LOCUS       sequence-176913-        8646 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Cav2.2 DomI and DomII, this contains the 
            first 2 domains (amino acids 1 -1154) of Cav2.2 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8646)
  AUTHORS   Raghib A, Bertaso F, Davies A, Page KM, Meir A, Bogdanov Y, Dolphin 
            AC
  TITLE     Dominant-negative synthesis suppression of voltage-gated calcium 
            channel Cav2.2 induced by truncated constructs.
  JOURNAL   J Neurosci. 2001 Nov 1;21(21):8495-504.
  PUBMED    11606638
REFERENCE   2  (bases 1 to 8646)
  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: "J 
            Neurosci."; date: "2001-11-1"; volume: "21(21)"; pages: "8495-504"
COMMENT     Sequence Label: pMT2 Cav2.2 DomI and DomII
FEATURES             Location/Qualifiers
     source          1..8646
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3586..4149
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    4336..4470
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4373..4392)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4427..4446
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5143..5160)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5314..5902)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5394..5413)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6073..6864,6865..6933))
                     /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     6696..6715
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6934..7038)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7106..7124
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7162..7184)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7284..7303
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7497..7519
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7511..7528
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7512..7528
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7538..7867)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7552..7687
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7852..7872
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          8336..8565
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     8548..8567
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 atggtccgct tcggggacga gctgggcggc cgctatgggg gcgcgggcgg cgcggagcgg
       61 gcccggggcg gcggggccgg cggggctggc ggcccgggcc ccggggggct gccgccgggc
      121 cagcgggtcc tctacaaaca gtcgatcgcg cagcgcgcgc ggaccatggc gctgtacaac
      181 cccatcccgg tcaagcagaa ctgcttcacc gtcaaccgct cgctcttcgt cttcagcgag
      241 gacaacgtcg tccgcaaata cgccaagcgc atcaccgagt ggcctccgtt cgagtacatg
      301 atcctggcca ccatcatcgc caactgcatc gtgctggccc tggagcagca cctccctgac
      361 ggggacaaga cgcctatgtc cgagcgcctg gatgacaccg agccctactt cattgggatc
      421 ttctgcttcg aggcggggat caagatcctg gccctgggct ttgttctgca caagggctcc
      481 tatctgcgga acggctggaa cgtcatggac ttcgtggtag tcctcacggg gattcttgcc
      541 acagccggaa ccgacttcga cctgcgaacg ctgagggctg tgcgtgtgct aaggcccctg
      601 aagctggtgt ctgggattcc gagcttgcag gtggtgctca agtccatcat gaaggccatg
      661 gtcccgctgc tgcagatcgg gctgctgctc ttcttcgcca tcctcatgtt tgccatcatc
      721 ggcctcgagt tctacatggg caagttccac aaggcctgct tccccaacag cacagaccca
      781 gatcccgtgg gtgacttccc ctgtggcaag gaggccccgg cccggctctg tgagggagac
      841 acggagtgcc gggagtactg ggccggaccc aactttggta tcaccaactt tgacaacatc
      901 ctgtttgcca tcttgaccgt gttccagtgc atcaccatgg agggctggac tgacatcctc
      961 tacaatacaa acgatgcagc cggcaacacg tggaactggc tctacttcat ccctctcatc
     1021 atcatcggct ccttcttcat gctcaacctg gtgctgggtg tgctctcagg agagtttgct
     1081 aaggagcgcg agagagtgga gaaccgccgc gccttcctga agctccggcg gcagcagcag
     1141 attgagcggg agctcaacgg gtacctggag tggatcttca aggctgagga agtcatgctg
     1201 gctgaggagg acaggaacgc agaggagaag tcccctctgg atgcagtgtt gaagagagca
     1261 gctgccaaga agagcagaag cgacctgatc caggccgagg agggggaggg ccggctcaca
     1321 ggcctctgtg ctcccggttc cccgttcgcc cgtgccagcc tgaaaagcgg gaagacggag
     1381 agctcgtcgt acttccggag gaaggagaag atgttccgct tcttcatccg gcgcatggtg
     1441 aaggcacaga gcttctactg gacggtgctg tgtgtggtgg cactgaacac actgtgcgtg
     1501 gccatggtgc actacaacca gccgcagcgg ctcaccacgg cattgtactt tgcagagttt
     1561 gttttcctgg gtctcttcct cacagagatg tccctgaaga tgtatggcct ggggcccagg
     1621 agctacttcc ggtcctcctt caactgtttt gactttgggg tcatcgtggg cagtatcttt
     1681 gaagtggtct gggctgctgt caagccaggg acctccttcg ggatcagtgt tctgcgggct
     1741 ctgcgcttac tgagaatctt caaggtcacc aaatactgga actccctacg gaacctggtg
     1801 gtgtctctgc tcaactccat gaagtccatc atcagcctcc tcttcctgct cttcctgttc
     1861 attgttgtct ttgctctgct ggggatgcag ctgtttggag gccagttcaa ctttaaagac
     1921 gagaccccaa cgaccaactt tgacaccttc cctgctgcta tcctcaccgt cttccagatc
     1981 ctgaccggag aggactggaa tgcggtcatg taccacggga tcgagtcgca aggtggcgtc
     2041 agcagaggaa tgttctcctc cttctacttc attgtcctca ctctgtttgg gaactacacc
     2101 ctgctgaatg tcttcctggc cattgctgtg gacaaccttg ccaatgctca ggagctgacc
     2161 aaggatgaag aggagatgga agaagcagcc aatcagaaac tcgccttgca gaaggccaag
     2221 gaagtggctg aagtcagccc catgtctgct gcgaacatct ccattgccgc caggcagcag
     2281 aactcggcca aagcgcgctc ggtgtgggag cagcgagcca gccagctgcg gctgcagaac
     2341 ctgagggcca gctgcgaggc gctgtacagc gagatggacc ccgaggagag gctgcgctac
     2401 gccaccgcgc gccacctgcg gcccgacgtg aagacgcacc tggaccggcc gctggtggtt
     2461 gagcccggcc gcgacgctcc gcggggcccc ccggggggga agtcccggcc ggatggctcg
     2521 gaggcccccg agggtgcgga tcccccgcgc aggcatcacc ggcaccgaga caaggacaag
     2581 gctcccgcca cggtcccctc ggcgggggaa caggacagag ccgaagctct gagggcagag
     2641 ggcggggagc tggggccccg cgaggagcgg gggcggccac gccgcagccg cagcaaggag
     2701 gccccgggcg ccccggaggt gcgcagcgat cgcggccgtg gcccgtgtcc tgaaggtggt
     2761 cggcgccacc accggcgcgg gtcaccggag gaggcagctg agagggagcc ccgccgccac
     2821 cgcgcgcacc ggcacgggcc cgacccgggc aaggagggcc cagcgtccgg gaccaggggc
     2881 gagcggcgcg cacgacaccg cacgggcccc cgggcgtgcc cccgggaggc tgagagcagc
     2941 gaggagcctg cgcggcggca ccgggctcgg cacaaggcgc cgcccaccca ggagaccgcg
     3001 gagaaggaca aggaggctgc cgagaagggg ggcgaggcca cggaggccga gaaggacaag
     3061 gaggctcgga accaccagcc caaggagctg ccctgtgacc tggaggccat cggcatgctg
     3121 ggagtgggcg ctgtgcatac gctgcccagc acctgtctgc agaaggtgga ggagcagcca
     3181 gaggatgcgg acaaccagcg gaacgtcacc cgcatgggta gtcagccccc ggacaccagc
     3241 accactgtgc acattcctgt gacgctgacc ggcccccccg gggagaccac ggtagtgccc
     3301 agtggaaacg tggacctgga aagtcaagca gaggggaaga aggaggtgga aaccagcgat
     3361 gtgatgagga gcggccctcg gcccattgtc ccgtacagtt ccatgttctg tttaagccca
     3421 accaacctgc tgcgccgctg ctgccactac attgtgacca tgtaactagt gcggccgcaa
     3481 ttccgggggg gggggggggg gggacagctc agggctgcga tttcgcgcca aacttgacgg
     3541 caatcctagc gtgaaggctg gtaggatttt atccccgctg ccatcatggt tcgaccattg
     3601 aactgcatcg tcgccgtgtc ccaaaatatg gggattggca agaacggaga cctaccctgg
     3661 cctccgctca ggaacgagtt caagtacttc caaagaatga ccacaacctc ttcagtggaa
     3721 ggtaaacaga atctggtgat tatgggtagg aaaacctggt tctccattcc tgagaagaat
     3781 cgacctttaa aggacagaat taatatagtt ctcagtagag aactcaaaga accaccacga
     3841 ggagctcatt ttcttgccaa aagtttggat gatgccttaa gacttattga acaaccggaa
     3901 ttggcaagta aagtagacat ggtttggata gtcggaggca gttctgttta ccaggaagcc
     3961 atgaatcaac caggccacct cagactcttt gtgacaagga tcatgcagga atttgaaagt
     4021 gacacgtttt tcccagaaat tgatttgggg aaatataaac ttctcccaga atacccaggc
     4081 gtcctctctg aggtccagga ggaaaaaggc atcaagtata agtttgaagt ctacgagaag
     4141 aaagactaac aggaagatgc tttcaagttc tctgctcccc tcctaaagct atgcattttt
     4201 ataagaccat gggacttttg ctggctttag atcataatca gccataccac atttgtagag
     4261 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     4321 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4381 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4441 ctcatcaatg tatcttatca tgtctggatc cccggccaac ggtctggtga cccggctgcg
     4501 agagctcggt gtacctgaga cgcgagtaag cccttgagtc aaagacgtag tcgttgcaag
     4561 tccgcaccag gtactgatat cccaccaaaa agtgcggcgg cggctggcgg tagaggggcc
     4621 agcgtagggt ggccggggct ccgggggcga ggtcttccaa cataaggcga tgatatccgt
     4681 agatgtacct ggacatccag gtgatgccgg cggcggtggt ggaggcgcgc ggaaagtcgc
     4741 ggacgcggtt ccagatgttg cgcagcggca aaaagtgctc catggtcggg acgctctggc
     4801 cggtgaggcg tgcgcagtcg ttgacgctct agaccgtgca aaaggagagc ctgtaagcgg
     4861 gcactcttcc gtggtctggt ggataaattc gcaagggtat catggcggac gaccggggtt
     4921 cgaaccccgg atccggccgt ccgccgtgat ccatgcggtt accgcccgcg tgtcgaaccc
     4981 aggtgtgcga cgtcagacaa cgggggagcg ctccttttgg cttccttcca ggcgcggcgg
     5041 ctgctgcgct agcttttttg gcgagctcga attaattctg cattaatgaa tcggccaacg
     5101 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5161 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5221 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5281 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5341 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5401 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5461 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5521 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5581 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5641 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5701 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5761 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5821 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5941 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6001 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6061 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6121 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6181 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6241 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6301 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6361 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6421 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6481 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6541 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6601 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6661 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6721 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6781 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6841 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6901 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6961 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7021 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     7081 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     7141 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     7201 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     7261 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     7321 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     7381 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     7441 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     7501 gtcacgacgt tgtaaaacga cggccagtgc caagcttttt gcaaaagcct aggcctccaa
     7561 aaaagcctcc tcactacttc tggaatagct cagaggccga ggcggcctcg gcctctgcat
     7621 aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc ggagttaggg
     7681 gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga tgcatgcttt
     7741 gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct gactaattga
     7801 gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca caccctaact
     7861 gacacacatt ccacaggatc cggtcgcgcg aatttcgagc ggtgttccgc ggtcctcctc
     7921 gtatagaaac tcggaccact ctgagacgaa ggctcgcgtc caggccagca cgaaggaggc
     7981 taagtgggag gggtagcggt cgttgtccac tagggggtcc actcgctcca gggtgtgaag
     8041 acacatgtcg ccctcttcgg catcaaggaa ggtgattggt ttataggtgt aggccacgtg
     8101 accgggtgtt cctgaagggg ggctataaaa gggggtgggg gcgcgttcgt cctcactctc
     8161 ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc ggttgaggac aaactcttcg
     8221 cggtctttcc agtactcttg gatcggaaac ccgtcggcct ccgaacggta ctccgccacc
     8281 gagggacctg agcgagtccg catcgaccgg atcggaaaac ctctcgactg ttggggtgag
     8341 tactccctct caaaagcggg catgacttct gcgctaagat tgtcagtttc caaaaacgag
     8401 gaggatttga tattcacctg gcccgcggtg atgcctttga gggtggccgc gtccatctgg
     8461 tcagaaaaga caatcttttt gttgtcaagc ttgaggtgtg gcaggcttga gatctggcca
     8521 tacacttgag tgacaatgac atccactttg cctttctctc cacaggtgtc cactcccagg
     8581 tccaactgca ggcgagcctg aattcctcga gggtaccggg ccccccctcg aggtcgaagc
     8641 ttgatc
//
