LOCUS       sequence-385091-        5103 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  knockdown alpha-synuclein in rat neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5103)
  AUTHORS   Zharikov AD, Cannon JR, Tapias V, Bai Q, Horowitz MP, Shah V, El 
            Ayadi A, Hastings TG, Greenamyre JT, Burton EA
  TITLE     shRNA targeting alpha-synuclein prevents neurodegeneration in a 
            Parkinson's disease model.
  JOURNAL   J Clin Invest. 2015 Jul 1;125(7):2721-35. doi: 10.1172/JCI64502. 
            Epub 2015 Jun 15.
  PUBMED    26075822
REFERENCE   2  (bases 1 to 5103)
  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"; doi: "10.1172/JCI64502"; 
            journalName: "J Clin Invest"; date: "2015-07-1- 1"; volume: "125"; 
            issue: "7"; pages: "2721-35"
COMMENT     Sequence Label: p-AAV-sh[SNCA]
FEATURES             Location/Qualifiers
     source          1..5103
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   55..184
                     /label=AAV2 ITR
     promoter        254..502
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     254..274
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     425..444
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     enhancer        628..931
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        932..1135
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1086..1106
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1174..1183
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1180..1182,1183..1185,1186..1899)
                     /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(1180..1182,1183..1185,1186..1896)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1225..1246)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1486..1505)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1833..1854
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1924..2045
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1935..1954)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1989..2008
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2207..2336
                     /label=AAV2 ITR
     primer_bind     complement(2355..2372)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2355..2371)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2364..2386)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2513..2968
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2600..2619)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2810..2831
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2994..3098
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3099..3167,3168..3959)
                     /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(3317..3336)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4130..4718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4619..4638
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5006..5027
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5042..5047,5048..5065,5066..5072)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5080..5096
                     /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     join(5085..5103,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca gatttaatta aggccttaat taggctgcgc
       61 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
      121 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
      181 tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag
      241 atctgggcag gaagagggcc tatttcccat gattccttca tatttgcata tacgatacaa
      301 ggctgttaga gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa
      361 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
      421 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      481 cttgtggaaa ggacgaggat ccccgcagtg aggcttatga aatttcaaga gaatttcata
      541 agcctcactg cttttttgga aaaattctag ttattaatag taatcaatta cggggtcatt
      601 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      661 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
      721 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
      781 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
      841 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
      901 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
      961 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     1021 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     1081 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     1141 agtgaaccgt cagatccgct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
     1201 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1261 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1321 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1381 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1441 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1501 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1561 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1621 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1681 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1741 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     1801 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1861 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgctt cgagcagaca
     1921 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     1981 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2041 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     2101 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag
     2161 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
     2221 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     2281 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt
     2341 aattaaccta attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     2401 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     2461 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     2521 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2581 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2641 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2701 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2761 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2821 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2881 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2941 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     3001 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3061 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     3121 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     3181 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     3241 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     3301 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     3361 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     3421 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3481 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3541 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3601 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3661 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3721 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3781 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3841 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3901 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3961 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     4021 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     4081 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     4141 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     4201 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     4261 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     4321 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     4381 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4441 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4501 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4561 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4621 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4681 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4741 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4801 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4861 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     4921 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4981 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     5041 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     5101 aca
//
