LOCUS       sequence-415391-        8750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of rat alpha2delta-1 calcium channel auxillary subunit 
            with a single Bungarotoxin Binding Site motif inserted after 
            aa549HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8750)
  AUTHORS   Meyer JO, Dolphin AC
  TITLE     Rab11-dependent recycling of calcium channels is mediated by 
            auxiliary subunit alpha(2)delta-1 but not alpha(2)delta-3.
  JOURNAL   Sci Rep. 2021 May 13;11(1):10256. doi: 10.1038/s41598-021-89820-1.
  PUBMED    33986433
REFERENCE   2  (bases 1 to 8750)
  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: "Sci Rep."; 
            date: "2021-05-13"; volume: "11(1):10256. doi"; pages: " 
            10.1038/s41598-021-89820-1"
COMMENT     Sequence Label: cacna2d1 BBS pcDNA3
FEATURES             Location/Qualifiers
     source          1..8750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      175..310
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     237..256
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        372..419
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             438..812
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    942..1063
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(979..998)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1033..1052
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1112..1128)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1112..1128)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1125..1147)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1136..1152
                     /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(1160..1166,1167..1184,1185..1190))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1205..1226
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1343..1360)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1514..2102)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1594..1613)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2273..3064,3065..3133))
                     /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     2896..2915
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3134..3238)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3313..3332)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3504..3883
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3884..4087
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4038..4058
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4132..4151
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4132..4150
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8027..8044)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    8033..8257
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      8303..8731
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8390..8409)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8600..8621
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(join(8740..8750,1..10))
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(8745..8750,1..324)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
       61 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      121 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
      181 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
      241 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
      301 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
      361 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
      421 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
      481 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
      541 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
      601 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
      661 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
      721 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
      781 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
      841 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
      901 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
      961 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     1021 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     1081 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     1141 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     1201 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     1261 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1321 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1381 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1441 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1501 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1561 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1621 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1681 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1741 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1801 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1861 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1921 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1981 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2041 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2101 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2161 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2221 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2281 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2341 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2401 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2461 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2521 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2581 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2641 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2701 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2761 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2821 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2881 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2941 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3001 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3061 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3121 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3181 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3241 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     3301 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     3361 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     3421 cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga
     3481 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
     3541 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     3601 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     3661 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     3721 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
     3781 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
     3841 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     3901 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     3961 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     4021 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     4081 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
     4141 tcactatagg gagacccaag ctggctagcg tttaaactta agcttggtac cgagctcgga
     4201 tccactagta acggccgcca gtgtgctgga attctgcaga tcgcgaagat ggctgctggc
     4261 tgcctgctgg ccttgactct gacacttttc caatcttggc tgatcggccc ctcgagcgag
     4321 gagcccttcc cttcgcccgt cactatcaag tcatgggtgg ataaaatgca ggaagacctt
     4381 gtcacactgg caaagacagc aagtggagtg actcagcttg ctgacattta tgaaaaatac
     4441 caagatttgt atactgtgga gccaaataat gcacgccaac tggttgaaat tgcagctaga
     4501 gacattgaga aacttctgag caacagatct aaagccctgg tgcgcctggc tatggaagca
     4561 gagaaagtcc aagccgccca ccaatggagg gaagattttg caagcaacga agttgtctac
     4621 tacaacgcta aagacgatct tgatcctgaa agaaatgaga gtgagtcagg cagccaacgg
     4681 attaaacctg ttttcatcga agacgctaat tttggacgcc agatatccta tcaacatgca
     4741 gcggtccata ttcccacgga catctatgag ggctcaacca tagtgttaaa cgaactcaac
     4801 tggacaagtg ccttagatga agttttcaaa agaaatcgag atgaagaccc tacattgctg
     4861 tggcaagtgt ttggcagcgc caccggcctg gcccgatatt atccagcttc tccgtgggtg
     4921 gataacagca gaactccaaa caagatcgat ctctacgatg tacgcagaag accatggtac
     4981 atccagggag ctgcatcccc caaagacatg ctcattctgg tggacgtgag tggaagtgtg
     5041 agcggattga ctctgaaact catccggaca tctgtctctg agatgttaga aaccctttcc
     5101 gatgatgatt tcgtgaatgt agcttcattt aacagcaatg ctcaggatgt gagttgtttc
     5161 cagcacctgg ttcaagcgaa tgtaaggaac aagaaggtgc tgaaggatgc ggtgaataac
     5221 atcacagcaa aggggataac agattacaag aaaggcttta gcttcgcgtt tgaacagcta
     5281 cttaattata atgtttccag agccaattgt aataagatca tcatgttatt cacggatgga
     5341 ggagaagaga gagcccagga gatatttgcc aagtacaata aagacaaaaa agtccgcgtg
     5401 tttacatttt ctgttggtca acataattat gacagaggac ctattcagtg gatggcttgt
     5461 gaaaacaaag gttattatta tgagattccc tctattggtg caataagaat caatactcag
     5521 gaatatctag atgttctggg aagaccaatg gttttagccg gggacaaagc aaagcaagtc
     5581 cagtggacaa atgtgtattt ggatgctctg gagctgggac ttgtcattac tggaactcta
     5641 ccggtcttca acgtcactgg ccagtctgaa aataagacta acttgaagaa ccagttgatt
     5701 cttggtgtga tgggcgttga tgtgtctctg gaagatatta agagattgac accacgtttt
     5761 acactgtgcc ccaacggtta ctattttgca atcgatccta atggttatgt cttactgcat
     5821 ccaaatcttc agccaaagaa ccccaaatct caggagccgg tcacactgga ttttctcgat
     5881 gctgagttag agaattggag atactacgag agctccctgg agccctaccc tgacgatatt
     5941 aaagtggaga ttcgaaataa aatgatagac ggagaaagtg gagaaaaaac attcagaact
     6001 ctggtcaaat ctcaagatga gagatatatt gacaaaggaa atcgaacata cacatggaca
     6061 cctgtcaatg gcacagatta cagtttggcc ttggtattgc caacctacag tttttactat
     6121 ataaaagcca aaatagaaga gacaataact caggccagat caaaaaaggg gaaaatgaag
     6181 gattcagaaa ccctgaagcc agacaatttt gaagaatctg gctacacttt catagcacca
     6241 agggaatact gcaatgacct gaaaccttca gataataaca ctgaatttct tttaaatttc
     6301 aatgaattta ttgataggaa aactccaaac aacccttcct gtaatacaga tttgattaat
     6361 agaatcttgc tggatgcggg ttttacaaat gaacttgtcc aaaattattg gagtaaacag
     6421 aaaaatatca aaggagtgaa ggcacgcttt gtggtgactg atggcggaat tacaagagtt
     6481 taccccaaag aggctggaga aaactggcaa gagaacccag agacgtatga ggacagcttc
     6541 tacaaacgga gcctagataa cgataactac gttttcactg cgccctactt caacaaaagt
     6601 ggacctggtg cctatgaatc tggaattatg gtaagcaaag ctgtagaact gtacatccaa
     6661 ggaaaacttc ttaagcctgc agttgtggga attaaaattg acgtaaactc ctggatagaa
     6721 aattttacca aaacttcaat cagggatccg tgtgctggtc cagtttgtga ctgcaaaaga
     6781 aacagtgatg taatggactg tgtcattcta gatgatggtg gatttcttct gatggccaat
     6841 catgatgatt acactaatca gattggacgt ttttttggag agattgaccc gagcatgatg
     6901 agacacctgg ttaatatatc actttatgca ttcaataaat catatgacta tcagtctgtg
     6961 tgtgatccag gggcagcacc aaagcaagga gcaggacatc gctcagccta tgtgccatca
     7021 attacagaca tactccagat tggctggtgg gccactgctg ccgcctggtc tattctccag
     7081 cagctactct tgagtttgac atttccacgg ctccttgagg cagttgaaat ggaagaggat
     7141 gacttcacag cttccctgtc taagcagagc tgcatcacag aacaaactca gtacttcttc
     7201 aagaacgata ctaaatcatt cagtggttta ctggactgtg gaaactgttc caggatcttt
     7261 catgttgaga agcttatgaa caccaaccta gtgttcataa tggtggagag caagggaaca
     7321 tgtccgtgtg acacgcggct gctcatgcaa gcagaacaga cttctgatgg tccagatcct
     7381 tgcgacatgg ttaagcagcc cagataccga aaaggacctg atgtctgctt tgataacaat
     7441 gtgctggagg attataccga ctgtggtggt gtttctggat taaacccttc cttatggtct
     7501 atctttggac tccaatttat actcctttgg ctggtatctg gcagcagaca ctatctatgg
     7561 tgaccctcta aacccatgat ctgcaactta attccatgcc ctgccaaaac atgagccctc
     7621 agttacatta aaatagagcc cgttgaagaa gctccacaga accttagctg ggcctctacc
     7681 ctgatgtaac tgtgaggctt agattcctga ccatcgctgg ctgcatgtca gggtgttgga
     7741 aactgaaact cgtgtgaaca tgctgcatca tctgtgtcca acatcaaagc aaagctctac
     7801 tcattctctg tgggaggttt gggagttcgc tgttgcactg ttggagatta cgtggaaaag
     7861 gctcccccac gcaggcctta tgaaaagaag cagtcttgac tttgacctag catttcaacc
     7921 tgctgcagtt ttaccaagcc gaattcctgc agcccggggg atccactaga tccatcacac
     7981 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     8041 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     8101 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     8161 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     8221 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     8281 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     8341 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     8401 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     8461 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     8521 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     8581 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     8641 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     8701 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
//
