LOCUS       sequence-385081-        5102 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  control shRNA vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5102)
  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 5102)
  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[control]
FEATURES             Location/Qualifiers
     source          1..5102
                     /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        627..930
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        931..1134
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1085..1105
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1173..1182
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1179..1181,1182..1184,1185..1898)
                     /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(1179..1181,1182..1184,1185..1895)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1224..1245)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1485..1504)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1832..1853
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1923..2044
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1934..1953)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1988..2007
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2206..2335
                     /label=AAV2 ITR
     primer_bind     complement(2354..2371)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2354..2370)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2363..2385)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2512..2967
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2599..2618)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2809..2830
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2993..3097
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3098..3166,3167..3958)
                     /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(3316..3335)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4129..4717
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4618..4637
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5005..5026
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5041..5046,5047..5064,5065..5071)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5079..5095
                     /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(5084..5102,1..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 ccccaattct ccgaacgtgt cacgtttcaa gagaacgtga
      541 cacgttcgga gaattttttg gaattctagt tattaatagt aatcaattac ggggtcatta
      601 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      661 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      721 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      781 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      841 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      901 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      961 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     1021 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     1081 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
     1141 gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
     1201 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1261 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1321 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1381 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1441 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     1501 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     1561 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1621 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1681 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1741 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1801 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1861 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcttc gagcagacat
     1921 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     1981 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     2041 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     2101 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc
     2161 atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga
     2221 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     2281 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagcctta
     2341 attaacctaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     2401 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     2461 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct
     2521 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2581 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2641 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2701 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2761 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2821 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2881 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2941 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac
     3001 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     3061 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     3121 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     3181 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     3241 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     3301 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     3361 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3421 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3481 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3541 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3601 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3661 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3721 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3781 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3841 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3901 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3961 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     4021 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4081 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4141 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4201 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4261 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4321 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4381 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4441 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4501 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4561 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4621 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4681 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4741 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4801 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4861 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     4921 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     4981 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     5041 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     5101 ca
//
