LOCUS       sequence-90810-e        8505 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of HA tagged mouse Alpha2 delta-3 in mamalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8505)
  AUTHORS   Davies A, Kadurin I, Alvarez-Laviada A, Douglas L, Nieto-Rostro M, 
            Bauer CS, Pratt WS, Dolphin AC
  TITLE     The alpha2delta subunits of voltage-gated calcium channels form 
            GPI-anchored proteins, a posttranslational modification essential 
            for function.
  JOURNAL   Proc Natl Acad Sci U S A. 2010 Jan 26;107(4):1654-9. doi: 
            10.1073/pnas.0908735107. Epub 2010 Jan 4.
  PUBMED    20080692
REFERENCE   2  (bases 1 to 8505)
  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.0908735107"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2010-01-26- 26"; volume: "107"; issue: "4"; pages: "1654-9"
COMMENT     Sequence Label: Mouse Alpha2 delta-3 HA pMT2
FEATURES             Location/Qualifiers
     source          1..8505
                     /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"
     CDS             2898..2924
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4553..5116
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     polyA_signal    5304..5438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5341..5360)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5395..5414
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6111..6128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6282..6870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6362..6381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7041..7832,7833..7901))
                     /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     7664..7683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7902..8006)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8074..8092
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8130..8152)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8252..8271
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8465..8487
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8479..8496
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8480..8496
                     /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 attgcggccg
     1081 ctccagtcgc cgccgccgcc gccgccgcca ggatggccgg gccgggctcg ctgtgctgcg
     1141 cgtcccgggg ggcctcggcg ctcctagcca ccgcgcttct ctacgccgct ctgggggacg
     1201 tggtgcgctc cgagcagcag atcccgctct ccgtagtgaa gctgtgggcc tccgcttttg
     1261 gtggggagat aaaatccatc gctgccaagt actcgggctc ccagcttctg caaaagaaat
     1321 acaaagagta tgagaaggac gttgccattg aagaaatcga cggtctccaa ctggtgaaga
     1381 agttggctaa aatcatggag gagatgttcc acaagaagtc cgaggcagtg aggcgtcttg
     1441 tggaggctgc agaggaggca cacctgaaac atgaatttga tgctgacttg cagtatgaat
     1501 acttcaatgc tgtgctgatc aacgagaggg acaaagacgg gaactttttg gaattgggaa
     1561 aggaattcat cttagccccc aatgaccatt ttaataattt gcctgtgaac atcagtctga
     1621 gtgatgtgca agtgccaacg aacatgtaca acaaagatcc tgccattgtc aatggagtgt
     1681 attggtctga atctctaaac aaagtttttg tggataactt tgatcgggat ccgtctctca
     1741 tatggcagta ctttggaagt gcaaagggct ttttcagaca gtacccaggg attaaatggg
     1801 aaccagatga gaatggagtc attgcctttg actgcaggaa cagaaaatgg tacatccagg
     1861 cagcgacttc tccaaaggat gttgtcattt tggtggacgt cagtgggagc atgaaaggac
     1921 tccgcttgac catcgccaag caaacagtgt cctcaatact ggatactctg ggtgatgatg
     1981 acttcttcaa catcatcacg tataacgaag agcttcacta tgtggaacct tgtctgaacg
     2041 gaacactggt tcaagctgac aggaccaaca aggagcactt cagggagcat ttggacaaac
     2101 tttttgccaa agggattgga atgctggata tcgcactgaa cgaggccttc aacatactga
     2161 gcgatttcaa ccacactgga caaggaagca tttgcagcca ggccatcatg ctcataactg
     2221 atggggcagt ggacacctat gataccatct ttgcaaaata taattggcca gaccgaaagg
     2281 ttcgaatctt tacttacctc attggacggg aggctgcttt tgccgacaat ctcaagtgga
     2341 tggcttgtgc taacaaagga tttttcaccc agatctccac cttggctgat gtgcaggaaa
     2401 atgtcatgga atacctccat gtgcttagcc gacctaaagt cattgaccag gagcatgatg
     2461 tggtgtggac cgaagcgtac attgacagca ccctccctca ggctcaaaag cttgctgatg
     2521 atcagggcct cgtcttgatg accacggtgg ccatgcctgt gtttagtaag cagaacgaaa
     2581 ctaggtcaaa gggcattctt ctgggtgtgg ttggcacaga tgtcccagtt aaagagcttc
     2641 tgaagaccat ccccaaatac aagttaggaa ttcatggtta cgccttcgcc atcacgaata
     2701 atggatatat cctgacgcac ccggagctca ggccactgta tgaagaagga aaaaaacgga
     2761 ggaaacccaa ctacagtagt gtggatctct ctgaagtcga gtgggaagac cgggatgatg
     2821 tgttacgaaa tgccatggta aatcggaaga ctgggaaatt ctccatggaa gtgaagaaga
     2881 ccgtggacaa agggaaatac ccttacgacg tccctgacta cgcccgggtt ttggtgatga
     2941 ccaatgacta ctactataca gacatcaagg gtactccttt cagtttaggt gtggcgctct
     3001 ccaggggcca tgggaaatac ttcttccgag ggaatgtaac cattgaagaa ggcctccatg
     3061 acttagaaca tcctgacgtg tccttggcag atgaatggtc ctactgcaac actgacctgc
     3121 acccagagca ccgccatcta tctcaactgg aagccattaa gctctacctc aaaggcaagg
     3181 agcctctgct tcaatgtgac aaagaattga ttcaagaagt cctttttgat gctgtggtga
     3241 gcgcccctat tgaagcctat tggacgagcc tggccctcaa caaatctgag aattctgaca
     3301 agggtgtaga ggtcgccttc ctcggcactc gcacaggcct ctcaagaatc aacctgtttg
     3361 tgggggccga acaactcacc aatcaggact ttctgaaggc tggagacaaa gagaacattt
     3421 ttaatgccga tcatttccct ctctggtaca gaagagctgc cgagcagatc gcaggaagct
     3481 ttgtctattc catccccttc agcacaggaa cagtcaacaa aagcaatgtg gtgacagcaa
     3541 gtacctccat ccagctcctg gatgagcgga aatctcccgt ggtggcagct gtaggcattc
     3601 agatgaaact tgaattcttc caaaggaagt tctggactgc cagcagacag tgtgcctccc
     3661 tggatggcaa atgctccata agctgcgatg acgagactgt gaactgttac cttatagaca
     3721 ataacggatt cattctggtg tctgaagact acacacagac tggagatttt tttggtgagg
     3781 tggaaggagc tgtcatgaac aagttgttaa caatgggttc ctttaaaaga ataaccttgt
     3841 acgactacca agccatgtgt agagccaaca aggagagcag tgacagtgcc catggacttc
     3901 tggaccccta taaggccttt ctctctgcag ccaagtggat aatgacggaa cttgtcttgt
     3961 tcctggtgga gtttaacctg tgcagttggt ggcactccga catgacagct aaagcccaga
     4021 aactgaaaca gaccctggaa ccttgtgata ctgaataccc agcctttgtt tctgaacgca
     4081 ccatcaagga gaccacaggg aacattgctt gcgaagactg ctccaagtcc ttcgtcatcc
     4141 agcaaatccc gagtagcaat ctgttcatgg tggtggtgga cagtagctgt ctctgtgagt
     4201 ccgtggctcc tataaccatg gcacccattg aaatcaggta taatgaatcc cttaagtgtg
     4261 aacggttaaa ggctcagaag atcagacgac gtccagaatc ctgccacggc ttccatcctg
     4321 aggagaatgc aagagagtgt gggggtgcat cgagtcttca ggcccaggcg gccttgctgc
     4381 tgctgcccct ggtttcgagt ctcttctcaa ggtgagcggc cgcactagtc ccgggtacct
     4441 cgaggaattc cggggggggg gggggggggg acagctcagg gctgcgattt cgcgccaaac
     4501 ttgacggcaa tcctagcgtg aaggctggta ggattttatc cccgctgcca tcatggttcg
     4561 accattgaac tgcatcgtcg ccgtgtccca aaatatgggg attggcaaga acggagacct
     4621 accctggcct ccgctcagga acgagttcaa gtacttccaa agaatgacca caacctcttc
     4681 agtggaaggt aaacagaatc tggtgattat gggtaggaaa acctggttct ccattcctga
     4741 gaagaatcga cctttaaagg acagaattaa tatagttctc agtagagaac tcaaagaacc
     4801 accacgagga gctcattttc ttgccaaaag tttggatgat gccttaagac ttattgaaca
     4861 accggaattg gcaagtaaag tagacatggt ttggatagtc ggaggcagtt ctgtttacca
     4921 ggaagccatg aatcaaccag gccacctcag actctttgtg acaaggatca tgcaggaatt
     4981 tgaaagtgac acgtttttcc cagaaattga tttggggaaa tataaacttc tcccagaata
     5041 cccaggcgtc ctctctgagg tccaggagga aaaaggcatc aagtataagt ttgaagtcta
     5101 cgagaagaaa gactaacagg aagatgcttt caagttctct gctcccctcc taaagctatg
     5161 cattttttat aagaccatgg gacttttgct ggctttagat cataatcagc cataccacat
     5221 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     5281 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     5341 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     5401 tgtccaaact catcaatgta tcttatcatg tctggatccc cggccaacgg tctggtgacc
     5461 cggctgcgag agctcggtgt acctgagacg cgagtaagcc cttgagtcaa agacgtagtc
     5521 gttgcaagtc cgcaccaggt actgatatcc caccaaaaag tgcggcggcg gctggcggta
     5581 gaggggccag cgtagggtgg ccggggctcc gggggcgagg tcttccaaca taaggcgatg
     5641 atatccgtag atgtacctgg acatccaggt gatgccggcg gcggtggtgg aggcgcgcgg
     5701 aaagtcgcgg acgcggttcc agatgttgcg cagcggcaaa aagtgctcca tggtcgggac
     5761 gctctggccg gtgaggcgtg cgcagtcgtt gacgctctag accgtgcaaa aggagagcct
     5821 gtaagcgggc actcttccgt ggtctggtgg ataaattcgc aagggtatca tggcggacga
     5881 ccggggttcg aaccccggat ccggccgtcc gccgtgatcc atccggttac cgcccgcgtg
     5941 tcgaacccag gtgtgcgacg tcagacaacg ggggagcgct ccttttggct tccttccagg
     6001 cgcggcggct gctgcgctag cttttttggc gagctcgaat taattctgca ttaatgaatc
     6061 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6121 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6181 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6241 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6301 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6361 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6421 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     6481 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6541 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6601 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6661 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6721 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6781 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6841 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6901 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6961 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7021 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7081 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7141 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7201 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7261 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7321 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7381 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7441 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7501 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7561 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7621 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7681 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7741 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7801 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7861 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7921 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7981 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     8041 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     8101 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     8161 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     8221 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     8281 caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     8341 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     8401 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     8461 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agctt
//
