LOCUS       sequence-415385-        9134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of rat alpha2delta-1 calcium channel auxillary subunit.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9134)
  TITLE     Dolphin Lab Plasmids 2023
REFERENCE   2  (bases 1 to 9134)
  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"
COMMENT     Sequence Label: cacna2d1 pcDNA3
FEATURES             Location/Qualifiers
     source          1..9134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      168..303
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     230..249
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             384..1178
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(438..457)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1048..1067
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1352..1473
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1389..1408)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1443..1462
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1522..1538)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1522..1538)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1535..1557)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1546..1562
                     /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(1570..1576,1577..1594,1595..1600))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1615..1636
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1753..1770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1924..2512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2004..2023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2683..3474,3475..3543))
                     /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     3306..3325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3544..3648)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3723..3742)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3914..4293
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4294..4497
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4448..4468
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4542..4561
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4542..4560
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(8366..8384)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(8367..8384)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(8404..8421)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8410..8634
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8680..9108
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8767..8786)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8977..8998
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(join(9117..9134,1..3))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(9122..9134,1..317)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
       61 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      121 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
      181 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
      241 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
      301 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag
      361 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
      421 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
      481 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
      541 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
      601 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
      661 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
      721 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
      781 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
      841 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
      901 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
      961 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     1021 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     1081 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     1141 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     1201 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     1261 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     1321 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     1381 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1441 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     1501 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1561 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1621 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1681 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1741 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1801 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1861 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1921 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1981 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2041 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2101 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2161 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     2221 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     2281 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     2341 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2401 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2461 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2521 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2581 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2641 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2701 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2761 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2821 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2881 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2941 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3001 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3061 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3121 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3181 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     3241 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     3301 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3361 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3421 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3481 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3541 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3601 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3661 aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc
     3721 agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg
     3781 gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac
     3841 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
     3901 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
     3961 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
     4021 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
     4081 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
     4141 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
     4201 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
     4261 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
     4321 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
     4381 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
     4441 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
     4501 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
     4561 gagacccaag cttggtaccg agctcggatc cactagtaac ggccgccagt gtgctggaat
     4621 tctgcagatc gcgaagatgg ctgctggctg cctgctggcc ttgactctga cacttttcca
     4681 atcttggctg atcggcccct cgagcgagga gcccttccct tcgcccgtca ctatcaagtc
     4741 atgggtggat aaaatgcagg aagaccttgt cacactggca aagacagcaa gtggagtgac
     4801 tcagcttgct gacatttatg aaaaatacca agatttgtat actgtggagc caaataatgc
     4861 acgccaactg gttgaaattg cagctagaga cattgagaaa cttctgagca acagatctaa
     4921 agccctggtg cgcctggcta tggaagcaga gaaagtccaa gccgcccacc aatggaggga
     4981 agattttgca agcaacgaag ttgtctacta caacgctaaa gacgatcttg atcctgaaag
     5041 aaatgagagt gagtcaggca gccaacggat taaacctgtt ttcatcgaag acgctaattt
     5101 tggacgccag atatcctatc aacatgcagc ggtccatatt cccacggaca tctatgaggg
     5161 ctcaaccata gtgttaaacg aactcaactg gacaagtgcc ttagatgaag ttttcaaaag
     5221 aaatcgagat gaagacccta cattgctgtg gcaagtgttt ggcagcgcca ccggcctggc
     5281 ccgatattat ccagcttctc cgtgggtgga taacagcaga actccaaaca agatcgatct
     5341 ctacgatgta cgcagaagac catggtacat ccagggagct gcatccccca aagacatgct
     5401 cattctggtg gacgtgagtg gaagtgtgag cggattgact ctgaaactca tccggacatc
     5461 tgtctctgag atgttagaaa ccctttccga tgatgatttc gtgaatgtag cttcatttaa
     5521 cagcaatgct caggatgtga gttgtttcca gcacctggtt caagcgaatg taaggaacaa
     5581 gaaggtgctg aaggatgcgg tgaataacat cacagcaaag gggataacag attacaagaa
     5641 aggctttagc ttcgcgtttg aacagctact taattataat gtttccagag ccaattgtaa
     5701 taagatcatc atgttattca cggatggagg agaagagaga gcccaggaga tatttgccaa
     5761 gtacaataaa gacaaaaaag tccgcgtgtt tacattttct gttggtcaac ataattatga
     5821 cagaggacct attcagtgga tggcttgtga aaacaaaggt tattattatg agattccctc
     5881 tattggtgca ataagaatca atactcagga atatctagat gttctgggaa gaccaatggt
     5941 tttagccggg gacaaagcaa agcaagtcca gtggacaaat gtgtatttgg atgctctgga
     6001 gctgggactt gtcattactg gaactctacc ggtcttcaac gtcactggcc agtctgaaaa
     6061 taagactaac ttgaagaacc agttgattct tggtgtgatg ggcgttgatg tgtctctgga
     6121 agatattaag agattgacac cacgttttac actgtgcccc aacggttact attttgcaat
     6181 cgatcctaat ggttatgtct tactgcatcc aaatcttcag ccaaagaacc ccaaatctca
     6241 ggagccggtc acactggatt ttctcgatgc tgagttagag aatgatatta aagtggagat
     6301 tcgaaataaa atgatagacg gagaaagtgg agaaaaaaca ttcagaactc tggtcaaatc
     6361 tcaagatgag agatatattg acaaaggaaa tcgaacatac acatggacac ctgtcaatgg
     6421 cacagattac agtttggcct tggtattgcc aacctacagt ttttactata taaaagccaa
     6481 aatagaagag acaataactc aggccagatc aaaaaagggg aaaatgaagg attcagaaac
     6541 cctgaagcca gacaattttg aagaatctgg ctacactttc atagcaccaa gggaatactg
     6601 caatgacctg aaaccttcag ataataacac tgaatttctt ttaaatttca atgaatttat
     6661 tgataggaaa actccaaaca acccttcctg taatacagat ttgattaata gaatcttgct
     6721 ggatgcgggt tttacaaatg aacttgtcca aaattattgg agtaaacaga aaaatatcaa
     6781 aggagtgaag gcacgctttg tggtgactga tggcggaatt acaagagttt accccaaaga
     6841 ggctggagaa aactggcaag agaacccaga gacgtatgag gacagcttct acaaacggag
     6901 cctagataac gataactacg ttttcactgc gccctacttc aacaaaagtg gacctggtgc
     6961 ctatgaatct ggaattatgg taagcaaagc tgtagaactg tacatccaag gaaaacttct
     7021 taagcctgca gttgtgggaa ttaaaattga cgtaaactcc tggatagaaa attttaccaa
     7081 aacttcaatc agggatccgt gtgctggtcc agtttgtgac tgcaaaagaa acagtgatgt
     7141 aatggactgt gtcattctag atgatggtgg atttcttctg atggccaatc atgatgatta
     7201 cactaatcag attggacgtt tttttggaga gattgacccg agcatgatga gacacctggt
     7261 taatatatca ctttatgcat tcaataaatc atatgactat cagtctgtgt gtgatccagg
     7321 ggcagcacca aagcaaggag caggacatcg ctcagcctat gtgccatcaa ttacagacat
     7381 actccagatt ggctggtggg ccactgctgc cgcctggtct attctccagc agctactctt
     7441 gagtttgaca tttccacggc tccttgaggc agttgaaatg gaagaggatg acttcacagc
     7501 ttccctgtct aagcagagct gcatcacaga acaaactcag tacttcttca agaacgatac
     7561 taaatcattc agtggtttac tggactgtgg aaactgttcc aggatctttc atgttgagaa
     7621 gcttatgaac accaacctag tgttcataat ggtggagagc aagggaacat gtccgtgtga
     7681 cacgcggctg ctcatgcaag cagaacagac ttctgatggt ccagatcctt gcgacatggt
     7741 taagcagccc agataccgaa aaggacctga tgtctgcttt gataacaatg tgctggagga
     7801 ttataccgac tgtggtggtg tttctggatt aaacccttcc ttatggtcta tctttggact
     7861 ccaatttata ctcctttggc tggtatctgg cagcagacac tatctatggt gaccctctaa
     7921 acccatgatc tgcaacttaa ttccatgccc tgccaaaaca tgagccctca gttacattaa
     7981 aatagagccc gttgaagaag ctccacagaa ccttagctgg gcctctaccc tgatgtaact
     8041 gtgaggctta gattcctgac catcgctggc tgcatgtcag ggtgttggaa actgaaactc
     8101 gtgtgaacat gctgcatcat ctgtgtccaa catcaaagca aagctctact cattctctgt
     8161 gggaggtttg ggagttcgct gttgcactgt tggagattac gtggaaaagg ctcccccacg
     8221 caggccttat gaaaagaagc agtcttgact ttgacctagc atttcaacct gctgcagttt
     8281 taccaagccg aattcctgca gcccggggga tccactagat ccatcacact ggcggccgct
     8341 cgagcatgca tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat
     8401 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     8461 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     8521 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     8581 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     8641 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     8701 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     8761 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     8821 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     8881 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     8941 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     9001 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     9061 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     9121 tgtgtgtcag ttag
//
