LOCUS       sequence-176915-        8906 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Alpha2 delta-1 V6.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8906)
  AUTHORS   Kadurin I, Ferron L, Rothwell SW, Meyer JO, Douglas LR, Bauer CS, 
            Lana B, Margas W, Alexopoulos O, Nieto-Rostro M, Pratt WS, Dolphin 
            AC
  TITLE     Proteolytic maturation of alpha2delta represents a checkpoint for 
            activation and neuronal trafficking of latent calcium channels.
  JOURNAL   Elife. 2016 Oct 26;5. pii: e21143. doi: 10.7554/eLife.21143.
  PUBMED    27782881
REFERENCE   2  (bases 1 to 8906)
  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: "Elife."; 
            date: "2016-10-26"; volume: "5. pii: e21143. doi"; pages: " 
            10.7554/eLife.21143"
COMMENT     Sequence Label: pMT2 Alpha2 delta-1 V6
FEATURES             Location/Qualifiers
     source          1..8906
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(2827..2844)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3818..4381
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    4568..4702
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4605..4624)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4659..4678
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5375..5392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5546..6134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5626..5645)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6305..7096,7097..7165))
                     /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     6928..6947
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7166..7270)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7338..7356
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7394..7416)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7516..7535
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7729..7751
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7743..7760
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7744..7760
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(7770..8099)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7784..7919
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8084..8104
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     intron          8568..8797
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     8780..8799
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 atggctgctg gctgcctgct ggccttgact ctgacacttt tccaatcttg gctgatcggc
       61 ccctcgagcg aggagccctt cccttcgccc gtcactatca agtcatgggt ggataaaatg
      121 caggaagacc ttgtcacact ggcaaagaca gcaagtggag tgactcagct tgctgacatt
      181 tatgaaaaat accaagattt gtatactgtg gagccaaata atgcacgcca actggttgaa
      241 attgcagcta gagacattga gaaacttctg agcaacagat ctaaagccct ggtgcgcctg
      301 gctatggaag cagagaaagt ccaagccgcc caccaatgga gggaagattt tgcaagcaac
      361 gaagttgtct actacaacgc taaagacgat cttgatcctg aaagaaatga gagtgagtca
      421 ggcagccaac ggattaaacc tgttttcatc gaagacgcta attttggacg ccagatatcc
      481 tatcaacatg cagcggtcca tattcccacg gacatctatg agggctcaac catagtgtta
      541 aacgaactca actggacaag tgccttagat gaagttttca aaagaaatcg agatgaagac
      601 cctacattgc tgtggcaagt gtttggcagc gccaccggcc tggcccgata ttatccagct
      661 tctccgtggg tggataacag cagaactcca aacaagatcg atctctacga tgtacgcaga
      721 agaccatggt acatccaggg agctgcatcc cccaaagaca tgctcattct ggtggacgtg
      781 agtggaagtg tgagcggatt gactctgaaa ctcatccgga catctgtctc tgagatgtta
      841 gaaacccttt ccgatgatga tttcgtgaat gtagcttcat ttaacagcaa tgctcaggat
      901 gtgagttgtt tccagcacct ggttcaagcg aatgtaagga acaagaaggt gctgaaggat
      961 gcggtgaata acatcacagc aaaggggata acagattaca agaaaggctt tagcttcgcg
     1021 tttgaacagc tacttaatta taatgtttcc agagccaatt gtaataagat catcatgtta
     1081 ttcacggatg gaggagaaga gagagcccag gagatatttg ccaagtacaa taaagacaaa
     1141 aaagtccgcg tgtttacatt ttctgttggt caacataatt atgacagagg acctattcag
     1201 tggatggctt gtgaaaacaa aggttattat tatgagattc cctctattgg tgcaataaga
     1261 atcaatactc aggaatatct agatgttctg ggaagaccaa tggttttagc cggggacaaa
     1321 gcaaagcaag tccagtggac aaatgtgtat ttggatgctc tggagctggg acttgtcatt
     1381 actggaactc taccggtctt caacgtcact ggccagtctg aaaataagac taacttgaag
     1441 aaccagttga ttcttggtgt gatgggcgtt gatgtgtctc tggaagatat taagagattg
     1501 acaccacgtt ttacactgtg ccccaacggt tactattttg caatcgatcc taatggttat
     1561 gtcttactgc atccaaatct tcagccaaag aaccccaaat ctcaggagcc ggtcacactg
     1621 gattttctcg atgctgagtt agagaatgat attaaagtgg agattcgaaa taaaatgata
     1681 gacggagaaa gtggagaaaa aacattcaga actctggtca aatctcaaga tgagagatat
     1741 attgacaaag gaaatcgaac atacacatgg acacctgtca atggcacaga ttacagtttg
     1801 gccttggtat tgccaaccta cagtttttac tatataaaag ccaaaataga agagacaata
     1861 actcaggcca gatcaaaaaa ggggaaaatg aaggattcag aaaccctgaa gccagacaat
     1921 tttgaagaat ctggctacac tttcatagca ccaagggaat actgcaatga cctgaaacct
     1981 tcagataata acactgaatt tcttttaaat ttcaatgaat ttattgatag gaaaactcca
     2041 aacaaccctt cctgtaatac agatttgatt aatagaatct tgctggatgc gggttttaca
     2101 aatgaacttg tccaaaatta ttggagtaaa cagaaaaata tcaaaggagt gaaggcacgc
     2161 tttgtggtga ctgatggcgg aattacaaga gtttacccca aagaggctgg agaaaactgg
     2221 caagagaacc cagagacgta tgaggacagc ttctacaaac ggagcctaga taacgataac
     2281 tacgttttca ctgcgcccta cttcaacaaa agtggacctg gtgcctatga atctggaatt
     2341 atggtaagca aagctgtaga actgtacatc caaggaaaac ttcttaagcc tgcagttgtg
     2401 ggaattaaaa ttgacgtaaa ctcctggata gaaaatttta ccaaaacttc aatcagggat
     2461 ccgtgtgctg gtccagtttg tgactgcaaa agaaacagtg atgtaatgga ctgtgtcatt
     2521 ctagatgatg gtggatttct tctgatggcc aatcatgatg attacactaa tcagattgga
     2581 cgtttttttg gagagattga cccgagcatg atgagacacc tggttaatat atcactttat
     2641 gcattcaata aatcatatga ctatcagtct gtgtgtgatc caggggcagc accaaagcaa
     2701 ggagcaggac atcgctcagc ctatgtgcca tcaattacag acatactcca gattggctgg
     2761 tgggccactg ctgccgcctg gtctattctc cagcagctac tcttgagttt gacatttcca
     2821 cggctcgtgg tggtggtggt ggtggaagag gatgacttca cagcttccct gtctaagcag
     2881 agctgcatca cagaacaaac tcagtacttc ttcaagaacg atactaaatc attcagtggt
     2941 ttactggact gtggaaactg ttccaggatc tttcatgttg agaagcttat gaacaccaac
     3001 ctagtgttca taatggtgga gagcaaggga acatgtccgt gtgacacgcg gctgctcatg
     3061 caagcagaac agacttctga tggtccagat ccttgcgaca tggttaagca gcccagatac
     3121 cgaaaaggac ctgatgtctg ctttgataac aatgtgctgg aggattatac cgactgtggt
     3181 ggtgtttctg gattaaaccc ttccttatgg tctatctttg gactccaatt tatactcctt
     3241 tggctggtat ctggcagcag acactatcta tggtgaccct ctaaacccat gatctgcaac
     3301 ttaattccat gccctgccaa aacatgagcc ctcagttaca ttaaaataga gcccgttgaa
     3361 gaagctccac agaaccttag ctgggcctct accctgatgt aactgtgagg cttagattcc
     3421 tgaccatcgc tggctgcatg tcagggtgtt ggaaactgaa actcgtgtga acatgctgca
     3481 tcatctgtgt ccaacatcaa agcaaagctc tactcattct ctgtgggagg tttgggagtt
     3541 cgctgttgca ctgttggaga ttacgtggaa aaggctcccc cacgcaggcc ttatgaaaag
     3601 aagcagtctt gactttgacc tagcatttca acctgctgca gttttaccaa gccgaattcc
     3661 tgcagcccgg gggatccact agatccatca cactggcggc cgcgtcgacg aattccgggg
     3721 gggggggggg gggggacagc tcagggctgc gatttcgcgc caaacttgac ggcaatccta
     3781 gcgtgaaggc tggtaggatt ttatccccgc tgccatcatg gttcgaccat tgaactgcat
     3841 cgtcgccgtg tcccaaaata tggggattgg caagaacgga gacctaccct ggcctccgct
     3901 caggaacgag ttcaagtact tccaaagaat gaccacaacc tcttcagtgg aaggtaaaca
     3961 gaatctggtg attatgggta ggaaaacctg gttctccatt cctgagaaga atcgaccttt
     4021 aaaggacaga attaatatag ttctcagtag agaactcaaa gaaccaccac gaggagctca
     4081 ttttcttgcc aaaagtttgg atgatgcctt aagacttatt gaacaaccgg aattggcaag
     4141 taaagtagac atggtttgga tagtcggagg cagttctgtt taccaggaag ccatgaatca
     4201 accaggccac ctcagactct ttgtgacaag gatcatgcag gaatttgaaa gtgacacgtt
     4261 tttcccagaa attgatttgg ggaaatataa acttctccca gaatacccag gcgtcctctc
     4321 tgaggtccag gaggaaaaag gcatcaagta taagtttgaa gtctacgaga agaaagacta
     4381 acaggaagat gctttcaagt tctctgctcc cctcctaaag ctatgcattt ttataagacc
     4441 atgggacttt tgctggcttt agatcataat cagccatacc acatttgtag aggttttact
     4501 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4561 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4621 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4681 tgtatcttat catgtctgga tccccggcca acggtctggt gacccggctg cgagagctcg
     4741 gtgtacctga gacgcgagta agcccttgag tcaaagacgt agtcgttgca agtccgcacc
     4801 aggtactgat atcccaccaa aaagtgcggc ggcggctggc ggtagagggg ccagcgtagg
     4861 gtggccgggg ctccgggggc gaggtcttcc aacataaggc gatgatatcc gtagatgtac
     4921 ctggacatcc aggtgatgcc ggcggcggtg gtggaggcgc gcggaaagtc gcggacgcgg
     4981 ttccagatgt tgcgcagcgg caaaaagtgc tccatggtcg ggacgctctg gccggtgagg
     5041 cgtgcgcagt cgttgacgct ctagaccgtg caaaaggaga gcctgtaagc gggcactctt
     5101 ccgtggtctg gtggataaat tcgcaagggt atcatggcgg acgaccgggg ttcgaacccc
     5161 ggatccggcc gtccgccgtg atccatgcgg ttaccgcccg cgtgtcgaac ccaggtgtgc
     5221 gacgtcagac aacgggggag cgctcctttt ggcttccttc caggcgcggc ggctgctgcg
     5281 ctagcttttt tggcgagctc gaattaattc tgcattaatg aatcggccaa cgcgcgggga
     5341 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5401 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5461 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5521 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5581 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5641 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5701 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5761 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5821 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5881 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5941 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6001 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6061 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6121 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6181 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6241 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6301 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6361 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6421 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6481 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6541 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6601 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6661 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6721 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6781 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6841 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6901 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6961 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7021 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7081 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7141 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7201 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7261 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     7321 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     7381 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     7441 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     7501 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     7561 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     7621 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     7681 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     7741 gttgtaaaac gacggccagt gccaagcttt ttgcaaaagc ctaggcctcc aaaaaagcct
     7801 cctcactact tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa
     7861 aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg
     7921 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
     7981 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
     8041 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
     8101 ttccacagga tccggtcgcg cgaatttcga gcggtgttcc gcggtcctcc tcgtatagaa
     8161 actcggacca ctctgagacg aaggctcgcg tccaggccag cacgaaggag gctaagtggg
     8221 aggggtagcg gtcgttgtcc actagggggt ccactcgctc cagggtgtga agacacatgt
     8281 cgccctcttc ggcatcaagg aaggtgattg gtttataggt gtaggccacg tgaccgggtg
     8341 ttcctgaagg ggggctataa aagggggtgg gggcgcgttc gtcctcactc tcttccgcat
     8401 cgctgtctgc gagggccagc tgttgggctc gcggttgagg acaaactctt cgcggtcttt
     8461 ccagtactct tggatcggaa acccgtcggc ctccgaacgg tactccgcca ccgagggacc
     8521 tgagcgagtc cgcatcgacc ggatcggaaa acctctcgac tgttggggtg agtactccct
     8581 ctcaaaagcg ggcatgactt ctgcgctaag attgtcagtt tccaaaaacg aggaggattt
     8641 gatattcacc tggcccgcgg tgatgccttt gagggtggcc gcgtccatct ggtcagaaaa
     8701 gacaatcttt ttgttgtcaa gcttgaggtg tggcaggctt gagatctggc catacacttg
     8761 agtgacaatg acatccactt tgcctttctc tccacaggtg tccactccca ggtccaactg
     8821 caggcgagcc tgaattcctc gagggtaccg agctcggatc cactagtaac ggccgccagt
     8881 gtgctggaat tctgcagatc gcgaag
//
