LOCUS       sequence-436347-        6250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Neuronal specific expression of the fusion construct PGK1-mRuby.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6250)
  AUTHORS   Kokotos AC, Antoniazzi AM, Unda SR, Ko MS, Park D, Eliezer D, 
            Kaplitt MG, De Camilli P, Ryan TA
  TITLE     Phosphoglycerate kinase is a central leverage point in Parkinson's 
            disease-driven neuronal metabolic deficits.
  JOURNAL   Sci Adv. 2024 Aug 23;10(34):eadn6016. doi: 10.1126/sciadv.adn6016. 
            Epub 2024 Aug 21.
  PUBMED    39167658
REFERENCE   2  (bases 1 to 6250)
  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 Adv."; 
            date: "2024-08-23"; volume: "10(34):eadn6016. doi"; pages: " 
            10.1126/sciadv.adn6016. Epub 2024 Aug 21"
COMMENT     Sequence Label: AAV hSyn hPGK1-mRuby
FEATURES             Location/Qualifiers
     source          1..6250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   95..224
                     /label=AAV2 ITR
     promoter        306..753
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             2056..2766
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTD
                     IALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAV
                     AKYSNLGGGMDELYK"
     misc_feature    2782..3370
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2835..2855)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3253..3264)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3373..3389)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3374..3390)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3419..3540
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3430..3449)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3484..3503
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3702..3831
                     /label=AAV2 ITR
     rep_origin      complement(4068..4656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4148..4167)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4827..5618,5619..5687))
                     /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     5450..5469
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5688..5792)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(5818..6250,1..23))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5955..5976)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6167..6186
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg
       61 gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc
      121 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      181 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa
      241 cccgccatgc tacttatcta cgtagccatg ctctaggaag atctctgcag agggccctgc
      301 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      361 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      421 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      481 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      541 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      601 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      661 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      721 agcggaggag tcgtgtcgtg cctgagagcg cagtcgaatt caagctgcta gcaaggatcc
      781 gccaccatgt cgctttctaa caagctgacg ctggacaagc tggacgttaa agggaagcgg
      841 gtcgttatga gagtcgactt caatgttcct atgaagaaca accagataac aaacaaccag
      901 aggattaagg ctgctgtccc aagcatcaaa ttctgcttgg acaatggagc caagtcggta
      961 gtccttatga gccacctagg ccggcctgat ggtgtgccca tgcctgacaa gtactcctta
     1021 gagccagttg ctgtagaact caaatctctg ctgggcaagg atgttctgtt cttgaaggac
     1081 tgtgtaggcc cagaagtgga gaaagcctgt gccaacccag ctgctgggtc tgtcatcctg
     1141 ctggagaacc tccgctttca tgtggaggaa gaagggaagg gaaaagatgc ttctgggaac
     1201 aaggttaaag ccgagccagc caaaatagaa gctttccgag cttcactttc caagctaggg
     1261 gatgtctatg tcaatgatgc ttttggcact gctcacagag cccacagctc catggtagga
     1321 gtcaatctgc cacagaaggc tggtgggttt ttgatgaaga aggagctgaa ctactttgca
     1381 aaggccttgg agagcccaga gcgacccttc ctggccatcc tgggcggagc taaagttgca
     1441 gacaagatcc agctcatcaa taatatgctg gacaaagtca atgagatgat tattggtggt
     1501 ggaatggctt ttaccttcct taaggtgctc aacaacatgg agattggcac ttctctgttt
     1561 gatgaagagg gagccaagat tgtcaaagac ctaatgtcca aagctgagaa gaatggtgtg
     1621 aagattacct tgcctgttga ctttgtcact gctgacaagt ttgatgagaa tgccaagact
     1681 ggccaagcca ctgtggcttc tggcatacct gctggctgga tgggcttgga ctgtggtcct
     1741 gaaagcagca agaagtatgc tgaggctgtc actcgggcta agcagattgt gtggaatggt
     1801 cctgtggggg tatttgaatg ggaagctttt gcccggggaa ccaaagctct catggatgag
     1861 gtggtgaaag ccacttctag gggctgcatc accatcatag gtggtggaga cactgccact
     1921 tgctgtgcca aatggaacac ggaggataaa gtcagccatg tgagcactgg gggtggtgcc
     1981 agtttggagc tcctggaagg taaagtcctt cctggggtgg atgctctcag caatattctc
     2041 gagcatgcat ctagaatggt gtctaagggc gaagagctga tcaaggaaaa tatgcgtatg
     2101 aaggtggtca tggaaggttc ggtcaacggc caccaattca aatgcacagg tgaaggagaa
     2161 ggcagaccgt acgagggagt gcaaaccatg aggatcaaag tcatcgaggg aggacccctg
     2221 ccatttgcct ttgacattct tgccacgtcg ttcatgtatg gcagccgtac ctttatcaag
     2281 tacccggccg acatccctga tttctttaaa cagtcctttc ctgagggttt tacttgggaa
     2341 agagttacga gatacgaaga tggtggagtc gtcaccgtca cgcaggacac cagccttgag
     2401 gatggcgagc tcgtctacaa cgtcaaggtc agaggggtaa actttccctc caatggtccc
     2461 gtgatgcaga agaagaccaa gggttgggag cctaatacag agatgatgta tccagcagat
     2521 ggtggtctga gaggatacac tgacatcgca ctgaaagttg atggtggtgg ccatctgcac
     2581 tgcaacttcg tgacaactta caggtcaaaa aagaccgtcg ggaacatcaa gatgcccggt
     2641 gtccatgccg ttgatcaccg cctggaaagg atcgaggaga gtgacaatga aacctacgta
     2701 gtgcaaagag aagtggcagt tgccaaatac agcaaccttg gtggtggcat ggacgagctg
     2761 tacaagtaga agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     2821 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     2881 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2941 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3001 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3061 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3121 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3181 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     3241 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3301 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     3361 gcctccccgc atcgataccg tcgacctcga cccgggcggc cgcttcgagc agacatgata
     3421 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     3481 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     3541 aacaacaaca attgcattca ttttatgttt caggttcagg gggagatgtg ggaggttttt
     3601 taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatcttcc tagagcatgg
     3661 ctacgtagat aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga
     3721 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     3781 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcattaa
     3841 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3901 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3961 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     4021 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4081 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4141 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4201 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4261 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4321 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4381 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4441 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4501 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4561 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4621 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4681 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4741 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4801 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4861 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4921 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4981 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5041 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5101 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5161 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5221 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5281 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5341 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5401 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5461 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5521 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5581 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5641 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5701 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5761 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     5821 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     5881 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     5941 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     6001 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     6061 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     6121 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     6181 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     6241 ccgccgcgct
//
