LOCUS       sequence-415914-        9259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of the N-terminus, Domain I, Domain II and the II-III 
            loop (amino acids 1-1154) of rabbit Cav2.2 calcium channel with a 
            N-terminal GFP tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9259)
  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 9259)
  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: GFP Cav2.2 DomI + DomII pMT2
FEATURES             Location/Qualifiers
     source          1..9259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..29
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(248..352)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     420..438
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(476..498)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     598..617
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     811..833
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     825..842
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     826..842
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(852..1181)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      866..1001
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1166..1186
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          1650..1879
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     1862..1881
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             1949..2662
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1949..2662
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1979..2007)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2602..2622
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             6159..6722
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    6909..7043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6946..6965)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7000..7019
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7716..7733)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7887..8475)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7967..7986)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8646..9259,1..178,179..247))
                     /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"
ORIGIN
        1 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
       61 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
      121 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
      181 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
      241 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
      301 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
      361 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
      421 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
      481 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
      541 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
      601 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
      661 taaggagaaa ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag
      721 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      781 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      841 gtgccaagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
      901 agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg
      961 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     1021 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     1081 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     1141 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag gatccggtcg
     1201 cgcgaatttc gagcggtgtt ccgcggtcct cctcgtatag aaactcggac cactctgaga
     1261 cgaaggctcg cgtccaggcc agcacgaagg aggctaagtg ggaggggtag cggtcgttgt
     1321 ccactagggg gtccactcgc tccagggtgt gaagacacat gtcgccctct tcggcatcaa
     1381 ggaaggtgat tggtttatag gtgtaggcca cgtgaccggg tgttcctgaa ggggggctat
     1441 aaaagggggt gggggcgcgt tcgtcctcac tctcttccgc atcgctgtct gcgagggcca
     1501 gctgttgggc tcgcggttga ggacaaactc ttcgcggtct ttccagtact cttggatcgg
     1561 aaacccgtcg gcctccgaac ggtactccgc caccgaggga cctgagcgag tccgcatcga
     1621 ccggatcgga aaacctctcg actgttgggg tgagtactcc ctctcaaaag cgggcatgac
     1681 ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca cctggcccgc
     1741 ggtgatgcct ttgagggtgg ccgcgtccat ctggtcagaa aagacaatct ttttgttgtc
     1801 aagcttgagg tgtggcaggc ttgagatctg gccatacact tgagtgacaa tgacatccac
     1861 tttgcctttc tctccacagg tgtccactcc caggtccaac tgcaggcgag cctgaattcc
     1921 tcgagggtac cgagctcgga tctaaaaaat gagtaaagga gaagaacttt tcactggagt
     1981 tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg
     2041 agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg
     2101 aaaactacct gttccatggc caacacttgt cactactttc ggttatggtg ttcaatgctt
     2161 tgcgagatac ccagatcata tgaaacagca tgactttttc aagagtgcca tgcccgaagg
     2221 ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga cacgtgctga
     2281 agtcaagttt gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa
     2341 agaagatgga aacattcttg gacacaaatt ggaatacaac tataactcac acaatgtata
     2401 catcatggca gacaaacaaa agaatggaat caaagttaac ttcaaaatta gacacaacat
     2461 tgaagatgga agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg
     2521 ccctgtcctt ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc
     2581 caacgaaaag agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca
     2641 tggcatggat gaactataca aaatggtccg cttcggggac gagctgggcg gccgctatgg
     2701 gggcgcgggc ggcgcggagc gggcccgggg cggcggggcc ggcggggctg gcggcccggg
     2761 ccccgggggg ctgccgccgg gccagcgggt cctctacaaa cagtcgatcg cgcagcgcgc
     2821 gcggaccatg gcgctgtaca accccatccc ggtcaagcag aactgcttca ccgtcaaccg
     2881 ctcgctcttc gtcttcagcg aggacaacgt cgtccgcaaa tacgccaagc gcatcaccga
     2941 gtggcctccg ttcgagtaca tgatcctggc caccatcatc gccaactgca tcgtgctggc
     3001 cctggagcag cacctccctg acggggacaa gacgcctatg tccgagcgcc tggatgacac
     3061 cgagccctac ttcattggga tcttctgctt cgaggcgggg atcaagatcc tggccctggg
     3121 ctttgttctg cacaagggct cctatctgcg gaacggctgg aacgtcatgg acttcgtggt
     3181 agtcctcacg gggattcttg ccacagccgg aaccgacttc gacctgcgaa cgctgagggc
     3241 tgtgcgtgtg ctaaggcccc tgaagctggt gtctgggatt ccgagcttgc aggtggtgct
     3301 caagtccatc atgaaggcca tggtcccgct gctgcagatc gggctgctgc tcttcttcgc
     3361 catcctcatg tttgccatca tcggcctcga gttctacatg ggcaagttcc acaaggcctg
     3421 cttccccaac agcacagacc cagatcccgt gggtgacttc ccctgtggca aggaggcccc
     3481 ggcccggctc tgtgagggag acacggagtg ccgggagtac tgggccggac ccaactttgg
     3541 tatcaccaac tttgacaaca tcctgtttgc catcttgacc gtgttccagt gcatcaccat
     3601 ggagggctgg actgacatcc tctacaatac aaacgatgca gccggcaaca cgtggaactg
     3661 gctctacttc atccctctca tcatcatcgg ctccttcttc atgctcaacc tggtgctggg
     3721 tgtgctctca ggagagtttg ctaaggagcg cgagagagtg gagaaccgcc gcgccttcct
     3781 gaagctccgg cggcagcagc agattgagcg ggagctcaac gggtacctgg agtggatctt
     3841 caaggctgag gaagtcatgc tggctgagga ggacaggaac gcagaggaga agtcccctct
     3901 ggatgcagtg ttgaagagag cagctgccaa gaagagcaga agcgacctga tccaggccga
     3961 ggagggggag ggccggctca caggcctctg tgctcccggt tccccgttcg cccgtgccag
     4021 cctgaaaagc gggaagacgg agagctcgtc gtacttccgg aggaaggaga agatgttccg
     4081 cttcttcatc cggcgcatgg tgaaggcaca gagcttctac tggacggtgc tgtgtgtggt
     4141 ggcactgaac acactgtgcg tggccatggt gcactacaac cagccgcagc ggctcaccac
     4201 ggcattgtac tttgcagagt ttgttttcct gggtctcttc ctcacagaga tgtccctgaa
     4261 gatgtatggc ctggggccca ggagctactt ccggtcctcc ttcaactgtt ttgactttgg
     4321 ggtcatcgtg ggcagtatct ttgaagtggt ctgggctgct gtcaagccag ggacctcctt
     4381 cgggatcagt gttctgcggg ctctgcgctt actgagaatc ttcaaggtca ccaaatactg
     4441 gaactcccta cggaacctgg tggtgtctct gctcaactcc atgaagtcca tcatcagcct
     4501 cctcttcctg ctcttcctgt tcattgttgt ctttgctctg ctggggatgc agctgtttgg
     4561 aggccagttc aactttaaag acgagacccc aacgaccaac tttgacacct tccctgctgc
     4621 tatcctcacc gtcttccaga tcctgaccgg agaggactgg aatgcggtca tgtaccacgg
     4681 gatcgagtcg caaggtggcg tcagcagagg aatgttctcc tccttctact tcattgtcct
     4741 cactctgttt gggaactaca ccctgctgaa tgtcttcctg gccattgctg tggacaacct
     4801 tgccaatgct caggagctga ccaaggatga agaggagatg gaagaagcag ccaatcagaa
     4861 actcgccttg cagaaggcca aggaagtggc tgaagtcagc cccatgtctg ctgcgaacat
     4921 ctccattgcc gccaggcagc agaactcggc caaagcgcgc tcggtgtggg agcagcgagc
     4981 cagccagctg cggctgcaga acctgagggc cagctgcgag gcgctgtaca gcgagatgga
     5041 ccccgaggag aggctgcgct acgccaccgc gcgccacctg cggcccgacg tgaagacgca
     5101 cctggaccgg ccgctggtgg ttgagcccgg ccgcgacgct ccgcggggcc ccccgggggg
     5161 gaagtcccgg ccggatggct cggaggcccc cgagggtgcg gatcccccgc gcaggcatca
     5221 ccggcaccga gacaaggaca aggctcccgc cacggtcccc tcggcggggg aacaggacag
     5281 agccgaagct ctgagggcag agggcgggga gctggggccc cgcgaggagc gggggcggcc
     5341 acgccgcagc cgcagcaagg aggccccggg cgccccggag gtgcgcagcg atcgcggccg
     5401 tggcccgtgt cctgaaggtg gtcggcgcca ccaccggcgc gggtcaccgg aggaggcagc
     5461 tgagagggag ccccgccgcc accgcgcgca ccggcacggg cccgacccgg gcaaggaggg
     5521 cccagcgtcc gggaccaggg gcgagcggcg cgcacgacac cgcacgggcc cccgggcgtg
     5581 cccccgggag gctgagagca gcgaggagcc tgcgcggcgg caccgggctc ggcacaaggc
     5641 gccgcccacc caggagaccg cggagaagga caaggaggct gccgagaagg ggggcgaggc
     5701 cacggaggcc gagaaggaca aggaggctcg gaaccaccag cccaaggagc tgccctgtga
     5761 cctggaggcc atcggcatgc tgggagtggg cgctgtgcat acgctgccca gcacctgtct
     5821 gcagaaggtg gaggagcagc cagaggatgc ggacaaccag cggaacgtca cccgcatggg
     5881 tagtcagccc ccggacacca gcaccactgt gcacattcct gtgacgctga ccggcccccc
     5941 cggggagacc acggtagtgc ccagtggaaa cgtggacctg gaaagtcaag cagaggggaa
     6001 gaaggaggtg gaaaccagcg atgtgatgag gagcggccct cggcccattg tcccgtacag
     6061 ttccatgttc tgtttaagcc caaccaacct gctgcgccgc tgctgccact acattrtsmy
     6121 mrygtramkr stggtaggat tttatccccg ctgccatcat ggttcgacca ttgaactgca
     6181 tcgtcgccgt gtcccaaaat atggggattg gcaagaacgg agacctaccc tggcctccgc
     6241 tcaggaacga gttcaagtac ttccaaagaa tgaccacaac ctcttcagtg gaaggtaaac
     6301 agaatctggt gattatgggt aggaaaacct ggttctccat tcctgagaag aatcgacctt
     6361 taaaggacag aattaatata gttctcagta gagaactcaa agaaccacca cgaggagctc
     6421 attttcttgc caaaagtttg gatgatgcct taagacttat tgaacaaccg gaattggcaa
     6481 gtaaagtaga catggtttgg atagtcggag gcagttctgt ttaccaggaa gccatgaatc
     6541 aaccaggcca cctcagactc tttgtgacaa ggatcatgca ggaatttgaa agtgacacgt
     6601 ttttcccaga aattgatttg gggaaatata aacttctccc agaataccca ggcgtcctct
     6661 ctgaggtcca ggaggaaaaa ggcatcaagt ataagtttga agtctacgag aagaaagact
     6721 aacaggaaga tgctttcaag ttctctgctc ccctcctaaa gctatgcatt tttataagac
     6781 catgggactt ttgctggctt tagatcataa tcagccatac cacatttgta gaggttttac
     6841 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     6901 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     6961 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     7021 atgtatctta tcatgtctgg atccccggcc aacggtctgg tgacccggct gcgagagctc
     7081 ggtgtacctg agacgcgagt aagcccttga gtcaaagacg tagtcgttgc aagtccgcac
     7141 caggtactga tatcccacca aaaagtgcgg cggcggctgg cggtagaggg gccagcgtag
     7201 ggtggccggg gctccggggg cgaggtcttc caacataagg cgatgatatc cgtagatgta
     7261 cctggacatc caggtgatgc cggcggcggt ggtggaggcg cgcggaaagt cgcggacgcg
     7321 gttccagatg ttgcgcagcg gcaaaaagtg ctccatggtc gggacgctct ggccggtgag
     7381 gcgtgcgcag tcgttgacgc tctagaccgt gcaaaaggag agcctgtaag cgggcactct
     7441 tccgtggtct ggtggataaa ttcgcaaggg tatcatggcg gacgaccggg gttcgaaccc
     7501 cggatccggc cgtccgccgt gatccatgcg gttaccgccc gcgtgtcgaa cccaggtgtg
     7561 cgacgtcaga caacggggga gcgctccttt tggcttcctt ccaggcgcgg cggctgctgc
     7621 gctagctttt ttggcgagct cgaattaatt ctgcattaat gaatcggcca acgcgcgggg
     7681 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7741 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7801 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7861 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7921 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7981 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     8041 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     8101 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     8161 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     8221 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     8281 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     8341 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     8401 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     8461 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     8521 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     8581 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     8641 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     8701 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8761 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8821 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8881 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8941 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     9001 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     9061 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     9121 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     9181 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     9241 agttgctctt gcccggcgt
//
