LOCUS       Exported                7467 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  An AAV packaging vector that expresses catalytically-inactive hPREP 
            under control of the EF1a promoter..
ACCESSION   .
VERSION     .
KEYWORDS    pOTTC446 - pAAV EF1a hPREP(S554A)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7467)
  AUTHORS   Savolainen MH, Richie CT, Harvey BK, Mannisto PT, Maguire-Zeiss KA, 
            Myohanen TT
  TITLE     The beneficial effect of a prolyl oligopeptidase inhibitor, 
            KYP-2047, on alpha-synuclein clearance and autophagy in A30P 
            transgenic mouse.
  JOURNAL   Neurobiol Dis. 2014 Aug;68:1-15. doi: 10.1016/j.nbd.2014.04.003. 
            Epub 2014 Apr 16.
  PUBMED    24746855
REFERENCE   2  (bases 1 to 7467)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7467
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     misc_feature    3594..4182
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3647..3667)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4065..4076)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4214..4690
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4413..4432)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4730..4870
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4730..4859
                     /label=AAV2 ITR
     rep_origin      4945..5400
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5032..5051)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5242..5263
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5417..5436)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5536..5558
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5596..5614)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5682..5786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5787..6647
                     /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(6005..6024)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6818..7406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7307..7326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gtaccggatc cggccatgct
     1441 gtcccttcag taccccgacg tgtaccgcga cgagaccgcc gtacaggatt atcatggtca
     1501 taaaatttgt gacccttacg cctggcttga agaccccgac agtgaacaga ctaaggcctt
     1561 tgtggaggcc cagaataaga ttactgtgcc atttcttgag cagtgtccca tcagaggttt
     1621 atacaaagag agaatgactg aactatatga ttatcccaag tatagttgcc acttcaagaa
     1681 aggaaaacgg tatttttatt tttacaatac aggtttgcag aaccagcgag tattatatgt
     1741 acaggattcc ttagagggtg aggccagagt gttcctggac cccaacatac tgtctgacga
     1801 tggcacagtg gcactccgag gttacgcgtt cagcgaagat ggtgaatatt ttgcctatgg
     1861 tctgagtgcc agtggctcag actgggtgac aatcaagttc atgaaagttg atggtgccaa
     1921 agagcttcca gatgtgcttg aaagagtcaa gttcagctgt atggcctgga cccatgatgg
     1981 gaagggaatg ttctacaact cataccctca acaggatgga aaaagtgatg gcacagagac
     2041 atctaccaat ctccaccaaa agctctacta ccatgtcttg ggaaccgatc agtcagaaga
     2101 tattttgtgt gctgagtttc ctgatgaacc taaatggatg ggtggagctg agttatctga
     2161 tgatggccgc tatgtcttgt tatcaataag ggaaggatgt gatccagtaa accgactctg
     2221 gtactgtgac ctacagcagg aatccagtgg catcgcggga atcctgaagt gggtaaaact
     2281 gattgacaac tttgaagggg aatatgacta cgtgaccaat gaggggacgg tgttcacatt
     2341 caagacgaat cgccagtctc ccaactatcg cgtgatcaac attgacttca gggatcctga
     2401 agagtctaag tggaaagtac ttgttcctga gcatgagaaa gatgtcttag aatggatagc
     2461 ttgtgtcagg tccaacttct tggtcttatg ctacctccat gacgtcaaga acattctgca
     2521 gctccatgac ctgactactg gtgctctcct taagaccttt ccgctcgatg tcggcagcat
     2581 tgtagggtac agcggtcaga agaaggacac tgaaatcttc tatcagttta cttccttttt
     2641 atctccaggt atcatttatc actgtgatct taccaaagag gagctggagc caagagtttt
     2701 ccgagaggtg accgtaaaag gaattgatgc ttctgattac cagacagtcc agattttcta
     2761 ccctagcaag gatggtacga agattccaat gttcattgtg cataaaaaag gcataaaatt
     2821 ggatggctct catccagctt tcttatatgg ctatggcggc ttcaacatat ccatcacacc
     2881 caactacagt gtttccaggc ttatttttgt gagacacatg ggtggtatcc tggcagtggc
     2941 caacatcaga ggaggtggcg aatatggaga gacgtggcat aaaggtggta tcttggccaa
     3001 caaacaaaac tgctttgatg actttcagtg tgctgctgag tatctgatca aggaaggtta
     3061 cacatctccc aagaggctga ctattaatgg aggcgccaat ggaggcctct tagtggctgc
     3121 ttgtgcaaat cagagacctg acctctttgg ttgtgttatt gcccaagttg gagtaatgga
     3181 catgctgaag tttcataaat ataccatcgg ccatgcttgg accactgatt atgggtgctc
     3241 ggacagcaaa caacactttg aatggcttgt caaatactct ccattgcata atgtgaagtt
     3301 accagaagca gatgacatcc agtacccgtc catgctgctc ctcactgctg accatgatga
     3361 ccgcgtggtc ccgcttcact ccctgaagtt cattgccacc cttcagtaca tcgtgggccg
     3421 cagcaggaag caaagcaacc ccctgcttat ccacgtggac accaaggcgg gccacggggc
     3481 ggggaagccc acagccaaag tgatagagga agtctcagac atgtttgcgt tcatcgcgcg
     3541 gtgcctgaac gtcgactgga ttccataaca ctcgagatat caagcttatc gataatcaac
     3601 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3661 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3721 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     3781 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     3841 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     3901 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3961 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctatg
     4021 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     4081 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     4141 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgagcgctgc
     4201 tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc tggccctgga
     4261 agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca tcattttgtc
     4321 tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga gcaaggggca
     4381 agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt
     4441 ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc tcctgcctca
     4501 gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat ttttgttttt
     4561 ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct aatctcaggt
     4621 gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca ctgctccctt
     4681 ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca ggaaccccta
     4741 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     4801 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     4861 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4921 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     4981 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     5041 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5101 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5161 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5221 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     5281 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     5341 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     5401 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     5461 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     5521 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     5581 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     5641 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     5701 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     5761 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     5821 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     5881 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     5941 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6001 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6061 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6121 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6181 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6241 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6301 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6361 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6421 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6481 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     6541 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     6601 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     6661 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6721 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6781 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6841 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6901 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6961 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7021 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7081 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7141 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7201 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7261 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7321 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7381 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7441 ccttttgctg gccttttgct cacatgt
//
