LOCUS       sequence-355942-        7119 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses GFP-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 7119)
  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 7119)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-EGFP
FEATURES             Location/Qualifiers
     source          1..7119
                     /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"
     CDS             join(2504..2506,2507..2509,2510..3223)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2504..2506,2507..2509,2510..3220)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2549..2570)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2810..2829)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3157..3178
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3248..3836
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3301..3321)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3719..3730)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3839..3855)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3840..3856)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3866..4342
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4065..4084)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4382..4522
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4382..4511
                     /label=AAV2 ITR
     rep_origin      4597..5052
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4684..4703)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4894..4915
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5069..5088)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5188..5210
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5248..5266)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5334..5438
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5439..5507,5508..6299)
                     /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(5657..5676)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6470..7058
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6959..6978
                     /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 agcatggtga gcaagggcga
     2521 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2581 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2641 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2701 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2761 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     2821 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2881 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2941 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     3001 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3061 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3121 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3181 cgccgccggg atcactctcg gcatggacga gctgtacaag taagaattcg atatcaagct
     3241 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     3301 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     3361 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     3421 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3481 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3541 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3601 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3661 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3721 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3781 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3841 ataccgtcga cctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct
     3901 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     3961 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     4021 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     4081 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     4141 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     4201 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     4261 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     4321 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg
     4381 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4441 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     4501 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     4561 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     4621 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc
     4681 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4741 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4801 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4861 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4921 cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct
     4981 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     5041 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5101 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     5161 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     5221 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     5281 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5341 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5401 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5461 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5521 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5581 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5641 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5701 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5761 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5821 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5881 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5941 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     6001 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     6061 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6121 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6181 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6241 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6301 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6361 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6421 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6481 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6541 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6601 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6661 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6721 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6781 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6841 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6901 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6961 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     7021 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     7081 tacggttcct ggccttttgc tggccttttg ctcacatgt
//
