LOCUS       sequence-378605-        7971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of AMPA GluA1 with N terminal super ecliptic phluorin 
            (SEP) fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7971)
  AUTHORS   Kim D, Park P, Li X, Wong Campos JD, Tian H, Moult EM, Grimm JB, 
            Lavis L, Cohen AE
  TITLE     Mapping memories: pulse-chase labeling reveals AMPA receptor 
            dynamics during memory formation.
  JOURNAL   bioRxiv. 2023 May 29:2023.05.26.541296. doi: 
            10.1101/2023.05.26.541296. Preprint.
  PUBMED    37292614
REFERENCE   2  (bases 1 to 7971)
  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: "bioRxiv. 
            2023 May 29:2023.05.26.541296. doi: 10.1101/2023.05.26.541296. 
            Preprint."
COMMENT     Sequence Label: TL026_AAV_hSyn:SEP-GluA1
FEATURES             Location/Qualifiers
     source          1..7971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        151..598
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(636..669)
                     /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(720..753)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(3469..4179)
                     /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(3509..3529)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     4124..4152
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     protein_bind    4294..4327
                     /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    4378..4411
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4436..5024
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4489..4509)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4907..4918)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5078..5097)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5132..5151
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5210..5350
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5210..5339
                     /label=AAV2 ITR
     rep_origin      5425..5880
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5512..5531)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5722..5743
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5897..5916)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6016..6038
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6076..6094)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6162..6266
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6267..6335,6336..7127)
                     /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(6485..6504)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7298..7886
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7787..7806
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(7948..7971,1..106)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
       61 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgcacgcgt
      121 gtgtctagac tgcagagggc cctgcgtatg agtgcaagtg ggttttagga ccaggatgag
      181 gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc
      241 cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg
      301 cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc
      361 gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac
      421 tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca
      481 cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc
      541 agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc
      601 gagaaggtac cggatcctct agagtcgact ccggaataac ttcgtatagg atactttata
      661 cgaagttatg cagaatggta gctggattgt agctgctatt agcaatatga aacctcttaa
      721 taacttcgta tagcatacat tatacgaagt tatggcgcgc ccttacaatc ctgtggctcc
      781 caagggcatc cctgaactgt gactcatgca gggaatggat tgcatggact tggggaagtc
      841 ctggctgacc acccggccat tctctccact gccgcctcct ccgctggctc ctgccccact
      901 gttccggggg agggtcgatg tccgtatggc ttcattgatg gattgctgtg ggatcaaaca
      961 gaaacccttc atccgcttcg actcgctacg ggatttgtag cagaactcga ttaaggcaac
     1021 cagcatggcc agtcccagcc ctccaatcag gatgtagaac acgcctgcca cattgctgag
     1081 gctcaaagcg ctggtcttgt ccttggagtc acctcccccg ctgccgcact cgcccttgtc
     1141 gtaccaccat ttgtttttca atttgtccaa aagcccctgc tcgttcagtt ttaacactgc
     1201 caggtttact ggatttctca gggcggaccc cttgggtgtc gcaatgccat agcctttgga
     1261 atccaagtta cctcccactt tcatggtgtc acagggcttt cgttgctcaa tatactcatt
     1321 catggtggac tccaggaggt aggcgtattt gcctttagat tttctcactc tgatcatgcc
     1381 ttcctctgtg gtccgaacaa acacggatgg ttctgcagac ttcatgtatg tccacatctt
     1441 ctcaaacaca gcgattttag atctcctgaa gaactcctta gtggagcctg cttccaatgt
     1501 cccataagca atttccgtct gctttgccag gtcctctgca ctctcaatgg gagacaccat
     1561 cctctccaca gtcaggaagg cagccaggtt ggctgtgtac gaggagatga tgatcaaagt
     1621 gaagaaccac cagacgccgc cgacgatgcg tccggacagg gacctggggg aaatgtcaca
     1681 tccttgctgc atgaaggccc ccagggagaa ccacaggctg ttgaatatgc caaactcatt
     1741 tgactggtca ctggttgtct ggtctcgtcc ctcttcaaac tcttcgctgt gccattcgta
     1801 ggggctgaaa cggctgacca ggaagaggac gacgctcact ccaatgtagg caaacactat
     1861 acacatccag atctcatagg ccaaagggtc aagaaaggag aagacacctg gcttggactt
     1921 ctgtggcttc ttaatcataa tggagattcc caaactcatg aatggcttgg agaagtcgat
     1981 gacttcctcc cggaccaagg ttatggtcaa gggagccaca gccacgtctg ctcttccata
     2041 gaccagttct cccaccatgc cattccaagc ctttgtgtcg ggatcccggg ctccatattt
     2101 gccgtcgctg acaatctcaa gtcggtagga gtagcccacg tgcttggcga tctctgcagc
     2161 cagctccaca cagtagccct catagcggtc attgccctca aactggttgg cattcttttt
     2221 aagcatcacg taaggatctt cgaggatagt agtgacgatg taggtcctat tctggacgct
     2281 tgagttgtcc cctccagcct gagcgtcggt ggctgcgggg acaaatttat cgtcttcatt
     2341 ccagtaacca atctttcgga ttccatcatg tttcatttcg atcacgtgga gggtgtaatt
     2401 ggtccggcgc cctttctcgt tgaactgcac atttcctgtc aaaccttcga agcgcacctg
     2461 ctgcagggct ctctggatgt cgatcccttg accccagggc acagctgggt tagccagaca
     2521 gtccccagca ttcccccggc gggatatgtc aatcctctgc ctccgcaggc tttggaaggc
     2581 ctcagccatc accttgacac catcataggt gagagcagaa gtgtactttg gcctcttcca
     2641 gtccaccctg gtatggtctc gggagtcact tgtcctccat tgctgcatga ttctggctgg
     2701 gatcgtgtct gtgtagttca ccagctggaa acctgtcaca ttggctccgc tctccttgaa
     2761 cttatttaag tcaatgtcca tgaagcccag attggcgagg atgtagtggt acccgatgcc
     2821 attcttttct agcttcacga tctggcccag gatggcgttg aggcgttctg attcacagtc
     2881 aaccaccacc agcctctcct ttttcttctc caggtcctga aagagcatcc ggtatccttc
     2941 ctcggtggtt gtcagaatgt tgacagccgt tacctgccag ttcttctctg cggctgtatc
     3001 caagactctc tgcaggactg acaggccccg gtcagcatca taaatgtaga caaaggtttg
     3061 ccacttgtaa tggtcgataa tgctaatgag agcttcctgt agttccgggc gtagctgaag
     3121 gacaaattga ttggatgtgt caacaggaaa acttggagta atgaagcaca catggagggc
     3181 cccacagaag gaggtcagca tgttgacagt ccttcgttca taaaatccaa agatggcata
     3241 gactcctttg gagaactggg aacagaaacg gtaagtcatc tcaaagctgt cgctgatgtt
     3301 cacaatatcg atctggggaa gcagcttggg gggctccgtg agttgtgaca aagcaaacct
     3361 aaaagccgca tgttcctgtg attgttggtt tggaaataac ccccctatct ggatattgtt
     3421 ggggaagcgc gaacccccgc cacctccgcc ccctccacct ccacgcgttt tgtatagttc
     3481 atccatgcca tgtgtaatcc cagcagctgt tacaaactca agaaggacca tgtggtctct
     3541 cttttcgttg ggatctttcg aaagagtaga agttgtaaac aggtaatggt tgtctggtaa
     3601 aaggacgggc ccatcgccaa ttggagtatt ttgttgataa tggtctgcta gttgaacgcc
     3661 tccatcttca atgttgtgtc taattttgaa gttagctttg attccattct tttgtttgtc
     3721 tgccatgatg tacacctggt gatcgttata gttgtattcc aatttgtgtc caagaatgtt
     3781 tccatcttct ttaaaatcaa taccttttaa ctcgattcta ttaacaaggg tatcaccttc
     3841 aaacttgact tcagcacgtg tcttgtagtt cccgtcatct ttgaaaaata tagttctttc
     3901 ctgtacataa ccttcgggca tggcactctt gaaaaagtca tgccgtttca tatgatctgg
     3961 gtatcttgaa aagcattgaa caccataagt taaagtagtg acaagtgttg gccaaggaac
     4021 aggtagtttt ccagtagtgc aaataaattt aagggtaagt tttccgtatg ttgcatcacc
     4081 ttcaccctct ccactgacag aaaatttgtg cccattaaca tcaccatcta attcaacaag
     4141 aattgggaca actccagtga aaagttcttc tcctttacta cgcgttcctc caccccctcc
     4201 cccaccgcct ccacctcgcg cattggcacc cacaaccgca cctagaaaac cggtgcagaa
     4261 aaaggcaaag atgtacggca tggtggcgct agcataactt cgtataaagt atcctatacg
     4321 aagttatttg ccttaaccca gaaattatca ctgttattct ttagaatggt gcaaagaata
     4381 acttcgtata atgtatgcta tacgaagtta tgaattcgat atcaagctta tcgataatca
     4441 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     4501 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4561 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4621 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     4681 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     4741 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4801 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta
     4861 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4921 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4981 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     5041 cgggcggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     5101 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     5161 aaccattata agctgcaata aactaaccac gtgcggaccg agcggccgca ggaaccccta
     5221 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     5281 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     5341 tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     5401 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     5461 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg
     5521 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     5581 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     5641 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     5701 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta
     5761 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa
     5821 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     5881 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     5941 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     6001 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     6061 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     6121 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     6181 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     6241 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     6301 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     6361 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     6421 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     6481 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     6541 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     6601 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     6661 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     6721 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     6781 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     6841 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     6901 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     6961 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     7021 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     7081 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     7141 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     7201 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     7261 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     7321 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     7381 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     7441 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     7501 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     7561 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     7621 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     7681 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7741 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7801 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7861 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7921 ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc t
//
