LOCUS       sequence-427350-        5875 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transfer plasmid for AAV vector production of mCherry and miR for 
            rat Rn ATP10B.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5875)
  AUTHORS   Sanchiz-Calvo M, Coccia E, Cawthorne C, Morrone Parfitt G, 
            Torre-Muruzabal T, Tsafaras G, Van Laere K, Cabezudo D, Cascalho A, 
            Van den Haute C, Vangheluwe P, Blanchard J, Bentea E, Baekelandt V
  TITLE     Loss of the lysosomal lipid flippase ATP10B leads to progressive 
            dopaminergic neurodegeneration and parkinsonian motor deficits.
  JOURNAL   Acta Neuropathol. 2025 Jul 17;150(1):5. doi: 
            10.1007/s00401-025-02908-0.
  PUBMED    40676227
REFERENCE   2  (bases 1 to 5875)
  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: "Acta 
            Neuropathol."; date: "2025-07-17"; volume: "150(1):5. doi"; pages: "
            10.1007/s00401-025-02908-0"
COMMENT     Sequence Label: pAAV-CMVenh synapsin-intron-mCherry-1x miR7 Rn...
FEATURES             Location/Qualifiers
     source          1..5875
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        282..585
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1020..1152
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1135..1154
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1189..1198
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1195..1905
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1340..1358)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1608..1627
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     ncRNA           1987..2001
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     primer_bind     complement(2139..2159)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2557..2568)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2635..2652)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2641..2848
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2936..3065
                     /label=AAV2 ITR
     primer_bind     complement(3084..3101)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3084..3100)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3093..3115)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3242..3697
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3329..3348)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3539..3560
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3723..3827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3828..3896,3897..4688)
                     /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(4046..4065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4859..5447
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5348..5367
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5735..5756
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5771..5776,5777..5794,5795..5801)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5809..5825
                     /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)."
     primer_bind     5814..5836
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5833..5849
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5833..5849
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
      181 agccatgctc taggaagatc ggaattcgcc cttaagctag atagttatta atagtaatca
      241 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      301 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      361 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      421 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      481 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      541 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggcacc tgcagagggc
      601 cctgcgtatg agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc
      661 tgacgaccga ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc
      721 ccctatcaga gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt
      781 cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca
      841 ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg
      901 gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg
      961 ggcacgggcg cgaccatctg cgctgcggcc gcactagttg gtcgtgaggc actgggcagg
     1021 taagtatcaa ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac
     1081 agagaagact cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct
     1141 ttctctccac aggtgtccag gcggccgctc tagaggatcc ccaccggtgc caccatggtg
     1201 agcaagggcg aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1261 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1321 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
     1381 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     1441 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1501 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1561 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     1621 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     1681 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
     1741 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     1801 aagttggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc
     1861 gagggccgcc actccaccgg cggcatggac gagctgtaca agtaaagcgg ccgcgactct
     1921 agctcgagat ccaagaaggt atattgctgt tgacagtgag cgacctaaga cagtgcctat
     1981 atattagtga agccacagat gtaatatata ggcactgtct taggatgcct actgcctcgg
     2041 acttcaaggg ctacgatgtc gacttggatc catcgcgcgt gatccaatca acctctggat
     2101 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     2161 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     2221 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     2281 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     2341 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     2401 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     2461 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     2521 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     2581 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgaga tctgcctcga
     2641 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     2701 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     2761 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     2821 gggaagacaa tagcaggcat gctggggact cgagttaagg gcgaattccc gataaggatc
     2881 ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa
     2941 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3001 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     3061 cgcagcctta attaacctaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     3121 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     3181 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     3241 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3301 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3361 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3421 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3481 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3541 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3601 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3661 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     3721 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3781 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3841 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3901 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3961 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4021 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4081 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4141 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4201 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4261 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4321 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4381 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4441 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4501 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4561 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4621 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4681 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     4741 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4801 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4861 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4921 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4981 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5041 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5101 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5161 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5221 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5281 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5341 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5401 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5461 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5521 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5581 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     5641 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     5701 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     5761 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     5821 aacaatttca cacaggaaac agctatgacc atgattacgc cagatttaat taagg
//
