LOCUS       sequence-413000-        8531 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of rat alpha2delta-1 calcium channel auxillary subunit 
            with a single HA tag inserted after aa549.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8531)
  AUTHORS   Kadurin I, Alvarez-Laviada A, Ng SF, Walker-Gray R, D'Arco M, Fadel 
            MG, Pratt WS, Dolphin AC
  TITLE     Calcium currents are enhanced by alpha2delta-1 lacking its membrane 
            anchor.
  JOURNAL   J Biol Chem. 2012 Sep 28;287(40):33554-66. Epub 2012 Aug 6.
  PUBMED    22869375
REFERENCE   2  (bases 1 to 8531)
  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 Biol 
            Chem."; date: "2012-09-28"; volume: "287(40)"; pages: "33554-66. 
            Epub 2012 Aug 6"
COMMENT     Sequence Label: cacna2d1 HA pCAGGS
FEATURES             Location/Qualifiers
     source          1..8531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        385..660
                     /label=chicken beta-actin promoter
     intron          661..1677
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1601..1623
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1685..1704
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3393..3419
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(5552..5571)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5617..5672
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5671..5690)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6033..6049)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6033..6049)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6046..6068)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6057..6073
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6081..6087,6088..6105,6106..6111))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6126..6147
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6206..6401
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6252..6387
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6407..6541
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6444..6463)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6498..6517
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6609..6626)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6780..7368)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6860..6879)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7539..8330,8331..8399))
                     /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     8162..8181
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8400..8504)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtcgagg tgagccccac gttctgcttc actctcccca
      421 tctccccccc ctccccaccc ccaattttgt atttatttat tttttaatta ttttgtgcag
      481 cgatgggggc gggggggggg ggggggggcg ccaggggggg gggggggggg ggaggggggg
      541 gggggggsga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt
      601 ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg
      661 ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc gcgccgcccg
      721 ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg cccttctcct
      781 ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg gctgcgtgaa
      841 agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg gggtgcgtgc
      901 gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg ctgtgagcgc
      961 tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag cgcggccggg
     1021 ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc ccccctgcac
     1141 ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg tacggggcgt
     1201 ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg kgggggtgcc gggcggggcg
     1261 gggccgcctc gggccggggr gggctcgggg gaggggcgcg gcggcccccg gagcgccggc
     1321 ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg cgagagggcg
     1381 cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg ccgccgcacc
     1441 ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat gggcggggag
     1501 ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg gggctgtccg
     1561 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     1621 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     1681 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt ctgcagatcg
     1741 cgaagatggc tgctggctgc ctgctggcct tgactctgac acttttccaa tcttggctga
     1801 tcggcccctc gagcgaggag cccttccctt cgcccgtcac tatcaagtca tgggtggata
     1861 aaatgcagga agaccttgtc acactggcaa agacagcaag tggagtgact cagcttgctg
     1921 acatttatga aaaataccaa gatttgtata ctgtggagcc aaataatgca cgccaactgg
     1981 ttgaaattgc agctagagac attgagaaac ttctgagcaa cagatctaaa gccctggtgc
     2041 gcctggctat ggaagcagag aaagtccaag ccgcccacca atggagggaa gattttgcaa
     2101 gcaacgaagt tgtctactac aacgctaaag acgatcttga tcctgaaaga aatgagagtg
     2161 agtcaggcag ccaacggatt aaacctgttt tcatcgaaga cgctaatttt ggacgccaga
     2221 tatcctatca acatgcagcg gtccatattc ccacggacat ctatgagggc tcaaccatag
     2281 tgttaaacga actcaactgg acaagtgcct tagatgaagt tttcaaaaga aatcgagatg
     2341 aagaccctac attgctgtgg caagtgtttg gcagcgccac cggcctggcc cgatattatc
     2401 cagcttctcc gtgggtggat aacagcagaa ctccaaacaa gatcgatctc tacgatgtac
     2461 gcagaagacc atggtacatc cagggagctg catcccccaa agacatgctc attctggtgg
     2521 acgtgagtgg aagtgtgagc ggattgactc tgaaactcat ccggacatct gtctctgaga
     2581 tgttagaaac cctttccgat gatgatttcg tgaatgtagc ttcatttaac agcaatgctc
     2641 aggatgtgag ttgtttccag cacctggttc aagcgaatgt aaggaacaag aaggtgctga
     2701 aggatgcggt gaataacatc acagcaaagg ggataacaga ttacaagaaa ggctttagct
     2761 tcgcgtttga acagctactt aattataatg tttccagagc caattgtaat aagatcatca
     2821 tgttattcac ggatggagga gaagagagag cccaggagat atttgccaag tacaataaag
     2881 acaaaaaagt ccgcgtgttt acattttctg ttggtcaaca taattatgac agaggaccta
     2941 ttcagtggat ggcttgtgaa aacaaaggtt attattatga gattccctct attggtgcaa
     3001 taagaatcaa tactcaggaa tatctagatg ttctgggaag accaatggtt ttagccgggg
     3061 acaaagcaaa gcaagtccag tggacaaatg tgtatttgga tgctctggag ctgggacttg
     3121 tcattactgg aactctaccg gtcttcaacg tcactggcca gtctgaaaat aagactaact
     3181 tgaagaacca gttgattctt ggtgtgatgg gcgttgatgt gtctctggaa gatattaaga
     3241 gattgacacc acgttttaca ctgtgcccca acggttacta ttttgcaatc gatcctaatg
     3301 gttatgtctt actgcatcca aatcttcagc caaagaaccc caaatctcag gagccggtca
     3361 cactggattt tctcgatgct gagttagaga attaccctta cgacgtccct gactacgccg
     3421 atattaaagt ggagattcga aataaaatga tagacggaga aagtggagaa aaaacattca
     3481 gaactctggt caaatctcaa gatgagagat atattgacaa aggaaatcga acatacacat
     3541 ggacacctgt caatggcaca gattacagtt tggccttggt attgccaacc tacagttttt
     3601 actatataaa agccaaaata gaagagacaa taactcaggc cagatcaaaa aaggggaaaa
     3661 tgaaggattc agaaaccctg aagccagaca attttgaaga atctggctac actttcatag
     3721 caccaaggga atactgcaat gacctgaaac cttcagataa taacactgaa tttcttttaa
     3781 atttcaatga atttattgat aggaaaactc caaacaaccc ttcctgtaat acagatttga
     3841 ttaatagaat cttgctggat gcgggtttta caaatgaact tgtccaaaat tattggagta
     3901 aacagaaaaa tatcaaagga gtgaaggcac gctttgtggt gactgatggc ggaattacaa
     3961 gagtttaccc caaagaggct ggagaaaact ggcaagagaa cccagagacg tatgaggaca
     4021 gcttctacaa acggagccta gataacgata actacgtttt cactgcgccc tacttcaaca
     4081 aaagtggacc tggtgcctat gaatctggaa ttatggtaag caaagctgta gaactgtaca
     4141 tccaaggaaa acttcttaag cctgcagttg tgggaattaa aattgacgta aactcctgga
     4201 tagaaaattt taccaaaact tcaatcaggg atccgtgtgc tggtccagtt tgtgactgca
     4261 aaagaaacag tgatgtaatg gactgtgtca ttctagatga tggtggattt cttctgatgg
     4321 ccaatcatga tgattacact aatcagattg gacgtttttt tggagagatt gacccgagca
     4381 tgatgagaca cctggttaat atatcacttt atgcattcaa taaatcatat gactatcagt
     4441 ctgtgtgtga tccaggggca gcaccaaagc aaggagcagg acatcgctca gcctatgtgc
     4501 catcaattac agacatactc cagattggct ggtgggccac tgctgccgcc tggtctattc
     4561 tccagcagct actcttgagt ttgacatttc cacggctcct tgaggcagtt gaaatggaag
     4621 aggatgactt cacagcttcc ctgtctaagc agagctgcat cacagaacaa actcagtact
     4681 tcttcaagaa cgatactaaa tcattcagtg gtttactgga ctgtggaaac tgttccagga
     4741 tctttcatgt tgagaagctt atgaacacca acctagtgtt cataatggtg gagagcaagg
     4801 gaacatgtcc gtgtgacacg cggctgctca tgcaagcaga acagacttct gatggtccag
     4861 atccttgcga catggttaag cagcccagat accgaaaagg acctgatgtc tgctttgata
     4921 acaatgtgct ggaggattat accgactgtg gtggtgtttc tggattaaac ccttccttat
     4981 ggtctatctt tggactccaa tttatactcc tttggctggt atctggcagc agacactatc
     5041 tatggtgacc ctctaaaccc atgatctgca acttaattcc atgccctgcc aaaacatgag
     5101 ccctcagtta cattaaaata gagcccgttg aagaagctcc acagaacctt agctgggcct
     5161 ctaccctgat gtaactgtga ggcttagatt cctgaccatc gctggctgca tgtcagggtg
     5221 ttggaaactg aaactcgtgt gaacatgctg catcatctgt gtccaacatc aaagcaaagc
     5281 tctactcatt ctctgtggga ggtttgggag ttcgctgttg cactgttgga gattacgtgg
     5341 aaaaggctcc cccacgcagg ccttatgaaa agaagcagtc ttgactttga cctagcattt
     5401 caacctgctg cagttttacc aagccgaatt cgcatgcggc cgctagctcg aggtaccgat
     5461 atcgagctca attcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg
     5521 ccaatgccct ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg
     5581 acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt attttcattg
     5641 caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc
     5701 atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg
     5761 ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg
     5821 tccattcctt attccataga aaagccttga cttgaggtta gatttttttt atattttgtt
     5881 ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat
     5941 ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc
     6001 tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     6061 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     6121 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     6181 ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc
     6241 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     6301 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     6361 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact tgtttattgc
     6421 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6481 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     6541 ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6601 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6661 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6721 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6781 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6841 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6901 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6961 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7021 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7081 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7141 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7201 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7261 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7321 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7381 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7441 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7501 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7561 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7621 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7681 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7741 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7801 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7861 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7921 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7981 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8041 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8101 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8161 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8221 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8281 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8341 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8401 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8461 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8521 gtgccacctg g
//
