LOCUS       sequence-478670-        4781 bp    DNA     circular SYN 15-JUL-2025
DEFINITION  Large capacity AAV that expresses eGFP in neurons expressing Cre 
            recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4781)
  TITLE     Calcium-permeable AMPA receptors govern PV neuron feature 
            selectivity
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4781)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 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            "Unpublished"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-hSyn1-DIO-eGFP
FEATURES             Location/Qualifiers
     source          1..4781
                     /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(join(775..1488,1489..1491,1492..1494))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(778..1488,1489..1491,1492..1494))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(820..841)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1169..1188
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1428..1449
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1491..1500
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1510..1543
                     /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    1574..1607
                     /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(1664..1684)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2044..2184
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2044..2173
                     /label=AAV2 ITR
     rep_origin      2259..2714
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2346..2365)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2556..2577
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2731..2750)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2850..2872
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2910..2928)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2996..3100
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3101..3169,3170..3961)
                     /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(3319..3338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4132..4720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4621..4640
                     /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 attcctactt
      781 gtacagctcg tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat
      841 gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt
      901 gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag
      961 ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt
     1021 ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc
     1081 caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt
     1141 gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat
     1201 ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat
     1261 gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg
     1321 ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt
     1381 ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag
     1441 ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatggtggc
     1501 gacctcgaga taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa
     1561 ttatcactgt tatataactt cgtataatgt atgctatacg aagttatgat aatcaacctc
     1621 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     1681 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     1741 ttttctcctc cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct
     1801 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttta
     1861 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat ctagctttat ttgtgaaatt
     1921 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     1981 aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcggc
     2041 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     2101 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     2161 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     2221 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     2281 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     2341 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2401 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2461 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2521 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2581 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     2641 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2701 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2761 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2821 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2881 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     2941 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3001 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3061 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3121 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3181 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3241 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3301 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3361 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3421 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3481 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3541 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3601 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3661 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3721 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3781 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3841 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3901 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3961 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4021 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4081 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4141 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4201 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4261 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4321 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4381 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4441 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4501 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4561 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4621 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4681 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4741 tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
