LOCUS Exported 7467 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION An AAV packaging vector that expresses hPREP under control of the EF1a promoter.. ACCESSION . VERSION . KEYWORDS pOTTC414 - pAAV EF1a hPREP 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 aggttcaaat 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 //