LOCUS       sequence-465749-        6371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of glutamate sensor with improved 
            sensitivity and slow deactivation kinetics (4s = slow); 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 6371)
  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 6371)
  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-iGluSnFR4s-NGR-WPRE
FEATURES             Location/Qualifiers
     source          1..6371
                     /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(996..1025)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1490..1511
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1614..1635)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2604..2663)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2660..2669
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2680..2713)
                     /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(2784..2817)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2859..3447
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2912..2932)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3330..3341)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3490..3611
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3501..3520)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3555..3574
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3773..3902
                     /label=AAV2 ITR
     rep_origin      complement(4139..4727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4219..4238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4898..5689,5690..5758))
                     /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     5521..5540
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5759..5863)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5889..6344)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6026..6047)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6238..6257
                     /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 tttcaacagg ggcccaacac cgtccacaga
      901 accaatgcca gtcccagtgg agccagacta catgcaaggg caggaagggc acctgatcct
      961 tctgcatctc ctgttccaga tgcaccacta cctgccagat cctcttctga gatgagtttt
     1021 tgttcgtcga cctgcagttt cagtgccttg tcattcggtt ctttgaacag tgctttcatt
     1081 tcgtctgaca gttcgaaatt catgaccagg gggttgttaa agttgtactc cagcttgtgc
     1141 cccaggatgt tgccgtcctc cttgaagtcg atgcccttta gctcgatgcg gttcaccagg
     1201 gtgtcgccct ctaacttcac ctcggcgcgg gtcttgtagg ttccgtcgtc cttgaagctg
     1261 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     1321 atgtggtcgg ggtagcgggc gaagcactgc acgccgtagc tcagggtggt cacgagggtg
     1381 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggttag cttgccgttg
     1441 gtggcatcgc cctcgccctc gccgcgcacg ctgaacttgt ggccgtttac gtcgccgtcc
     1501 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct catgctccct
     1561 ccggtaccgc ccttgtacag ctcgtccatg ccgagagagt tcccggtggc ggtcacgaac
     1621 tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagcac ggactgatag
     1681 ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg
     1741 tagtggtcgg cgagctgcac gctgccgtcc tccacgttgt ggtggatgaa gaagttcgcc
     1801 ttgatgccgt tcttctgctt gtcggcggtg atatagaccg tgtggttcac cagaattgga
     1861 tttttgaacc acttatcaaa ccatttttcc gcttcaccgg aggtctgcac ctgagcgatg
     1921 gtgtcatcca tcagcttttt gaactgcgga tcatctttac gcaacataca accgtaggcc
     1981 tcctgagact gcggcttgcc gacgatttcc cagtcgtctg gtttcttcgc tttcgcacgt
     2041 tcaccggcca gcagcgagtc atccatcata aaggcaacgg cacgaccgct ttccagggtg
     2101 cggaaagagt caccgtgatc tttggcgctg atgatgcgca tattcatttt ttgctcttca
     2161 ttcagtttgt tgagcaaaac ttcagaggta gtgccggaag tgacgactac ggctttgtct
     2221 ttcaggttgg caaaatcttt gatatcgcca ccctttttgg tcaacaggcg cgtaccgacc
     2281 acgaaaatag tgtcagagaa agccgcgaat ttttggcgtt cgacgttgtt ggtggtagaa
     2341 ccacattcaa aatcgtaagt gccgttttgc agtagtggaa tacggttttg tgaagtaatc
     2401 ggaatcagtt ttacctgcaa gtccggtttg ttgagtttct ttttcactgc ttcaacaatg
     2461 gcgttggagt attcctgcga gtaacccacc actttgtgct gactgtcttg ataagagaaa
     2521 ggcactgaag agtcacggtg accgacgaca atcacaccgt ttttggcgat tttgtccagc
     2581 gtgctgcctg cggcggatcc gtcaccagtg gaacctggaa cccagagcag cagtacccat
     2641 agcaggagtg tgtctgtctc catggtggcg gatcctagta taacttcgta taggatactt
     2701 tatacgaagt tatcattggg attcttccta ttttgatcca agcatcacca tcgaccctct
     2761 agtccagatc tcaccatcga cccataactt cgtatagcat acattatacg aagttatgtc
     2821 cctcgaagag gttcgaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa
     2881 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2941 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     3001 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     3061 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     3121 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     3181 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3241 tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc
     3301 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3361 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3421 ggatctccct ttgggccgcc tccccgcatc gataccgtcg acccgggcgg ccgcttcgag
     3481 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     3541 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     3601 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt
     3661 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc
     3721 ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc
     3781 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     3841 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     3901 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3961 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4021 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4081 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4141 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4201 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4261 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4321 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4381 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4441 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4501 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4561 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4621 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4681 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4741 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4801 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4861 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4921 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4981 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     5041 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5101 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5161 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5221 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5281 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5341 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5401 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5461 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5521 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5581 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5641 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5701 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5761 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5821 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5881 tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     5941 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     6001 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     6061 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     6121 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     6181 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     6241 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     6301 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     6361 gctgcgcaac t
//
