LOCUS       sequence-208748-        8042 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral overexpression of synuclein alpha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8042)
  AUTHORS   Ho PW, Leung CT, Liu H, Pang SY, Lam CS, Xian J, Li L, Kung MH, 
            Ramsden DB, Ho SL
  TITLE     Age-dependent accumulation of oligomeric SNCA/alpha-synuclein from 
            impaired degradation in mutant LRRK2 knockin mouse model of 
            Parkinson disease: role for therapeutic activation of 
            chaperone-mediated autophagy (CMA).
  JOURNAL   Autophagy. 2019 Apr 14:1-24. doi: 10.1080/15548627.2019.1603545.
  PUBMED    30983487
REFERENCE   2  (bases 1 to 8042)
  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: "Autophagy. 
            2019 Apr 14:1-24. doi: 10.1080/15548627.2019.1603545."
COMMENT     Sequence Label: pSIN-hSNCA-NE
FEATURES             Location/Qualifiers
     source          1..8042
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1299..1532
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1717..1761
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2048..2165
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2294..3472
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2525..3463
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3420..3440
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      3480..3489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(4042..4616,4617..4619,4620..4628)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4209..4226)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4436..4455
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4648..5247
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4648..4667)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5144..5164
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5450..5683
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     primer_bind     complement(5834..5851)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5840..6064
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     regulatory      6107..6116
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        6160..6264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6265..6333,6334..7125)
                     /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(6483..6502)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7296..7884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7785..7804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgc tgatcttcag acctggagga ggagatatga gggacaattg gagaagtgaa
     1201 ttatataaat ataaagtagt aaaaattgaa ccattaggag tagcacccac caaggcaaag
     1261 agaagagtgg tgcagagaga aaaaagagca gtgggaatag gagctttgtt ccttgggttc
     1321 ttgggagcag caggaagcac tatgggcgca gcgtcaatga cgctgacggt acaggccaga
     1381 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
     1441 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
     1501 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
     1561 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
     1621 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
     1681 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
     1741 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
     1801 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
     1861 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
     1921 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
     1981 gacagatcca ttcgattagt gaacggatct cgacggtatc gccacaaatg gcagtattca
     2041 tccacaattt taaaagaaag ggggggattg gggggtacag tgcaggggaa agaatagtag
     2101 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa
     2161 attttcgggt ttattacagg gacagcagag atccactttg gatcgataag ctttgcaaag
     2221 atggataaag ttttaaacag agaggaatct ttgcagctaa tggaccttct aggtcttgaa
     2281 aggagtggga attggctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
     2341 cgagaagttg gggggagggg tcggcaattg aaccggtgcc tagagaaggt ggcgcggggt
     2401 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
     2461 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
     2521 acaggtaagt gccgtgtgtg gttcccgcgg gcctggcctc tttacgggtt atggcccttg
     2581 cgtgccttga attacttcca cctggctgca gtacgtgatt cttgatcccg agcttcgggt
     2641 tggaagtggg tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg
     2701 agttgaggcc tggcctgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc
     2761 ctgtctcgct gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac
     2821 gctttttttc tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc
     2881 ggtttttggg gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag
     2941 gcggggcctg cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc
     3001 tgctctggtg cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc
     3061 ccggtcggca ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag
     3121 ctcaaaatgg aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa
     3181 aagggccttt ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc
     3241 caggcacctc gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg
     3301 gttttatgcg atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg
     3361 gcacttgatg taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct
     3421 caagcctcag acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggaattcg
     3481 ccaccatgga tgtattcatg aaaggacttt caaaggccaa ggagggagtt gtggctgctg
     3541 ctgagaaaac caaacagggt gtggcagaag cagcaggaaa gacaaaagag ggtgttctct
     3601 atgtaggctc caaaaccaag gagggagtgg tgcatggtgt ggcaacagtg gctgagaaga
     3661 ccaaagagca agtgacaaat gttggaggag cagtggtgac gggtgtgaca gcagtagccc
     3721 agaagacagt ggagggagca gggagcattg cagcagccac tggctttgtc aaaaaggacc
     3781 agttgggcaa gaatgaagaa ggagccccac aggaagggat tctggaagat atgcctgtgg
     3841 atcctgacaa tgaggcttat gaaatgcctt ctgaggaagg gtatcaagac tacgaacctg
     3901 aagccaccaa agaaaacccg cgtagcaacc aggaagaaag ctatgatgat aacgaaagct
     3961 aagctagcta agtttaaaca atcactagtt cgaaggatcc gcatgcatct agggcggcca
     4021 attccgcccc tctccccccc ccccctctcc ctcccccccc cctaacgtta ctggccgaag
     4081 ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc
     4141 ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg
     4201 tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc
     4261 tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc
     4321 cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa
     4381 ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct
     4441 ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg
     4501 atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg
     4561 tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataagcttg
     4621 ccacaaccca caaggagacg accttccatg accgagtaca agcccacggt gcgcctcgcc
     4681 acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc cgactacccc
     4741 gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga gctgcaagaa
     4801 ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc
     4861 gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc
     4921 ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc
     4981 ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggcgtctcg
     5041 cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc
     5101 gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac
     5161 gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg
     5221 tgcatgaccc gcaagcccgg tgcctgataa taggcggccg ctcgagacct agaaaaacat
     5281 ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg gctagaagca
     5341 caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag accaatgact
     5401 tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta
     5461 attcactccc aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac
     5521 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa
     5581 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag
     5641 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat
     5701 cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga
     5761 cattataata gatttagcag gaattgaact aggagtggag cacacaggca aagttctaga
     5821 gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
     5881 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
     5941 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
     6001 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
     6061 atggcttctg aggcggaaag aaccagctgg gggcgcgccc ctcgaggccg ccatggtcat
     6121 agctgtttga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6181 tttttctaaa tacattcaaa tatgtatccg ctcacgagac aataaccctg ataaatgctt
     6241 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6301 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6361 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6421 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6481 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     6541 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6601 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6661 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6721 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6781 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     6841 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6901 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6961 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     7021 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     7081 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     7141 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7201 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7261 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7321 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7381 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7441 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7501 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7561 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7621 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7681 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7741 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7801 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7861 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttctggcct
     7921 tttgcacgga ccgccactgc caattacctg tggtttcatt tactctaaac ctgtgattcc
     7981 tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac aaacttagta
     8041 gt
//
