LOCUS       sequence-415382-        9129 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 and with R421A and D491A 
            mutations that both disrupt Gabapentin binding.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9129)
  TITLE     Dolphin Lab Plasmids 2023
REFERENCE   2  (bases 1 to 9129)
  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 R241A D491A HA pcDNA3
FEATURES             Location/Qualifiers
     source          1..9129
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      127..262
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     189..208
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             343..1137
                     /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(397..416)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1007..1026
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1311..1432
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1348..1367)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1402..1421
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1481..1497)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1481..1497)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1494..1516)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1505..1521
                     /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(1529..1535,1536..1553,1554..1559))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1574..1595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1712..1729)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1883..2471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1963..1982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2642..3433,3434..3502))
                     /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     3265..3284
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3503..3607)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3682..3701)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3873..4252
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4253..4456
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4407..4427
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4501..4520
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4501..4519
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             6243..6269
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(8320..8338)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(8321..8338)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(8358..8375)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8364..8588
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8634..9062
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8721..8740)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8931..8952
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(9071..9091)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(9076..9129,1..276)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
       61 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
      121 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
      181 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
      241 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
      301 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
      361 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
      421 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
      481 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
      541 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
      601 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
      661 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
      721 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
      781 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
      841 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
      901 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
      961 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     1021 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     1081 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     1141 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     1201 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     1261 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     1321 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     1381 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     1441 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1501 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1561 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1621 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1681 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1741 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1801 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1861 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1921 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1981 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2041 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2101 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2161 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2221 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2281 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2341 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2401 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2461 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2521 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2581 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2641 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2701 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2761 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2821 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2881 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2941 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3001 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3061 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3121 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3181 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3241 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3301 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3361 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3421 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3481 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3541 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3601 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     3661 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     3721 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     3781 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     3841 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
     3901 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     3961 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     4021 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     4081 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     4141 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     4201 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     4261 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     4321 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     4381 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     4441 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
     4501 taatacgact cactataggg agacccaagc ttggtaccga gctcggatcc actagtaacg
     4561 gccgccagtg tgctggaatt ctgcagatcg cgaagatggc tgctggctgc ctgctggcct
     4621 tgactctgac acttttccaa tcttggctga tcggcccctc gagcgaggag cccttccctt
     4681 cgcccgtcac tatcaagtca tgggtggata aaatgcagga agaccttgtc acactggcaa
     4741 agacagcaag tggagtgact cagcttgctg acatttatga aaaataccaa gatttgtata
     4801 ctgtggagcc aaataatgca cgccaactgg ttgaaattgc agctagagac attgagaaac
     4861 ttctgagcaa cagatctaaa gccctggtgc gcctggctat ggaagcagag aaagtccaag
     4921 ccgcccacca atggagggaa gattttgcaa gcaacgaagt tgtctactac aacgctaaag
     4981 acgatcttga tcctgaaaga aatgagagtg agtcaggcag ccaacggatt aaacctgttt
     5041 tcatcgaaga cgctaatttt ggacgccaga tatcctatca acatgcagcg gtccatattc
     5101 ccacggacat ctatgagggc tcaaccatag tgttaaacga actcaactgg acaagtgcct
     5161 tagatgaagt tttcaaaaga aatcgagatg aagaccctac attgctgtgg caagtgtttg
     5221 gcagcgccac cggcctggcc cgatattatc cagcttctcc gtgggtggat aacagcagaa
     5281 ctccaaacaa gatcgatctc tacgatgtac gcagagcacc atggtacatc cagggagctg
     5341 catcccccaa agacatgctc attctggtgg acgtgagtgg aagtgtgagc ggattgactc
     5401 tgaaactcat ccggacatct gtctctgaga tgttagaaac cctttccgat gatgatttcg
     5461 tgaatgtagc ttcatttaac agcaatgctc aggatgtgag ttgtttccag cacctggttc
     5521 aagcgaatgt aaggaacaag aaggtgctga aggatgcggt gaataacatc acagcaaagg
     5581 ggataacaga ttacaagaaa ggctttagct tcgcgtttga acagctactt aattataatg
     5641 tttccagagc caattgtaat aagatcatca tgttattcac ggatggagga gaagagagag
     5701 cccaggagat atttgccaag tacaataaag acaaaaaagt ccgcgtgttt acattttctg
     5761 ttggtcaaca taattatgac agaggaccta ttcagtggat ggcttgtgaa aacaaaggtt
     5821 attattatga gattccctct attggtgcaa taagaatcaa tactcaggaa tatctagatg
     5881 ttctgggaag accaatggtt ttagccgggg acaaagcaaa gcaagtccag tggacaaatg
     5941 tgtatttgga tgctctggag ctgggacttg tcattactgg aactctaccg gtcttcaacg
     6001 tcactggcca gtctgaaaat aagactaact tgaagaacca gttgattctt ggtgtgatgg
     6061 gcgttgctgt gtctctggaa gatattaaga gattgacacc acgttttaca ctgtgcccca
     6121 acggttacta ttttgcaatc gatcctaatg gttatgtctt actgcatcca aatcttcagc
     6181 caaagaaccc caaatctcag gagccggtca cactggattt tctcgatgct gagttagaga
     6241 attaccctta cgacgtccct gactacgccg atattaaagt ggagattcga aataaaatga
     6301 tagacggaga aagtggagaa aaaacattca gaactctggt caaatctcaa gatgagagat
     6361 atattgacaa aggaaatcga acatacacat ggacacctgt caatggcaca gattacagtt
     6421 tggccttggt attgccaacc tacagttttt actatataaa agccaaaata gaagagacaa
     6481 taactcaggc cagatcaaaa aaggggaaaa tgaaggattc agaaaccctg aagccagaca
     6541 attttgaaga atctggctac actttcatag caccaaggga atactgcaat gacctgaaac
     6601 cttcagataa taacactgaa tttcttttaa atttcaatga atttattgat aggaaaactc
     6661 caaacaaccc ttcctgtaat acagatttga ttaatagaat cttgctggat gcgggtttta
     6721 caaatgaact tgtccaaaat tattggagta aacagaaaaa tatcaaagga gtgaaggcac
     6781 gctttgtggt gactgatggc ggaattacaa gagtttaccc caaagaggct ggagaaaact
     6841 ggcaagagaa cccagagacg tatgaggaca gcttctacaa acggagccta gataacgata
     6901 actacgtttt cactgcgccc tacttcaaca aaagtggacc tggtgcctat gaatctggaa
     6961 ttatggtaag caaagctgta gaactgtaca tccaaggaaa acttcttaag cctgcagttg
     7021 tgggaattaa aattgacgta aactcctgga tagaaaattt taccaaaact tcaatcaggg
     7081 atccgtgtgc tggtccagtt tgtgactgca aaagaaacag tgatgtaatg gactgtgtca
     7141 ttctagatga tggtggattt cttctgatgg ccaatcatga tgattacact aatcagattg
     7201 gacgtttttt tggagagatt gacccgagca tgatgagaca cctggttaat atatcacttt
     7261 atgcattcaa taaatcatat gactatcagt ctgtgtgtga tccaggggca gcaccaaagc
     7321 aaggagcagg acatcgctca gcctatgtgc catcaattac agacatactc cagattggct
     7381 ggtgggccac tgctgccgcc tggtctattc tccagcagct actcttgagt ttgacatttc
     7441 cacggctcct tgaggcagtt gaaatggaag aggatgactt cacagcttcc ctgtctaagc
     7501 agagctgcat cacagaacaa actcagtact tcttcaagaa cgatactaaa tcattcagtg
     7561 gtttactgga ctgtggaaac tgttccagga tctttcatgt tgagaagctt atgaacacca
     7621 acctagtgtt cataatggtg gagagcaagg gaacatgtcc gtgtgacacg cggctgctca
     7681 tgcaagcaga acagacttct gatggtccag atccttgcga catggttaag cagcccagat
     7741 accgaaaagg acctgatgtc tgctttgata acaatgtgct ggaggattat accgactgtg
     7801 gtggtgtttc tggattaaac ccttccttat ggtctatctt tggactccaa tttatactcc
     7861 tttggctggt atctggcagc agacactatc tatggtgacc ctctaaaccc atgatctgca
     7921 acttaattcc atgccctgcc aaaacatgag ccctcagtta cattaaaata gagcccgttg
     7981 aagaagctcc acagaacctt agctgggcct ctaccctgat gtaactgtga ggcttagatt
     8041 cctgaccatc gctggctgca tgtcagggtg ttggaaactg aaactcgtgt gaacatgctg
     8101 catcatctgt gtccaacatc aaagcaaagc tctactcatt ctctgtggga ggtttgggag
     8161 ttcgctgttg cactgttgga gattacgtgg aaaaggctcc cccacgcagg ccttatgaaa
     8221 agaagcagtc ttgactttga cctagcattt caacctgctg cagttttacc aagccgaatt
     8281 cgcatgcggc cgctcgagca tgcatctaga gggccctatt ctatagtgtc acctaaatgc
     8341 tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     8401 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     8461 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     8521 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     8581 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     8641 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     8701 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     8761 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     8821 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     8881 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     8941 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     9001 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     9061 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     9121 agaagtatg
//
