: "7684-93"
COMMENT     Sequence Label: cacnb4 (rat) in pMT2 vector
FEATURES             Location/Qualifiers
     source          1..9114
                     /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"
     primer_bind     1097..1113
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1098..1114
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5162..5725
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    5913..6047
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5950..5969)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6004..6023
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6720..6737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6891..7479)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6971..6990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7650..8441,8442..8510))
                     /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     8273..8292
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8511..8615)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8683..8701
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8739..8761)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8861..8880
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9074..9096
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9088..9105
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9089..9105
                     /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 attcctcgag
     1081 ggtaccgggc cccccctcga ggtcgacggt atcgataagc ttgatatcga attggcctct
     1141 ctgggcgaat tccgggtgct ctccccaccc cgcgaggggg gaggggagcg cttcggcccc
     1201 tttaaggtgc ggcgctcggc gaagtgcggg caggcggccc gcgagccggc gcggggccag
     1261 gagccgggta gggaagtcct gcctccccct ccgcccgcac cggcccgcgc cctccgccca
     1321 ccgcgccgcg agggccatgc ggcgcctctg aaccatgtcg tcctcctacg ccaagaacgg
     1381 ggcggcggac gggccgcact cccccagctc gcaggtggcc cgaggcacca cgacccggag
     1441 gagcaggttg aaaagatccg atggcagcac cacttccacc agcttcatcc tcagacaggg
     1501 gtcagcagat tcctatacaa gcaggccatc ggactccgat gtctctttgg aagaggaccg
     1561 ggaagcgatt cggcaagaga gagaacagca agcagctatc cagcttgaga gagcgaagtc
     1621 caaacctgta gcttttgcgg tgaaaacgaa tgtgagctac tgtggtgccc tggatgagga
     1681 tgtgcccgtt cccagcacag ccatctcctt tgacgccaag gactttcttc acattaaaga
     1741 gaaatataac aatgattggt ggataggaag gctggtgaag gagggctgtg agatcggctt
     1801 catcccaagt ccactgcgtt tggagaacat acggattcaa caggaacaga aaagaggccg
     1861 ttttcatggc gggaaatcga gtggaaactc ttcctccagt ctgggagaaa tggtgtcagg
     1921 aacatttcga gcaactccca caacaacagc aaaacagaag cagaaagtga cggagcacat
     1981 tcctccgtat gatgttgtgc catcaatgcg tcctgtggtg ttagtagggc catcactgaa
     2041 aggttatgag gtaacagaca tgatgcagaa agccctcttt gatttcctga agcacaggtt
     2101 tgatgggagg atatcaataa ccagagtgac agctgacatt tctcttgcta agaggtctgt
     2161 cctcaacaat cctagcaaga gagcaataat tgaacgttca aacaccagat ccagcctagc
     2221 ggaagtacag agtgaaattg aaagaatttt cgagttggca agatctttgc aattggttgt
     2281 tctcgatgca gacaccatca atcacccagc acagctgata aagacatcgt tagcgcccat
     2341 catcgtccac gtgaaggtct cgtccccaaa ggttttacag cggctgatta agtccagagg
     2401 aaagtcccag agtaaacact tgaatgttca actggtggca gctgataaac tggcccagtg
     2461 tccccctgaa atgtttgatg ttatattaga tgagaatcaa cttgaggatg cctgtgaaca
     2521 tctgggagag tacctggagg catattggcg tgccacccac acaagcagta gcacccctat
     2581 gaccccattg ctggggagga acgtgggctc cacagctctc tcaccgtatc ccacagcaat
     2641 ctctggacta cagagtcagc ggatgaggca tagcaaccac tctacagaga attctccaat
     2701 tgaaagacga agcctaatga cctctgatga aaattaccac aatgagaggg cccgcaagag
     2761 taggaaccgc ttgtcctcca gctcccagca cagccgagat cactaccctc tggtggaaga
     2821 agattatcct gactcatacc aggacactta caaaccccat aggaaccgag gctcacctgg
     2881 gggatgcagc catgactccc gacataggct ttgaggctgc ccaaacaaac aaacaaacaa
     2941 acaaacatca tctgttgaca atttgccata gtactgctaa gataaaccag tcatcttaat
     3001 ttggcaagca tggcacagta tctgctgtgc gggtggtctg ctgtcgtttg atgtaagggg
     3061 caagaattaa atagaagtac attatgccct tgagtctagg agggtattag atacccggtc
     3121 atataggtat ttttaagtgc tacatttaca tgataacagt accttttgtt ttcttcatag
     3181 attttagaca catcaatttg taatttaggg tactgatcaa ggcacatata aaataatttc
     3241 ccgtgtagga aattccaaat gaccagttcc agttggaacc aatttataaa ccaattcctg
     3301 ttggaatttg ggtgaattga ctggaaagtc tacctgagca tgttgaaatc aattgaaaaa
     3361 atcagagaag aaaaaaatac ccaataagga aatcaaaata ctaatagaaa ccaaacccag
     3421 ctatggtatt tgtttgctgg aagctaaatt tacacttcaa gttggaatgt agaaacattt
     3481 taataatgtt gctggaatgg cttttcaaac ttaccaaata tattgatagt gccaaaaaaa
     3541 aatctcagaa atcttaatgc agaaattata taatcgcctc agattttaat atttttattt
     3601 ttcctgatat agctgttgta ttcaaagtgt tttgccttct gttcagtcag tactgttagc
     3661 cagtttgaag ttgacaaaat ccaggggata tttgagaagt ggtaaagaga agccctagca
     3721 tggacagttc cagagagtgg agaatgaggt gttgttcctt ctgcagtcag tctgcacaca
     3781 gctgcacaca tctggagacc tggtgtcagg tagcagagat gtggcggggc ctctctggct
     3841 acaacggtaa ttggcttcag atgggaagtc gtccatcgta cagcacaccg acacgggctt
     3901 aggagctgtg ccagcatggc tgagtgtctt tccctcactt cagccagtct ccatctttga
     3961 atataatgaa tgttttggcc ctgcatcaca cttcttttat gtttcatggg cataatcaaa
     4021 ttaggtactg aaagttatgt tttcacaaaa taattaggaa aacttaaaac actgaattaa
     4081 aatcttgcag agaaagaaaa aaaaaccctc atagaatcag aatatattta ttcactaaaa
     4141 gcaagacagt ttcaatacac ggcattagag agagttgaga gactgctcgg ttccattgat
     4201 agtgaacagc ctttagactt ctatgagtgt atttatggaa ttgatgacct atgtcctatg
     4261 ttattaaaga cagctttaat ccaagaaatt agtgaaattg accctttgaa acaaatgtct
     4321 ttctgaaagg gcttgtttct gaaatgacgc aatacatgtg ttttctaaag acccaattgg
     4381 agctagacgt aaagtctctt tctactaatg ctttgaagtc aatttccgaa acaagtataa
     4441 ttaatagtat gttaatttct ccaaaaaaaa gacatgggtc ctcaaaactg tgtaccagtt
     4501 cattaattct cctgatcaag agtctaagtt tcatattctt aatcagttat cctatctcgg
     4561 atcccatcaa cccattgtct tgactgtatg tgcaacccat tatgaacctg cttatctgat
     4621 aatgaggctg cctgaaattt tgtggtaatc tcaaacattt gatacattta aacacgggct
     4681 cagtcaaatt gacaagctaa catctgacac tggcttgtga agcttttttg tcatccagcg
     4741 aggcttacag aattttctgt taagacattt aacagagctg tattcattgg gctctttaat
     4801 aagcatataa tcttccatta ccaaatactt tgttagaaga cagaagagca tttttccagc
     4861 gaacaaacta tatttattac atgtatagca catatttgca tgagaaagaa aaacattagc
     4921 caaacatttt cctgtatgct ttactggttt cttgtttgtg cgcattaaag catttaattg
     4981 caaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaggccggag
     5041 cggccgcgtc gacgaattcc gggggggggg ggggggggga cagctcaggg ctgcgatttc
     5101 gcgccaaact tgacggcaat cctagcgtga aggctggtag gattttatcc ccgctgccat
     5161 catggttcga ccattgaact gcatcgtcgc cgtgtcccaa aatatgggga ttggcaagaa
     5221 cggagaccta ccctggcctc cgctcaggaa cgagttcaag tacttccaaa gaatgaccac
     5281 aacctcttca gtggaaggta aacagaatct ggtgattatg ggtaggaaaa cctggttctc
     5341 cattcctgag aagaatcgac ctttaaagga cagaattaat atagttctca gtagagaact
     5401 caaagaacca ccacgaggag ctcattttct tgccaaaagt ttggatgatg ccttaagact
     5461 tattgaacaa ccggaattgg caagtaaagt agacatggtt tggatagtcg gaggcagttc
     5521 tgtttaccag gaagccatga atcaaccagg ccacctcaga ctctttgtga caaggatcat
     5581 gcaggaattt gaaagtgaca cgtttttccc agaaattgat ttggggaaat ataaacttct
     5641 cccagaatac ccaggcgtcc tctctgaggt ccaggaggaa aaaggcatca agtataagtt
     5701 tgaagtctac gagaagaaag actaacagga agatgctttc aagttctctg ctcccctcct
     5761 aaagctatgc attttttata agaccatggg acttttgctg gctttagatc ataatcagcc
     5821 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     5881 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     5941 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     6001 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcccc ggccaacggt
     6061 ctggtgaccc ggctgcgaga gctcggtgta cctgagacgc gagtaagccc ttgagtcaaa
     6121 gacgtagtcg ttgcaagtcc gcaccaggta ctgatatccc accaaaaagt gcggcggcgg
     6181 ctggcggtag aggggccagc gtagggtggc cggggctccg ggggcgaggt cttccaacat
     6241 aaggcgatga tatccgtaga tgtacctgga catccaggtg atgccggcgg cggtggtgga
     6301 ggcgcgcgga aagtcgcgga cgcggttcca gatgttgcgc agcggcaaaa agtgctccat
     6361 ggtcgggacg ctctggccgg tgaggcgtgc gcagtcgttg acgctctaga ccgtgcaaaa
     6421 ggagagcctg taagcgggca ctcttccgtg gtctggtgga taaattcgca agggtatcat
     6481 ggcggacgac cggggttcga accccggatc cggccgtccg ccgtgatcca tccggttacc
     6541 gcccgcgtgt cgaacccagg tgtgcgacgt cagacaacgg gggagcgctc cttttggctt
     6601 ccttccaggc gcggcggctg ctgcgctagc ttttttggcg agctcgaatt aattctgcat
     6661 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6721 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6781 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6841 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6901 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6961 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     7021 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     7081 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     7141 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     7201 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     7261 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     7321 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     7381 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7441 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7501 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7561 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7621 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7681 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7741 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7801 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7861 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7921 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7981 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     8041 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     8101 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     8161 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     8221 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     8281 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     8341 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     8401 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8461 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8521 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8581 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8641 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     8701 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     8761 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     8821 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     8881 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     8941 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     9001 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     9061 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gctt
//
