LOCUS       sequence-427351-        5858 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  transfer plasmid for AAV vector production of mCherry and miR 
            scramble (no target).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5858)
  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 5858)
  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 miR30 SCR
FEATURES             Location/Qualifiers
     source          1..5858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   7..136
                     /label=AAV2 ITR
     enhancer        277..580
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          1015..1147
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1130..1149
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1184..1193
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1190..1900
                     /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(1335..1353)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1603..1622
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     ncRNA           1965..1979
                     /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(2117..2137)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2535..2546)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2613..2630)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2619..2826
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2914..3043
                     /label=AAV2 ITR
     primer_bind     complement(3062..3079)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3062..3078)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3071..3093)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3220..3675
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3307..3326)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3517..3538
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3701..3805
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3806..3874,3875..4666)
                     /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(4024..4043)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4837..5425
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5326..5345
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5713..5734
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5749..5754,5755..5772,5773..5779)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5787..5803
                     /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     5792..5814
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5811..5827
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5811..5827
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 attaggctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg
       61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
      121 atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca
      181 tgctctagga agatcggaat tcgcccttaa gctagatagt tattaatagt aatcaattac
      241 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      301 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      361 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      421 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      481 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      541 ttggcagtac atctacgtat tagtcatcgc tattaccatg gcacctgcag agggccctgc
      601 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      661 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      721 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      781 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      841 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      901 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      961 gggcgcgacc atctgcgctg cggccgcact agttggtcgt gaggcactgg gcaggtaagt
     1021 atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc gagacagaga
     1081 agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt tgcctttctc
     1141 tccacaggtg tccaggcggc cgctctagag gatccccacc ggtgccacca tggtgagcaa
     1201 gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga
     1261 gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga
     1321 gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga
     1381 catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat
     1441 ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt
     1501 cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat
     1561 ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa
     1621 gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg
     1681 cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac
     1741 cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt
     1801 ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg
     1861 ccgccactcc accggcggca tggacgagct gtacaagtaa ctcgagatcc aagaaggtat
     1921 attgctgttg acagtgagcg aatacgacgg taagtgagta cgtagtgaag ccacagatgt
     1981 acgtactcac ttaccgtcgt atctgcctac tgcctcggac ttcaagggct acgatgtcga
     2041 cttggatcca tcgcgcgtga tccaatcaac ctctggatta caaaatttgt gaaagattga
     2101 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2161 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2221 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     2281 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     2341 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     2401 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     2461 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     2521 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     2581 ctctgcggcc tcttccgcgt cttcgagatc tgcctcgact gtgccttcta gttgccagcc
     2641 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2701 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2761 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     2821 tggggactcg agttaagggc gaattcccga taaggatctt cctagagcat ggctacgtag
     2881 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     2941 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     3001 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagccttaat taacctaatt
     3061 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     3121 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     3181 gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat
     3241 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     3301 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     3361 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     3421 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     3481 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     3541 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     3601 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     3661 taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     3721 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     3781 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     3841 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     3901 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     3961 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4021 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4081 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4141 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     4201 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     4261 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     4321 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     4381 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     4441 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     4501 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     4561 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     4621 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     4681 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     4741 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     4801 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     4861 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     4921 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     4981 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5041 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5101 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5161 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5221 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5281 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5341 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     5401 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     5461 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     5521 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     5581 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     5641 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
     5701 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     5761 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     5821 ctatgaccat gattacgcca gatttaatta aggcctta
//
