LOCUS       sequence-415909-        8902 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of rat alpha2delta-1 calcium channel auxillary subunit 
            and with R421A mutation that disrupts Gabapentin binding.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8902)
  AUTHORS   Cassidy JS, Ferron L, Kadurin I, Pratt WS, Dolphin AC
  TITLE     Functional exofacially tagged N-type calcium channels elucidate the 
            interaction with auxiliary alpha2delta-1 subunits.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Jun 17;111(24):8979-84. doi: 
            10.1073/pnas.1403731111. Epub 2014 Jun 2.
  PUBMED    24889613
REFERENCE   2  (bases 1 to 8902)
  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: 
            "10.1073/pnas.1403731111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-06-17- 17"; volume: "111"; issue: "24"; pages: "8979-84"
COMMENT     Sequence Label: cacna2d1 R241A pMT2
FEATURES             Location/Qualifiers
     source          1..8902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             93..656
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    843..977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(880..899)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     934..953
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1650..1667)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1821..2409)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1901..1920)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2580..3371,3372..3440))
                     /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     3203..3222
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3441..3545)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3613..3631
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3669..3691)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3791..3810
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4004..4026
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4018..4035
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4019..4035
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4045..4374)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4059..4194
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4359..4379
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          4843..5072
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     5055..5074
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 gggggggggg acagctcagg gctgcgattt cgcgccaaac ttgacggcaa tcctagcgtg
       61 aaggctggta ggattttatc cccgctgcca tcatggttcg accattgaac tgcatcgtcg
      121 ccgtgtccca aaatatgggg attggcaaga acggagacct accctggcct ccgctcagga
      181 acgagttcaa gtacttccaa agaatgacca caacctcttc agtggaaggt aaacagaatc
      241 tggtgattat gggtaggaaa acctggttct ccattcctga gaagaatcga cctttaaagg
      301 acagaattaa tatagttctc agtagagaac tcaaagaacc accacgagga gctcattttc
      361 ttgccaaaag tttggatgat gccttaagac ttattgaaca accggaattg gcaagtaaag
      421 tagacatggt ttggatagtc ggaggcagtt ctgtttacca ggaagccatg aatcaaccag
      481 gccacctcag actctttgtg acaaggatca tgcaggaatt tgaaagtgac acgtttttcc
      541 cagaaattga tttggggaaa tataaacttc tcccagaata cccaggcgtc ctctctgagg
      601 tccaggagga aaaaggcatc aagtataagt ttgaagtcta cgagaagaaa gactaacagg
      661 aagatgcttt caagttctct gctcccctcc taaagctatg catttttata agaccatggg
      721 acttttgctg gctttagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
      781 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
      841 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
      901 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
      961 cttatcatgt ctggatcccc ggccaacggt ctggtgaccc ggctgcgaga gctcggtgta
     1021 cctgagacgc gagtaagccc ttgagtcaaa gacgtagtcg ttgcaagtcc gcaccaggta
     1081 ctgatatccc accaaaaagt gcggcggcgg ctggcggtag aggggccagc gtagggtggc
     1141 cggggctccg ggggcgaggt cttccaacat aaggcgatga tatccgtaga tgtacctgga
     1201 catccaggtg atgccggcgg cggtggtgga ggcgcgcgga aagtcgcgga cgcggttcca
     1261 gatgttgcgc agcggcaaaa agtgctccat ggtcgggacg ctctggccgg tgaggcgtgc
     1321 gcagtcgttg acgctctaga ccgtgcaaaa ggagagcctg taagcgggca ctcttccgtg
     1381 gtctggtgga taaattcgca agggtatcat ggcggacgac cggggttcga accccggatc
     1441 cggccgtccg ccgtgatcca tgcggttacc gcccgcgtgt cgaacccagg tgtgcgacgt
     1501 cagacaacgg gggagcgctc cttttggctt ccttccaggc gcggcggctg ctgcgctagc
     1561 ttttttggcg agctcgaatt aattctgcat taatgaatcg gccaacgcgc ggggagaggc
     1621 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1681 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1741 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1801 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1861 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1921 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     1981 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2041 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2101 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2161 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2221 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2281 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2341 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2401 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2461 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     2521 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     2581 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     2641 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     2701 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     2761 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     2821 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     2881 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     2941 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3001 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3061 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3121 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3181 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3241 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3301 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3361 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3421 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     3481 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     3541 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     3601 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     3661 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     3721 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     3781 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     3841 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     3901 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     3961 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     4021 aaaacgacgg ccagtgccaa gctttttgca aaagcctagg cctccaaaaa agcctcctca
     4081 ctacttctgg aatagctcag aggccgaggc ggcctcggcc tctgcataaa taaaaaaaat
     4141 tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg ggatgggcgg
     4201 agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca tacttctgcc
     4261 tgctggggag cctggggact ttccacacct ggttgctgac taattgagat gcatgctttg
     4321 catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac acacattcca
     4381 caggatccgg tcgcgcgaat ttcgagcggt gttccgcggt cctcctcgta tagaaactcg
     4441 gaccactctg agacgaaggc tcgcgtccag gccagcacga aggaggctaa gtgggagggg
     4501 tagcggtcgt tgtccactag ggggtccact cgctccaggg tgtgaagaca catgtcgccc
     4561 tcttcggcat caaggaaggt gattggttta taggtgtagg ccacgtgacc gggtgttcct
     4621 gaaggggggc tataaaaggg ggtgggggcg cgttcgtcct cactctcttc cgcatcgctg
     4681 tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt
     4741 actcttggat cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc
     4801 gagtccgcat cgaccggatc ggaaaacctc tcgactgttg gggtgagtac tccctctcaa
     4861 aagcgggcat gacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat
     4921 tcacctggcc cgcggtgatg cctttgaggg tggccgcgtc catctggtca gaaaagacaa
     4981 tctttttgtt gtcaagcttg aggtgtggca ggcttgagat ctggccatac acttgagtga
     5041 caatgacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcaggc
     5101 gagcctgaat tcctcgaggg taccgagctc ggatccacta gtaacggccg ccagtgtgct
     5161 ggaattctgc agatcgcgaa gatggctgct ggctgcctgc tggccttgac tctgacactt
     5221 ttccaatctt ggctgatcgg cccctcgagc gaggagccct tcccttcgcc cgtcactatc
     5281 aagtcatggg tggataaaat gcaggaagac cttgtcacac tggcaaagac agcaagtgga
     5341 gtgactcagc ttgctgacat ttatgaaaaa taccaagatt tgtatactgt ggagccaaat
     5401 aatgcacgcc aactggttga aattgcagct agagacattg agaaacttct gagcaacaga
     5461 tctaaagccc tggtgcgcct ggctatggaa gcagagaaag tccaagccgc ccaccaatgg
     5521 agggaagatt ttgcaagcaa cgaagttgtc tactacaacg ctaaagacga tcttgatcct
     5581 gaaagaaatg agagtgagtc aggcagccaa cggattaaac ctgttttcat cgaagacgct
     5641 aattttggac gccagatatc ctatcaacat gcagcggtcc atattcccac ggacatctat
     5701 gagggctcaa ccatagtgtt aaacgaactc aactggacaa gtgccttaga tgaagttttc
     5761 aaaagaaatc gagatgaaga ccctacattg ctgtggcaag tgtttggcag cgccaccggc
     5821 ctggcccgat attatccagc ttctccgtgg gtggataaca gcagaactcc aaacaagatc
     5881 gatctctacg atgtacgcag agcaccatgg tacatccagg gagctgcatc ccccaaagac
     5941 atgctcattc tggtggacgt gagtggaagt gtgagcggat tgactctgaa actcatccgg
     6001 acatctgtct ctgagatgtt agaaaccctt tccgatgatg atttcgtgaa tgtagcttca
     6061 tttaacagca atgctcagga tgtgagttgt ttccagcacc tggttcaagc gaatgtaagg
     6121 aacaagaagg tgctgaagga tgcggtgaat aacatcacag caaaggggat aacagattac
     6181 aagaaaggct ttagcttcgc gtttgaacag ctacttaatt ataatgtttc cagagccaat
     6241 tgtaataaga tcatcatgtt attcacggat ggaggagaag agagagccca ggagatattt
     6301 gccaagtaca ataaagacaa aaaagtccgc gtgtttacat tttctgttgg tcaacataat
     6361 tatgacagag gacctattca gtggatggct tgtgaaaaca aaggttatta ttatgagatt
     6421 ccctctattg gtgcaataag aatcaatact caggaatatc tagatgttct gggaagacca
     6481 atggttttag ccggggacaa agcaaagcaa gtccagtgga caaatgtgta tttggatgct
     6541 ctggagctgg gacttgtcat tactggaact ctaccggtct tcaacgtcac tggccagtct
     6601 gaaaataaga ctaacttgaa gaaccagttg attcttggtg tgatgggcgt tgatgtgtct
     6661 ctggaagata ttaagagatt gacaccacgt tttacactgt gccccaacgg ttactatttt
     6721 gcaatcgatc ctaatggtta tgtcttactg catccaaatc ttcagccaaa gaaccccaaa
     6781 tctcaggagc cggtcacact ggattttctc gatgctgagt tagagaatga tattaaagtg
     6841 gagattcgaa ataaaatgat agacggagaa agtggagaaa aaacattcag aactctggtc
     6901 aaatctcaag atgagagata tattgacaaa ggaaatcgaa catacacatg gacacctgtc
     6961 aatggcacag attacagttt ggccttggta ttgccaacct acagttttta ctatataaaa
     7021 gccaaaatag aagagacaat aactcaggcc agatcaaaaa aggggaaaat gaaggattca
     7081 gaaaccctga agccagacaa ttttgaagaa tctggctaca ctttcatagc accaagggaa
     7141 tactgcaatg acctgaaacc ttcagataat aacactgaat ttcttttaaa tttcaatgaa
     7201 tttattgata ggaaaactcc aaacaaccct tcctgtaata cagatttgat taatagaatc
     7261 ttgctggatg cgggttttac aaatgaactt gtccaaaatt attggagtaa acagaaaaat
     7321 atcaaaggag tgaaggcacg ctttgtggtg actgatggcg gaattacaag agtttacccc
     7381 aaagaggctg gagaaaactg gcaagagaac ccagagacgt atgaggacag cttctacaaa
     7441 cggagcctag ataacgataa ctacgttttc actgcgccct acttcaacaa aagtggacct
     7501 ggtgcctatg aatctggaat tatggtaagc aaagctgtag aactgtacat ccaaggaaaa
     7561 cttcttaagc ctgcagttgt gggaattaaa attgacgtaa actcctggat agaaaatttt
     7621 accaaaactt caatcaggga tccgtgtgct ggtccagttt gtgactgcaa aagaaacagt
     7681 gatgtaatgg actgtgtcat tctagatgat ggtggatttc ttctgatggc caatcatgat
     7741 gattacacta atcagattgg acgttttttt ggagagattg acccgagcat gatgagacac
     7801 ctggttaata tatcacttta tgcattcaat aaatcatatg actatcagtc tgtgtgtgat
     7861 ccaggggcag caccaaagca aggagcagga catcgctcag cctatgtgcc atcaattaca
     7921 gacatactcc agattggctg gtgggccact gctgccgcct ggtctattct ccagcagcta
     7981 ctcttgagtt tgacatttcc acggctcctt gaggcagttg aaatggaaga ggatgacttc
     8041 acagcttccc tgtctaagca gagctgcatc acagaacaaa ctcagtactt cttcaagaac
     8101 gatactaaat cattcagtgg tttactggac tgtggaaact gttccaggat ctttcatgtt
     8161 gagaagctta tgaacaccaa cctagtgttc ataatggtgg agagcaaggg aacatgtccg
     8221 tgtgacacgc ggctgctcat gcaagcagaa cagacttctg atggtccaga tccttgcgac
     8281 atggttaagc agcccagata ccgaaaagga cctgatgtct gctttgataa caatgtgctg
     8341 gaggattata ccgactgtgg tggtgtttct ggattaaacc cttccttatg gtctatcttt
     8401 ggactccaat ttatactcct ttggctggta tctggcagca gacactatct atggtgaccc
     8461 tctaaaccca tgatctgcaa cttaattcca tgccctgcca aaacatgagc cctcagttac
     8521 attaaaatag agcccgttga agaagctcca cagaacctta gctgggcctc taccctgatg
     8581 taactgtgag gcttagattc ctgaccatcg ctggctgcat gtcagggtgt tggaaactga
     8641 aactcgtgtg aacatgctgc atcatctgtg tccaacatca aagcaaagct ctactcattc
     8701 tctgtgggag gtttgggagt tcgctgttgc actgttggag attacgtgga aaaggctccc
     8761 ccacgcaggc cttatgaaaa gaagcagtct tgactttgac ctagcatttc aacctgctgc
     8821 agttttacca agccgaattc ctgcagcccg ggggatccac tagatccatc acactggcgg
     8881 ccgcgtcgac gaattccggg gg
//
