LOCUS       sequence-481669-        7363 bp    DNA     circular SYN 01-AUG-2025
DEFINITION  Large cargo capacity AAV that expresses SEP tagged AMPA receptor 
            subunit GluA2 in neurons expressing Cre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7363)
  TITLE     Calcium-permeable AMPA receptors govern PV neuron feature 
            selectivity
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 1, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-hSyn1-DIO-SEP-GluA2
FEATURES             Location/Qualifiers
     source          1..7363
                     /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(3297..4007)
                     /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(3337..3357)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     3952..3980
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    4092..4125
                     /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    4156..4189
                     /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(4246..4266)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4626..4766
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4626..4755
                     /label=AAV2 ITR
     rep_origin      4841..5296
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4928..4947)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5138..5159
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5313..5332)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5432..5454
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5492..5510)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5578..5682
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5683..5751,5752..6543)
                     /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(5901..5920)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6714..7302
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7203..7222
                     /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 ttatctaaat tttaacactc
      721 tcgatgccat atacgttgta accttcctta taagttgcaa aattctggga attctgcgag
      781 gaagatgggt taatattctg tggattcttt gccaccttca ttcgtttcgc ctcggccctt
      841 gacttgtaac agaactcaat caaagccacc agcattgcca aaccaaggcc cccgacaagg
      901 atgtagaata ctccagcaac gttgctcaga ctgagggcac tggtcttttc cttacttccc
      961 gagtccttgg ctccacattc acctttatcg taccaccatt tgtttttcag cttgtctaag
     1021 acgccttgct cactgagttt caatactgca agatttactg gggttcctaa tgaggatcct
     1081 ttaggtgtgg cgatgccgta gcctttggaa tccaagtttc ctcccacttt catggtgtca
     1141 caaggcttcc tctgctcgat gtactcgttc attgtggact ccagcaagta ggcatacttt
     1201 cctttggatt tccggactct ggctactcct tctgcggtag tcctcacaaa cacagagggc
     1261 tctgcactcc tcatataagt ccacatttta tcaaacactg cgattttaga tctcctgaaa
     1321 aactctttag tggagccaga gtctaatgtt ccataagcaa tttctgtttg cttagacaga
     1381 tcctcagcac tttcgatggg agacaccatc ctctctacag tcaggaaggc agctaagtta
     1441 gccgtgtagg aggagattat gatcagggta aagaaccacc acacacctcc aacaatgcgc
     1501 ccagagaggg atcttggcga aatatcgcat ccttgccgca taaaggcacc caaggaaaac
     1561 cagagactat taaaaatccc aaattcatta gttgattcac tactttgtgt ttctcttcca
     1621 tcttcaaatt cctcagtgtg ccactcgtag gggctaaatc tgctgaccag gaataaaact
     1681 acactgaccc caatgtaggc aaacacaatg cacatccaga tctcataggc taaaggatca
     1741 agaaaggaaa acactcctgg tttggacttc tgaggcttct tgatcatgat agagattcca
     1801 agactcatga agggcttgga gaagtcaatc acctcttctc tcacgagagt gatagttaat
     1861 ggagcaattg caatgtcagc tttcccgtag acaagctctc caaccatacc attccaaatt
     1921 ttggtgtcgg catccctggc cccatacttg ccatccccaa caatagtcag cttgtacttg
     1981 aacccacagt gtttggcaat ttctgcagct aagtcaacac agtagccctc gtaacgctca
     2041 ttcccttcaa gcatttcatg atttttcttc atcataacat atggagattc caatattgtg
     2101 gtgaccacca cagtcttgtt ttcaagccca gacgtgtcat ttcctgatgg gagctcagtt
     2161 agggtgacaa ccattttatc cacttcactc cagtacccaa tcttccgggg tccatttgtt
     2221 ttgagctcca tgatgttaat tgtgtagttt attcgttttc cattctggtc aaactttata
     2281 tttccagaga ggccttcaac ttgaacctgc ttgagggccc tttctatttc gaccccttgt
     2341 ccccagggca cagctgggtt ggccaaacaa tcccctgcat ttcctctccg ggatatttca
     2401 atcctctgct tccgaaggtt acggaatgct tcagtcatca cttggacagc atcatacgtc
     2461 agggccgaag tatacttaat tgtcgctgtg tgtgctccag ggtattcttt ctcttccagt
     2521 gttgaccatc tttctataaa tttagacacc agggaatcat cgtagtctac aatctgaaat
     2581 ccagagacat ttgctcctcc aaactgaatt ttcagcaggt ccccatcagt gaatcccaga
     2641 tttgcaatga tataatggta ccctttaaca tgttttccaa tggtaataac ctggtccaca
     2701 atgtcattta ctttatccct ttcacagtcc aggattacac gccgttcttt ttttaactcc
     2761 agatcttgaa agagcgatct gtaggtctca tctttcttgt cattgttgat gttccccaca
     2821 ttgatagcag tcacctgcca cttcttctct gcagcagaat ccagaacagc ttgcagtgtt
     2881 gataagcctc tgtcactgtc atagaggtat gcgaacttgt cccattggta gtactcaatc
     2941 aagctaagga gtgctccttt gaggtcaggt cgcatctgga tgacaaatgg atgtgtgcca
     3001 tctgttggga agctaggtgt gatgaaggac acatggagtg tcccacagaa tgatgtgatg
     3061 gtatttacag acttcttgtc ataaaatcca aaaattgcgt agactcctct tgaaaactgg
     3121 gagcagaaag cattggtgac tgcgaaactg ttggctacct ccaaattgtc gatatggggt
     3181 gtcagtctga actccgaagt ggaaaactga accatcccta cccgaaatgc actgtattct
     3241 tgatcagcgc cccttggaaa tagcccccct atctgtatgc taccggtaga tccacctttg
     3301 tatagttcat ccatgccatg tgtaatccca gcagctgtta caaactcaag aaggaccatg
     3361 tggtctctct tttcgttggg atctttcgaa agagtagaag ttgtaaacag gtaatggttg
     3421 tctggtaaaa ggacgggccc atcgccaatt ggagtatttt gttgataatg gtctgctagt
     3481 tgaacgcctc catcttcaat gttgtgtcta attttgaagt tagctttgat tccattcttt
     3541 tgtttgtctg ccatgatgta cacctggtga tcgttatagt tgtattccaa tttgtgtcca
     3601 agaatgtttc catcttcttt aaaatcaata ccttttaact cgattctatt aacaagggta
     3661 tcaccttcaa acttgacttc agcacgtgtc ttgtagttcc cgtcatcttt gaaaaatata
     3721 gttctttcct gtacataacc ttcgggcatg gcactcttga aaaagtcatg ccgtttcata
     3781 tgatctgggt atcttgaaaa gcattgaaca ccataagtta aagtagtgac aagtgttggc
     3841 caaggaacag gtagttttcc agtagtgcaa ataaatttaa gggtaagttt tccgtatgtt
     3901 gcatcacctt caccctctcc actgacagaa aatttgtgcc cattaacatc accatctaat
     3961 tcaacaagaa ttgggacaac tccagtgaaa agttcttctc ctttactgtt agaagagaca
     4021 ccaaaaatca gtccccataa aacaggagaa aggaggacag aaatatgcat aatcttttgc
     4081 atttccaaga aataacttcg tataaagtat cctatacgaa gttatttgcc ttaacccaga
     4141 aattatcact gttatataac ttcgtataat gtatgctata cgaagttatg ataatcaacc
     4201 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     4261 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     4321 cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac tcatcgccgc
     4381 ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt
     4441 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc atctagcttt atttgtgaaa
     4501 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     4561 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt ttttaaagcg
     4621 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4681 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4741 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     4801 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     4861 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     4921 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4981 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     5041 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     5101 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     5161 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     5221 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     5281 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     5341 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     5401 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     5461 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     5521 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     5581 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5641 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5701 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5761 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5821 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5881 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5941 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     6001 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     6061 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     6121 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     6181 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     6241 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     6301 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     6361 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     6421 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     6481 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     6541 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     6601 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6661 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6721 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6781 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6841 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6901 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6961 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     7021 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     7081 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     7141 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     7201 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     7261 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     7321 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
