LOCUS       sequence-465719-        6092 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 6092)
  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 6092)
  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-iGluSnFR4f-PDGFR-WPRE
FEATURES             Location/Qualifiers
     source          1..6092
                     /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)"
     regulatory      735..744
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     741..800
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1769..1790
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1893..1914)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2379..2408
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2412..2558
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2574..3162
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2627..2647)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3045..3056)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3165..3181)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3166..3182)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3211..3332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3222..3241)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3276..3295
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3494..3623
                     /label=AAV2 ITR
     rep_origin      complement(3860..4448)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3940..3959)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4619..5410,5411..5479))
                     /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     5242..5261
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5480..5584)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5610..6065)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5747..5768)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5959..5978
                     /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 acccgccacc atggagacag acacactcct gctatgggta ctgctgctct
      781 gggttccagg ttccactggt gacggatccg ccgcaggcag cacgctggac aaaatcgcca
      841 aaaacggtgt gattgtcgtc ggtcaccgtg actcttcagt gcctttctct tatcaagaca
      901 gtcagcacaa agtggtgggt tactcgcagg aatactccaa cgccattgtt gaagcagtga
      961 aaaagaaact caacaaaccg gacttgcagg taaaactgat tccgattact tcacaaaacc
     1021 gtattccact actgcaaaac ggcacttacg attttgaatg tggttctacc accaacaacg
     1081 tcgaacgcca aaaattcgcg gctttctctg acactatttt cgtggtcggt acgcgcctgt
     1141 tgaccaaaaa gggtggcgat atcaaagatt ttgccaacct gaaagacaaa gccgtagtcg
     1201 tcacttccgg cactacctct gaagttttgc tcaacaaact gaatgaagag caaaaaatga
     1261 atatgcgcat catcagcgcc aaagatcacg gtgactcttt ccgcaccctg gaaagcggtc
     1321 gtgccgttgc ctttatgatg gatgacgtac tgctggccgg tgaacgtgcg aaagcgaaga
     1381 aaccagacga ctgggaaatc gtcggcaagc cgcagtctca ggaggcctac ggttgtatgt
     1441 tgcgtaaaga tgatccgcag ttcaaaaagc tgatggatga caccatcgct caggtgcaga
     1501 cctccggtga agcggaaaaa tggtttgata agtggttcaa aaatccaatt ctggtgaacc
     1561 acacggtcta tatcaccgcc gacaagcaga agaacggcat caaggcgaac ttcttcatcc
     1621 accacaacgt ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1681 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctatcag tccgtgctga
     1741 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccaccg
     1801 ggaactctct cggcatggac gagctgtaca agggcggtac cggagggagc atgagcaagg
     1861 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     1921 gccacaagtt cagcgtgcgc ggcgagggcg agggcgatgc caccaacggc aagctaaccc
     1981 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2041 tgagctacgg cgtgcagtgc ttcgcccgct accccgacca catgaagcag cacgacttct
     2101 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcagcttc aaggacgacg
     2161 gaacctacaa gacccgcgcc gaggtgaagt tagagggcga caccctggtg aaccgcatcg
     2221 agctaaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2281 actttaacaa ccccctggtc atgaatttcg aactgtcaga cgaaatgaaa gcactgttca
     2341 aagaaccgaa tgacaaggca ctgaaactgc aggtcgacga acaaaaactc atctcagaag
     2401 aggatctgaa tgctgtgggc caggacacgc aggaggtcat cgtggtgcca cactccttgc
     2461 cctttaaggt ggtggtgatc tcagccatcc tggccctggt ggtgctcacc atcatctccc
     2521 ttatcatcct catcatgctt tggcagaaga agccacgtta aaagcttatc gataatcaac
     2581 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2641 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2701 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2761 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2821 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2881 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     2941 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     3001 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     3061 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     3121 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc
     3181 gacccgggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     3241 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3301 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     3361 ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     3421 gtaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta
     3481 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3541 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3601 tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     3661 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     3721 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     3781 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     3841 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     3901 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     3961 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4021 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4081 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4141 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4201 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     4261 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     4321 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     4381 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     4441 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     4501 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     4561 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     4621 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     4681 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     4741 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     4801 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     4861 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     4921 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     4981 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5041 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5101 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5161 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     5221 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     5281 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     5341 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     5401 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     5461 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     5521 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     5581 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     5641 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     5701 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     5761 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     5821 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     5881 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     5941 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6001 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     6061 cgcgtcccat tcgccattca ggctgcgcaa ct
//
