LOCUS       sequence-355948-        7806 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tDimer-tagged mitochondrial calcium uniporter (MCU) under 
            control of the CaMKIIa promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7806)
  AUTHORS   Qiu J, Tan YW, Hagenston AM, Martel MA, Kneisel N, Skehel PA, Wyllie
            DJ, Bading H, Hardingham GE
  TITLE     Mitochondrial calcium uniporter Mcu controls excitotoxicity and is 
            transcriptionally repressed by neuroprotective nuclear calcium 
            signals.
  JOURNAL   Nat Commun. 2013;4:2034. doi: 10.1038/ncomms3034.
  PUBMED    23774321
REFERENCE   2  (bases 1 to 7806)
  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: "Nat 
            Commun."; date: "2013"; volume: "4:2034. doi"; pages: " 
            10.1038/ncomms3034"
COMMENT     Sequence Label: pAAV-MCU-tDimer
FEATURES             Location/Qualifiers
     source          1..7806
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        147..1435
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1446..1455
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2505..2514
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2507..3184
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFL"
     CDS             3227..3901
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQA
                     LKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHL
                     FL"
     misc_feature    3935..4523
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3988..4008)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4406..4417)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4526..4542)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4527..4543)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4553..5029
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4752..4771)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5069..5209
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5069..5198
                     /label=AAV2 ITR
     rep_origin      5284..5739
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5371..5390)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5581..5602
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5756..5775)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5875..5897
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5935..5953)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6021..6125
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6126..6194,6195..6986)
                     /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     complement(6344..6363)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7157..7745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7646..7665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaacatt atggccttag gtcacttcat ctccatgggg
      181 ttcttcttct gattttctag aaaatgagat gggggtgcag agagcttcct cagtgacctg
      241 cccagggtca catcagaaat gtcagagcta gaacttgaac tcagattact aatcttaaat
      301 tccatgcctt gggggcatgc aagtacgata tacagaagga gtgaactcat tagggcagat
      361 gaccaatgag tttaggaaag aagagtccag ggcagggtac atctacacca cccgcccagc
      421 cctgggtgag tccagccacg ttcacctcat tatagttgcc tctctccagt cctaccttga
      481 cgggaagcac aagcagaaac tgggacagga gccccaggag accaaatctt catggtccct
      541 ctgggaggat gggtggggag agctgtggca gaggcctcag gaggggccct gctgctcagt
      601 ggtgacagat aggggtgaga aagcagacag agtcattccg tcagcattct gggtctgttt
      661 ggtacttctt ctcacgctaa ggtggcggtg tgatatgcac aatggctaaa aagcagggag
      721 agctggaaag aaacaaggac agagacagag gccaagtcaa ccagaccaat tcccagagga
      781 agcaaagaaa ccattacaga gactacaagg gggaagggaa ggagagatga attagcttcc
      841 cctgtaaacc ttagaaccca gctgttgcca gggcaacggg gcaatacctg tctcttcaga
      901 ggagatgaag ttgccagggt aactacatcc tgtctttctc aaggaccatc ccagaatgtg
      961 gcacccacta gccgttacca tagcaactgc ctctttgccc cacttaatcc catcccgtct
     1021 gttaaaaggg ccctatagtt ggaggtgggg gaggtaggaa gagcgatgat cacttgtgga
     1081 ctaagtttgt tcgcatcccc ttctccaacc ccctcagtac atcaccctgg gggaacaggg
     1141 tccacttgct cctgggccca cacagtcctg cagtattgtg tatataaggc cagggcaaag
     1201 aggagcaggt tttaaagtga aaggcaggca ggtgttgggg aggcagttac cggggcaacg
     1261 ggaacagggc gtttcggagg tggttgccat ggggacctgg atgctgacga aggctcgcga
     1321 ggctgtgagc agccacagtg ccctgctcag aagccccaag ctcgtcagtc aagccggttc
     1381 tccgtttgca ctcaggagca cgggcaggcg agtggcccct agttctgggg gcagcggggg
     1441 atccaccatg gcggccgccg caggtagatc gctcctgctg ctgctctgct cccggggcgg
     1501 cggcgggggc gccggcggct gcggagcgct gactgccggc tgctttcccg ggctgggcgt
     1561 cagccgccac cggccgcacc agcagcaccg gacggcacac cagaggccgg cttcctggca
     1621 gagcgtggga gccgcatatt gcagtacggt tgtgccctct gatgacgtga cggtggttta
     1681 ccaaaatgga ttaccggtga tatctgtgag gctaccctct cgacgcgagc gctgccaatt
     1741 cacactcaag cctatctctg actcagtcgg cgtgttttta cgacaactgc aagaggagga
     1801 tcggggaatc gaccgggtcg ccatctactc accagatggc gttcgagtcg ctgcctccac
     1861 ggggatagac ctcctgctcc tcgatgactt taagcttgtc attaatgact taacatacca
     1921 cgtacggcca ccaaagagag acctcctaag ccatgaagat gcagcgacgc tgaacgacgt
     1981 gaagaccctg gtccagcagc tgtacaccac actgtgcatt gagcagcatc agcttaacaa
     2041 agagcgggag ctcgtggaga ggttagagga cctcaagcag cagctggccc ccctggagaa
     2101 ggtacgaatt gaaattagca gaaaagcaga gaagaggacc actctggtgc tgtggggagg
     2161 cctggcctac atggccaccc agtttggcat tctggcccgg ctcacctggt gggagtactc
     2221 gtgggacatc atggagcccg tcacctactt catcacgtac ggaagcgcca tggccatgta
     2281 tgcgtatttt gtaatgacgc gccaggaata tgtttatcca gaagccagag acagacaata
     2341 cttattattt ttccataaag gagccaaaaa gtcacgtttc gacctagaga aatacaatca
     2401 gctcaaggat gcaattgctc aggcagaaat ggatcttaag agactgagag acccattaca
     2461 agtacacctg cccctccgac agatcggaga aaaggaagct agcaccatgg tggcctcctc
     2521 cgaggacgtc atcaaagagt tcatgcgctt caaggtgcgc atggagggct ccgtgaacgg
     2581 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
     2641 caagctgaag gtgaccaagg gcggccccct gcccttcgcc tgggacatcc tgtcccccca
     2701 gttccagtac ggctccaagg cgtacgtgaa gcaccccgcc gacatccccg actacaagaa
     2761 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
     2821 ggtgaccgtg acccaggact cctccctgca ggacggcacg ctgatctaca aggtgaagtt
     2881 ccgcggcacc aacttccccc ccgacggccc cgtaatgcag aagaagacca tgggctggga
     2941 ggcctccacc gagcgcctgt acccccgcga cggcgtgctg aagggcgaga tccaccaggc
     3001 cctgaagctg aaggacggcg gccactacct ggtggagttc aagaccatct acatggccaa
     3061 gaagcccgtg cagctgcccg gctactacta cgtggacacc aagctggaca tcacctccca
     3121 caacgaggac tacaccatcg tggaacagta cgagcgctcc gagggccgcc accacctgtt
     3181 cctggggcat ggcaccggca gcaccggcag cggcagctcc ggcaccgcct cctccgagga
     3241 cgtcatcaaa gagttcatgc gcttcaaggt gcgcatggag ggctccgtga acggccacga
     3301 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     3361 gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc cccagttcca
     3421 gtacggctcc aaggcgtacg tgaagcaccc cgccgacatc cccgactaca agaagctgtc
     3481 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     3541 cgtgacccag gactcctccc tgcaggacgg cacgctgatc tacaaggtga agttccgcgg
     3601 caccaacttc ccccccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     3661 caccgagcgc ctgtaccccc gcgacggcgt gctgaagggc gagatccacc aggccctgaa
     3721 gctgaaggac ggcggccact acctggtgga gttcaagacc atctacatgg ccaagaagcc
     3781 cgtgcagctg cccggctact actacgtgga caccaagctg gacatcacct cccacaacga
     3841 ggactacacc atcgtggaac agtacgagcg ctccgagggc cgccaccacc tgttcctgta
     3901 ggtcgacttc gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg
     3961 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     4021 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     4081 taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt
     4141 ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca
     4201 gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc
     4261 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4321 gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg
     4381 cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg
     4441 cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat
     4501 ctccctttgg gccgcctccc cgcatcgata ccgtcgacct cgagagatct acgggtggca
     4561 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     4621 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     4681 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     4741 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     4801 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     4861 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     4921 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     4981 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     5041 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     5101 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     5161 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     5221 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     5281 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5341 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5401 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5461 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     5521 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5581 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     5641 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     5701 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     5761 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5821 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5881 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5941 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     6001 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6061 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6121 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6181 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6241 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6301 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6361 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6421 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6481 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6541 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6601 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6661 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6721 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6781 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6841 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6901 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6961 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     7021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7741 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7801 acatgt
//
