LOCUS       sequence-488479-        7453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Large cargo capacity AAV that expresses SEP tagged AMPA receptor 
            subunit GluA1 Q582R mutant in neurons expressing Cre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7453)
  AUTHORS   Hong I, Kim J, Hainmueller T, Kim DW, Keijser J, Johnson RC, Park 
            SH, Limjunyawong N, Yang Z, Cheon D, Hwang T, Agarwal A, Cholvin T, 
            Krienen FM, McCarroll SA, Dong X, Leopold DA, Blackshaw S, Sprekeler
            H, Bergles DE, Bartos M, Brown SP, Huganir RL
  TITLE     Calcium-permeable AMPA receptors govern PV neuron feature 
            selectivity.
  JOURNAL   Nature. 2024 Nov;635(8038):398-405. doi: 10.1038/s41586-024-08027-2.
            Epub 2024 Oct 2.
  PUBMED    39358515
REFERENCE   2  (bases 1 to 7453)
  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"; doi: 
            "10.1038/s41586-024-08027-2"; journalName: "Nature"; date: 
            "2024-11"; volume: "635"; issue: "8038"; pages: "398-405"
COMMENT     Sequence Label: pAAV-hSyn1-DIO-SEP-GluA1R
FEATURES             Location/Qualifiers
     source          1..7453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        159..606
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(607..640)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(671..704)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3387..4097)
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(3427..3447)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     4042..4070
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    4182..4215
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    4246..4279
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(4336..4356)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4716..4856
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4716..4845
                     /label=AAV2 ITR
     rep_origin      4931..5386
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5018..5037)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5228..5249
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5403..5422)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5522..5544
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5582..5600)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5668..5772
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5773..5841,5842..6633)
                     /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(5991..6010)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6804..7392
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7293..7312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgctg cagagggccc tgcgtatgag tgcaagtggg ttttaggacc
      181 aggatgaggc ggggtggggg tgcctacctg acgaccgacc ccgacccact ggacaagcac
      241 ccaaccccca ttccccaaat tgcgcatccc ctatcagaga gggggagggg aaacaggatg
      301 cggcgaggcg cgtgcgcact gccagcttca gcaccgcgga cagtgccttc gcccccgcct
      361 ggcggcgcgc gccaccgccg cctcagcact gaaggcgcgc tgacgtcact cgccggtccc
      421 ccgcaaactc cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga
      481 ccgcaccacg cgaggcgcga gatagggggg cacgggcgcg accatctgcg ctgcggcgcc
      541 ggcgactcag cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga
      601 gcgcagataa cttcgtatag gatactttat acgaagttat gcagaatggt agctggattg
      661 tagctgctat ataacttcgt atagcataca ttatacgaag ttatctagag tcctgtggct
      721 cccaagggca tccctgaact gtgactcatg cagggaatgg attgcatgga cttggggaag
      781 tcctggctga ccacccggcc attctctcca ctgccgcctc ctccgctggc tcctgcccca
      841 ctgttccggg ggagggtcga tgtccgtatg gcttcattga tggattgctg tgggatcaaa
      901 cagaaaccct tcatccgctt cgactcgcta cgggatttgt agcagaactc gattaaggca
      961 accagcatgg ccagtcccag ccctccaatc aggatgtaga acacgcctgc cacattgctg
     1021 aggctcaaag cgctggtctt gtccttggag tcacctcccc cgctgccgca ctcgcccttg
     1081 tcgtaccacc atttgttttt caatttgtcc aaaagcccct gctcgttcag ttttaacact
     1141 gccaggttta ctggatttct cagggcggac cccttgggtg tcgcaatgcc atagcctttg
     1201 gaatccaagt tacctcccac tttcatggtg tcacagggct ttcgttgctc aatatactca
     1261 ttcatggtgg actccaggag gtaggcgtat ttgcctttag attttctcac tctgatcatg
     1321 ccttcctctg tggtccgaac aaacacggat ggttctgcag acttcatgta tgtccacatc
     1381 ttctcaaaca cagcgatttt agatctcctg aagaactcct tagtggagcc tgcttccaat
     1441 gtcccataag caatttccgt ctgctttgcc aggtcctctg cactctcaat gggagacacc
     1501 atcctctcca cagtcaggaa ggcagccagg ttggctgtgt acgaggagat gatgatcaaa
     1561 gtgaagaacc accagacgcc gccgacgatg cgtccggaca gggacctggg ggaaatgtca
     1621 catccttgcc gcatgaaggc ccccagggag aaccacaggc tgttgaatat gccaaactca
     1681 tttgactggt cactggttgt ctggtctcgt ccctcttcaa actcttcgct gtgccattcg
     1741 taggggctga aacggctgac caggaagagg acgacgctca ctccaatgta ggcaaacact
     1801 atacacatcc agatctcata ggccaaaggg tcaagaaagg agaagacacc tggcttggac
     1861 ttctgtggct tcttaatcat aatggagatt cccaaactca tgaatggctt ggagaagtcg
     1921 atgacttcct cccggaccaa ggttatggtc aagggagcca cagccacgtc tgctcttcca
     1981 tagaccagtt ctcccaccat gccattccaa gcctttgtgt cgggatcccg ggctccatat
     2041 ttgccgtcgc tgacaatctc aagtcggtag gagtagccca cgtgcttggc gatctctgca
     2101 gccagctcca cacagtagcc ctcatagcgg tcattgccct caaactggtt ggcattcttt
     2161 ttaagcatca cgtaaggatc ttcgaggata gtagtgacga tgtaggtcct attctggacg
     2221 cttgagttgt cccctccagc ctgagcgtcg gtggctgcgg ggacaaattt atcgtcttca
     2281 ttccagtaac caatctttcg gattccatca tgtttcattt cgatcacgtg gagggtgtaa
     2341 ttggtccggc gccctttctc gttgaactgc acatttcctg tcaaaccttc gaagcgcacc
     2401 tgctgcaggg ctctctggat gtcgatccct tgaccccagg gcacagctgg gttagccaga
     2461 cagtccccag cattcccccg gcgggatatg tcaatcctct gcctccgcag gctttggaag
     2521 gcctcagcca tcaccttgac accatcatag gtgagagcag aagtgtactt tggcctcttc
     2581 cagtccaccc tggtatggtc tcgggagtca cttgtcctcc attgctgcat gattctggct
     2641 gggatcgtgt ctgtgtagtt caccagctgg aaacctgtca cattggctcc gctctccttg
     2701 aacttattta agtcaatgtc catgaagccc agattggcga ggatgtagtg gtacccgatg
     2761 ccattctttt ctagcttcac gatctggccc aggatggcgt tgaggcgttc tgattcacag
     2821 tcaaccacca ccagcctctc ctttttcttc tccaggtcct gaaagagcat ccggtatcct
     2881 tcctcggtgg ttgtcagaat gttgacagcc gttacctgcc agttcttctc tgcggctgta
     2941 tccaagactc tctgcaggac tgacaggccc cggtcagcat cataaatgta gacaaaggtt
     3001 tgccacttgt aatggtcgat aatgctaatg agagcttcct gtagttccgg gcgtagctga
     3061 aggacaaatt gattggatgt gtcaacagga aaacttggag taatgaagca cacatggagg
     3121 gccccacaga aggaggtcag catgttgaca gtccttcgtt cataaaatcc aaagatggca
     3181 tagactcctt tggagaactg ggaacagaaa cggtaagtca tctcaaagct gtcgctgatg
     3241 ttcacaatat cgatctgggg aagcagcttg gggggctccg tgagttgtga caaagcaaac
     3301 ctaaaagccg catgttcctg tgattgttgg tttggaaata acccccctat ctggatattg
     3361 ttggggaaat taccggtaga tccacctttg tatagttcat ccatgccatg tgtaatccca
     3421 gcagctgtta caaactcaag aaggaccatg tggtctctct tttcgttggg atctttcgaa
     3481 agagtagaag ttgtaaacag gtaatggttg tctggtaaaa ggacgggccc atcgccaatt
     3541 ggagtatttt gttgataatg gtctgctagt tgaacgcctc catcttcaat gttgtgtcta
     3601 attttgaagt tagctttgat tccattcttt tgtttgtctg ccatgatgta cacctggtga
     3661 tcgttatagt tgtattccaa tttgtgtcca agaatgtttc catcttcttt aaaatcaata
     3721 ccttttaact cgattctatt aacaagggta tcaccttcaa acttgacttc agcacgtgtc
     3781 ttgtagttcc cgtcatcttt gaaaaatata gttctttcct gtacataacc ttcgggcatg
     3841 gcactcttga aaaagtcatg ccgtttcata tgatctgggt atcttgaaaa gcattgaaca
     3901 ccataagtta aagtagtgac aagtgttggc caaggaacag gtagttttcc agtagtgcaa
     3961 ataaatttaa gggtaagttt tccgtatgtt gcatcacctt caccctctcc actgacagaa
     4021 aatttgtgcc cattaacatc accatctaat tcaacaagaa ttgggacaac tccagtgaaa
     4081 agttcttctc ctttactgtt agaagagaca ccaaaaatca gtccccataa aacaggagaa
     4141 aggaggacag aaatatgcat aatcttttgc atttccaaga aataacttcg tataaagtat
     4201 cctatacgaa gttatttgcc ttaacccaga aattatcact gttatataac ttcgtataat
     4261 gtatgctata cgaagttatg ataatcaacc tctggattac aaaatttgtg aaagattgac
     4321 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     4381 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     4441 agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4501 gctgttgggc actgacaatt ccgtggtgtt tatttgtgaa atttgtgatg ctattgcttt
     4561 atttgtaacc atctagcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     4621 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     4681 agggggagat gtgggaggtt ttttaaagcg gccgcaggaa cccctagtga tggagttggc
     4741 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4801 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     4861 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     4921 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4981 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     5041 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     5101 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     5161 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     5221 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     5281 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     5341 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     5401 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5461 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5521 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5581 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5641 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5701 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5761 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5821 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5881 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5941 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6001 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6061 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6121 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6181 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6241 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6301 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6361 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6421 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6481 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6541 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6601 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6661 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6721 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6781 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6841 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6901 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6961 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7021 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7081 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7141 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7201 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7261 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7321 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7381 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7441 tttgctcaca tgt
//
