LOCUS       sequence-441103-        6919 bp    DNA     circular SYN 15-APR-2025
DEFINITION  AAV packaging construct for expression Sun1-sfGFP fusion Protein 
            under the human synapsin 1 gene promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6919)
  TITLE     Investigating the Enteric Nervous System of Parkinson's Model Mice
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6919)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-Hsyn-Sun1sfGFP
FEATURES             Location/Qualifiers
     source          1..6919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(196..215)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   513..653
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   513..642
                     /label=AAV2 ITR
     rep_origin      728..1183
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(815..834)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1025..1046
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1200..1219)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1319..1341
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1379..1397)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1465..1569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1570..1638,1639..2430)
                     /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(1788..1807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2601..3189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3090..3109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3251..3380
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        3425..3872
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(5556..5558,5559..6272)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             5556..6269
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Nager et al., 2011)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(5586..5614)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6209..6229
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    6296..6884
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6349..6369)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6767..6778)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    join(6916..6919,1..473)
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
ORIGIN
        1 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
       61 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
      121 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
      181 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
      241 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
      301 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
      361 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
      421 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
      481 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
      541 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
      601 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
      661 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
      721 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
      781 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
      841 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
      901 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
      961 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     1021 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     1081 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     1141 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     1201 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     1261 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     1321 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     1381 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     1441 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     1501 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     1561 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     1621 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     1681 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     1741 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     1801 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     1861 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     1921 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     1981 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2041 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2101 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2161 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2221 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     2281 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     2341 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     2401 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     2461 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     2521 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     2761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     2941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3241 gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg
     3301 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
     3361 ctccatcact aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg
     3421 tatgagtgca agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga
     3481 ccgaccccga cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat
     3541 cagagagggg gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac
     3601 cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag
     3661 gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg
     3721 tccgcgccgc cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg
     3781 ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca
     3841 gcggaggagt cgtgtcgtgc ctgagagcgc agtcgagaag gtaccaggga ggatccgcca
     3901 ccatgacctc cagggtgtac tccagagaca ggactctcaa accacggaag gcagcctcgg
     3961 gaaccttctg gtggctaggg agcggctggt accaatttgt tactttgatt tcttggctga
     4021 atgtctttct tcttaccagg tgccttcgaa atatttgcaa ggtttttgtc ttgctcctcc
     4081 cactcctact tttactaggt gctggtgtct ccctgtgggg ccagggaaac ttcttctcac
     4141 tcctaccagt gctgaactgg acggccatgc agccaacaca gagggtggac gattccaagg
     4201 gcatgcatag acctggccct cttcccccga gcccacctcc aaaggttgat cacaaggctt
     4261 cccagtggcc tcaggagagt gacatggggc agaaggtagc ttctttgagt gcgcagtgcc
     4321 acaaccatga tgagagactt gcagagctga cagtcctgct tcagaaacta cagatacggg
     4381 tagaccaagt ggatgacggc agggaagggc tgtcactgtg ggtcaagaat gtggttggac
     4441 agcacctgca ggagatgggc accatagaac cacctgatgc taagactgac ttcatgactt
     4501 tccaccatga ccatgaagtg cgtctctcca acttggaaga tgttcttaga aaactgacag
     4561 aaaaatctga ggctatccag aaggagctgg aagaaaccaa gctgaaagca ggcagcaggg
     4621 atgaagagca gcccctcctt gaccgtgtgc agcacctaga actggaactg aacctgttga
     4681 agtcacagct gtcagactgg cagcatctga agaccagctg tgagcaggct ggggcccgca
     4741 tccaggagac tgtgcagctc atgttctctg aggatcagca gggcggttcc ctcgagtggc
     4801 tattagagaa gctgtcttct cggttcgtga gcaaggatga gctgcaggtg ctcttacatg
     4861 accttgagct gaaactgctg cagaatatca cacaccacat caccgtgaca ggacaggccc
     4921 cgacatccga ggctattgtg tctgccgtga atcaggcagg gatttcagga atcacagaag
     4981 cgcaagcaca tatcattgtg aacaatgctc tgaagctgta ctcccaagac aagacgggga
     5041 tggtggactt tgctctggag tctggaggtg gcagcatcct aagcactcgg tgctctgaga
     5101 cctatgagac caagacggca ctgctgagcc tgtttggggt cccactgtgg tacttctcac
     5161 agtcacctcg agtggtgatc cagcccgaca tctacccagg gaattgctgg gcgttcaaag
     5221 gttcccaggg gtacctggtg gtgcggttgt ccatgaagat ctacccaacc acattcacca
     5281 tggaacacat tccaaagaca ctatcaccca ctggtaacat ctccagtgcc cccaaagact
     5341 ttgcagtcta tggactggaa acggagtatc aagaagaggg gcagcctctg ggacggttca
     5401 cctatgacca ggaaggagac tcactccaga tgttccacac actggaaaga cctgaccaag
     5461 ccttccagat agtagagctc cgggtcctgt ccaactgggg ccaccctgag tacacttgcc
     5521 tctaccggtt ccgagtccac ggagagccca tccagatgag caaaggagaa gaacttttca
     5581 ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg
     5641 tccgtggaga gggtgaaggt gatgctacaa acggaaaact cacccttaaa tttatttgca
     5701 ctactggaaa actacctgtt ccatggccaa cacttgtcac tactctgacc tatggtgttc
     5761 aatgcttttc ccgttatccg gatcacatga aacggcatga ctttttcaag agtgccatgc
     5821 ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga tgacgggacc tacaagacgc
     5881 gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg tatcgagtta aaaggtattg
     5941 attttaaaga agatggaaac attctcggac acaaactcga gtacaacttt aactcacaca
     6001 atgtatacat cacggcagac aaacaaaaga atggaatcaa agctaacttc aaaattcgcc
     6061 acaacgttga agatggttcc gttcaactag cagaccatta tcaacaaaat actccaattg
     6121 gcgatggccc tgtcctttta ccagacaacc attacctgtc gacacaatct gtcctttcga
     6181 aagatcccaa cgaaaagcgt gaccacatgg tccttcttga gtttgtaact gctgctggga
     6241 ttacacatgg catggatgag ctctacaaat aaaagcttct aggaagctta tcgataatca
     6301 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     6361 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     6421 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     6481 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     6541 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     6601 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     6661 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta
     6721 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     6781 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     6841 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct
     6901 gctcgagaga tctacgggt
//
