LOCUS       pOTTC1553_-_pAAV        6326 bp DNA     circular SYN 22-FEB-2024
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6326)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2024 from SnapGene 7.0.2
            https://www.snapgene.com
COMMENT     pOTTC1553 was constructed by Chris Richie at GEVVC/NIDA (CR1852_D).
            
            
            bakcbone was pOTTC407 - pAAV EF1a eGFP was created by Mari 
            Savolainen on 09/28/2012 (MS08).  Addgene 60058
            Described in Savolainen et al. (2014).  The beneficial effect of a 
            prolyl oligopeptidase inhibitor, KYP-2047, on alpha-synuclein 
            clearance and autophagy in A30P transgenic mouse.  Neurobiology of 
            Disease.  DOI: 10.1016/j.nbd.2014.04.003
FEATURES             Location/Qualifiers
     source          1..6326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4; direction: RIGHT"
     promoter        156..1419
                     /label=EF-1a promoter
                     /note="color: #408080"
     promoter        218..473
                     /label=EFS (EF1a short) promoter
                     /note="color: #8000ff"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
                     /note="color: #------"
     misc_feature    537..566
                     /label=+C in EF1a promoter
                     /note="GAATTACTTCCACTGGCTGCAGTACGTGAT -> GAATTACTTCCAC c 
                     TGGCTGCAGTACGTGAT
                     this is an existing -1nuc SNP in the EF1a promoter of many 
                     GEVVC vectors.
                     stemming from ""founder effect"", likely going back to KD's
                     optogenetic vectors.
                     
                     added to GEVVC Common Features on 10-02-23"
                     /note="color: #a6acb3"
     regulatory      1439..1448
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
                     /regulatory_class="other"
     CDS             1445..2428
                     /label=EGFP-KASH
                     /label=NucEnv-EGFP
                     /note="color: #0080c0; direction: RIGHT"
     CDS             1445..2161
                     /codon_start=1
                     /label=EGFP
                     /note="color: #0080c0"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             2180..2428
                     /label=KASH domain of Nesprin2
                     /note="/dnas_title=KASH domain of Nesprin2"
                     /note="/vntifkey=4"
                     /note="color: #0080c0; direction: RIGHT"
     misc_RNA        2443..3051
                     /label=WPRE
                     /note="color: #800080; direction: RIGHT"
     polyA_signal    3072..3552
                     /label=hGH polyA
                     /note="color: #8080ff"
     repeat_region   complement(3589..3729)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
                     /note="color: #ffe4c4; direction: LEFT"
     rep_origin      3804..4259
                     /label=M13 origin
                     /note="color: #808080"
     promoter        4541..4645
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: RIGHT"
     CDS             4646..5503
                     /label=AmpR
                     /note="color: #0080c0; direction: RIGHT"
     rep_origin      5601..6283
                     /label=ColE1 origin
                     /note="color: #808080"
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 caccggtcgc
     1441 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1501 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1561 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1621 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     1681 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     1741 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     1801 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     1861 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     1921 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     1981 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2041 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2101 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2161 gtccggactc agatctcgag aggaggagga ggagacagac agcaggatgc cccacctcga
     2221 cagccccggc agctcccagc cgagacgctc cttcctctca agggtgatca gggcagcgct
     2281 accgttgcag ctgcttctgc tgctgctgct gctcctggcc tgcctgctac ctgcctctga
     2341 agatgactac agctgcaccc aggccaacaa ctttgcccga tccttctacc ccatgctgcg
     2401 gtacaccaac gggccacctc ccacctagga attcgatatc aagcttatcg ataatcaacc
     2461 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2521 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2581 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2641 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2701 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2761 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2821 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     2881 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2941 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3001 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     3061 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     3121 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     3181 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     3241 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     3301 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     3361 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     3421 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     3481 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     3541 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     3601 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     3661 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     3721 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     3781 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3841 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3901 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3961 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4021 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4081 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4141 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4201 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4261 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4321 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     4381 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     4441 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     4501 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4561 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4621 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     4681 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4741 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     4801 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     4861 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     4921 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     4981 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5041 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5101 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5161 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5221 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5281 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5341 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     5401 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5461 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     5521 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5581 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5641 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5701 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5761 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5821 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5881 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5941 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6001 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6061 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6121 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6181 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6241 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6301 cttttgctgg ccttttgctc acatgt
//
