LOCUS       sequence-465720-        6401 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of glutamate sensor with improved 
            sensitivity and fast deactivation kinetics (4f = fast); NGR vector 
            matches or outperforms PDGFR in all conditions tested..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6401)
  AUTHORS   Aggarwal A, Negrean A, Chen Y, Iyer R, Reep D, Liu A, Palutla A, Xie
            ME, MacLennan BJ, Hagihara KM, Kinsey LW, Sun JL, Yao P, Zheng J, 
            Tsang A, Tsegaye G, Zhang Y, Patel RH, Arthur BJ, Hiblot J, Leippe 
            P, Tarnawski M, Marvin JS, Vevea JD, Turaga SC, Tebo AG, Carandini 
            M, Rossi LF, Kleinfeld D, Konnerth A, Svoboda K, Turner GC, Hasseman
            J, Podgorski K
  TITLE     Glutamate indicators with increased sensitivity and tailored 
            deactivation rates
  JOURNAL   bioRxiv 2025.03.20.643984
REFERENCE   2  (bases 1 to 6401)
  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 
            2025.03.20.643984"
COMMENT     Sequence Label: pAAV-syn-flex-iGluSnFR4f-PDGFR-WPRE
FEATURES             Location/Qualifiers
     source          1..6401
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR
     promoter        256..703
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    733..766
                     /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    829..862
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(876..1022)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1026..1055)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1520..1541
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1644..1665)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2634..2693)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2690..2699
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2710..2743)
                     /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(2814..2847)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2889..3477
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2942..2962)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3360..3371)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3520..3641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3531..3550)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3585..3604
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3803..3932
                     /label=AAV2 ITR
     rep_origin      complement(4169..4757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4249..4268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4928..5719,5720..5788))
                     /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     5551..5570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5789..5893)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5919..6374)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6056..6077)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6268..6287
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc
       61 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
      121 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt
      181 taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag atctctgcag
      241 agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc
      301 ctacctgacg accgaccccg acccactgga caagcaccca acccccattc cccaaattgc
      361 gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc
      421 agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct
      481 cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca
      541 ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat
      601 aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc
      661 tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagtcgaatt caagctgcta
      721 gcaaggatcc ccataacttc gtataaagta tcctatacga agttatatca aaataggaag
      781 accaatgctt caccatcgac ccgaattgcc aagcatcacc atcgacccat aacttcgtat
      841 aatgtatgct atacgaagtt atactagagc tttcaacgtg gcttcttctg ccaaagcatg
      901 atgaggatga taagggagat gatggtgagc accaccaggg ccaggatggc tgagatcacc
      961 accaccttaa agggcaagga gtgtggcacc acgatgacct cctgcgtgtc ctggcccaca
     1021 gcattcagat cctcttctga gatgagtttt tgttcgtcga cctgcagttt cagtgccttg
     1081 tcattcggtt ctttgaacag tgctttcatt tcgtctgaca gttcgaaatt catgaccagg
     1141 gggttgttaa agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
     1201 atgcccttta gctcgatgcg gttcaccagg gtgtcgccct ctaacttcac ctcggcgcgg
     1261 gtcttgtagg ttccgtcgtc cttgaagctg atggtgcgct cctggacgta gccttcgggc
     1321 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc
     1381 acgccgtagc tcagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg
     1441 cagatgaact tcagggttag cttgccgttg gtggcatcgc cctcgccctc gccgcgcacg
     1501 ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg
     1561 aacagctcct cgcccttgct catgctccct ccggtaccgc ccttgtacag ctcgtccatg
     1621 ccgagagagt tcccggtggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
     1681 ttggggtctt tgctcagcac ggactgatag ctcaggtagt ggttgtcggg cagcagcacg
     1741 gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc
     1801 tccacgttgt ggtggatgaa gaagttcgcc ttgatgccgt tcttctgctt gtcggcggtg
     1861 atatagaccg tgtggttcac cagaattgga tttttgaacc acttatcaaa ccatttttcc
     1921 gcttcaccgg aggtctgcac ctgagcgatg gtgtcatcca tcagcttttt gaactgcgga
     1981 tcatctttac gcaacataca accgtaggcc tcctgagact gcggcttgcc gacgatttcc
     2041 cagtcgtctg gtttcttcgc tttcgcacgt tcaccggcca gcagtacgtc atccatcata
     2101 aaggcaacgg cacgaccgct ttccagggtg cggaaagagt caccgtgatc tttggcgctg
     2161 atgatgcgca tattcatttt ttgctcttca ttcagtttgt tgagcaaaac ttcagaggta
     2221 gtgccggaag tgacgactac ggctttgtct ttcaggttgg caaaatcttt gatatcgcca
     2281 ccctttttgg tcaacaggcg cgtaccgacc acgaaaatag tgtcagagaa agccgcgaat
     2341 ttttggcgtt cgacgttgtt ggtggtagaa ccacattcaa aatcgtaagt gccgttttgc
     2401 agtagtggaa tacggttttg tgaagtaatc ggaatcagtt ttacctgcaa gtccggtttg
     2461 ttgagtttct ttttcactgc ttcaacaatg gcgttggagt attcctgcga gtaacccacc
     2521 actttgtgct gactgtcttg ataagagaaa ggcactgaag agtcacggtg accgacgaca
     2581 atcacaccgt ttttggcgat tttgtccagc gtgctgcctg cggcggatcc gtcaccagtg
     2641 gaacctggaa cccagagcag cagtacccat agcaggagtg tgtctgtctc catggtggcg
     2701 gatcctagta taacttcgta taggatactt tatacgaagt tatcattggg attcttccta
     2761 ttttgatcca agcatcacca tcgaccctct agtccagatc tcaccatcga cccataactt
     2821 cgtatagcat acattatacg aagttatgtc cctcgaagag gttcgaattc gatatcaagc
     2881 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2941 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3001 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     3061 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3121 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     3181 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3241 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     3301 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3361 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3421 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3481 gataccgtcg acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     3541 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3601 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3661 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
     3721 acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat
     3781 ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg
     3841 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3901 cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg
     3961 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4021 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4081 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4141 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4201 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4261 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4321 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4381 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4441 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4501 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4561 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4621 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4681 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4741 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4801 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4861 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4921 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4981 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5041 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5101 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5161 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5221 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5281 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5341 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5401 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5461 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5521 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5581 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5641 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5701 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5761 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5821 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5881 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     5941 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     6001 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     6061 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     6121 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     6181 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     6241 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     6301 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     6361 gctacagggc gcgtcccatt cgccattcag gctgcgcaac t
//
