LOCUS       sequence-378842-        4390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector expressing thew GFP reporter gene and human sgHTT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4390)
  AUTHORS   Duarte F, Ramosaj M, Hasanovic E, Regio S, Sipion M, Rey M, Deglon N
  TITLE     Semi-automated workflows to quantify AAV transduction in various 
            brain areas and predict gene editing outcome for neurological 
            disorders.
  JOURNAL   Mol Ther Methods Clin Dev. 2023 Mar 27;29:254-270. doi: 
            10.1016/j.omtm.2023.03.013. eCollection 2023 Jun 8.
  PUBMED    37090478
REFERENCE   2  (bases 1 to 4390)
  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"; journalName: "Mol Ther 
            Methods Clin Dev."; date: "2023-03-27"; volume: "29:254-270. doi"; 
            pages: " 10.1016/j.omtm.2023.03.013. eCollection 2023 Jun 8"
COMMENT     Sequence Label: pAAV2ss-EFS-GFP-synPolyA-U6-sgHTT1
FEATURES             Location/Qualifiers
     source          1..4390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   16..145
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        206..417
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      426..435
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(432..434,435..437,438..1148)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(432..434,435..437,438..1148)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(477..498)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(738..757)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1085..1106
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1211..1259
                     /note="synthetic polyadenylation signal"
     promoter        1287..1535
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1287..1307
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     1458..1477
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     repeat_region   1668..1808
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1668..1797
                     /label=AAV2 ITR
     rep_origin      1883..2338
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1970..1989)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2180..2201
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2355..2374)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2474..2496
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2534..2552)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2620..2724
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2725..2793,2794..3585)
                     /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(2943..2962)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3756..4344
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4245..4264
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
       61 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      121 gccaactcca tcactagggg ttcctgcggc ctctagaaag cttaggtctt gaaaggagtg
      181 ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
      241 ttggggggag gggtcggcaa ttgatccggt gcctagagaa ggtggcgcgg ggtaaactgg
      301 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
      361 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggac
      421 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      481 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      541 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      601 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
      661 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      721 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      781 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
      841 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
      901 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
      961 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1021 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1081 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1141 tgtacaagtc cggactcaga tctcgactag ctagtagcta gctagctagt cgagctcaag
     1201 cttcgaattc aataaaagat ctttattttc attagatctg tgtgttggtt ttttgtgtgc
     1261 ggccgctggg atccaagctt ccatgggagg gcctatttcc catgattcct tcatatttgc
     1321 atatacgata caaggctgtt agagagataa ttggaattaa tttgactgta aacacaaaga
     1381 tattagtaca aaatacgtga cgtagaaagt aataatttct tgggtagttt gcagttttaa
     1441 aattatgttt taaaatggac tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct
     1501 tggctttata tatcttgtgg aaaggacgaa acaccgaccc tggaaaagct gatgagtttc
     1561 agagctatgc tggaaacagc atagcaagtt gaaataaggc tagtccgtta tcaacttgaa
     1621 aaagtggcac cgagtcggtg ctttttttca tagccatggg cggccgcagg aacccctagt
     1681 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     1741 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     1801 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     1861 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1921 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     1981 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2041 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     2101 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2161 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     2221 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     2281 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     2341 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     2401 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     2461 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     2521 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     2581 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2641 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2701 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2761 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2821 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2881 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2941 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3001 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3061 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3121 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3181 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3241 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3301 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3361 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3421 tctggagccg gtgagcgtgg aagccgcggt atcattgcag cactggggcc agatggtaag
     3481 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3541 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3601 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3661 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3721 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3781 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3841 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3901 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3961 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4021 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4081 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4141 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4201 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4261 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4321 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4381 ttttgctggc
//
