LOCUS       sequence-465739-        6286 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 6286)
  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 6286)
  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-GFAP-iGluSnFR4s-PDGFR-WPRE
FEATURES             Location/Qualifiers
     source          1..6286
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR
     regulatory      929..938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     935..994
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1963..1984
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2087..2108)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2573..2602
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2606..2752
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2768..3356
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2821..2841)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3239..3250)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3359..3375)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3360..3376)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3405..3526
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3416..3435)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3470..3489
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3688..3817
                     /label=AAV2 ITR
     rep_origin      complement(4054..4642)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4134..4153)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4813..5604,5605..5673))
                     /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"
     promoter        complement(5674..5778)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5804..6259)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5941..5962)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6153..6172
                     /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 atccaacata
      241 tcctggtgtg gagtagggga cgctgctctg acagaggctc gggggcctga gctggctctg
      301 tgagctgggg aggaggcaga cagccaggcc tttgtctgca agcagacctg gcagcattgg
      361 gctggccgcc ccccagggcc tcctcttcat gcccagtgaa tgactcacct tggcacagac
      421 acaatgttcg gggtgggcac agtgcctgct tcccgccgca ccccagcccc cctcaaatgc
      481 cttccgagaa gcccattgag cagggggctt gcattgcacc ccagcctgac agcctggcat
      541 cttgggataa aagcagcaca gccccctagg ggctgccctt gctgtgtggc gccaccggcg
      601 gtggagaaca aggctctatt cagcctgtgc ccaggaaagg ggatcagggg atgcccaggc
      661 atggacagtg ggtggcaggg ggggagagga gggctgtctg cttcccagaa gtccaaggac
      721 acaaatgggt gaggggagag ctctccccat agctgggctg cggcccaacc ccaccccctc
      781 aggctatgcc agggggtgtt gccaggggca cccgggcatc gccagtctag cccactcctt
      841 cataaagccc tcgcatccca ggagcgagca gagccagagc aggttggaga ggagacgcat
      901 cacctccgct gctcgcgggg atccacccgc caccatggag acagacacac tcctgctatg
      961 ggtactgctg ctctgggttc caggttccac tggtgacgga tccgccgcag gcagcacgct
     1021 ggacaaaatc gccaaaaacg gtgtgattgt cgtcggtcac cgtgactctt cagtgccttt
     1081 ctcttatcaa gacagtcagc acaaagtggt gggttactcg caggaatact ccaacgccat
     1141 tgttgaagca gtgaaaaaga aactcaacaa accggacttg caggtaaaac tgattccgat
     1201 tacttcacaa aaccgtattc cactactgca aaacggcact tacgattttg aatgtggttc
     1261 taccaccaac aacgtcgaac gccaaaaatt cgcggctttc tctgacacta ttttcgtggt
     1321 cggtacgcgc ctgttgacca aaaagggtgg cgatatcaaa gattttgcca acctgaaaga
     1381 caaagccgta gtcgtcactt ccggcactac ctctgaagtt ttgctcaaca aactgaatga
     1441 agagcaaaaa atgaatatgc gcatcatcag cgccaaagat cacggtgact ctttccgcac
     1501 cctggaaagc ggtcgtgccg ttgcctttat gatggatgac tcgctgctgg ccggtgaacg
     1561 tgcgaaagcg aagaaaccag acgactggga aatcgtcggc aagccgcagt ctcaggaggc
     1621 ctacggttgt atgttgcgta aagatgatcc gcagttcaaa aagctgatgg atgacaccat
     1681 cgctcaggtg cagacctccg gtgaagcgga aaaatggttt gataagtggt tcaaaaatcc
     1741 aattctggtg aaccacacgg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     1801 gaacttcttc atccaccaca acgtggagga cggcagcgtg cagctcgccg accactacca
     1861 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcta
     1921 tcagtccgtg ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     1981 cgtgaccgcc accgggaact ctctcggcat ggacgagctg tacaagggcg gtaccggagg
     2041 gagcatgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2101 cggcgacgta aacggccaca agttcagcgt gcgcggcgag ggcgagggcg atgccaccaa
     2161 cggcaagcta accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2221 cctcgtgacc accctgagct acggcgtgca gtgcttcgcc cgctaccccg accacatgaa
     2281 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatcag
     2341 cttcaaggac gacggaacct acaagacccg cgccgaggtg aagttagagg gcgacaccct
     2401 ggtgaaccgc atcgagctaa agggcatcga cttcaaggag gacggcaaca tcctggggca
     2461 caagctggag tacaacttta acaaccccct ggtcatgaat ttcgaactgt cagacgaaat
     2521 gaaagcactg ttcaaagaac cgaatgacaa ggcactgaaa ctgcaggtcg acgaacaaaa
     2581 actcatctca gaagaggatc tgaatgctgt gggccaggac acgcaggagg tcatcgtggt
     2641 gccacactcc ttgcccttta aggtggtggt gatctcagcc atcctggccc tggtggtgct
     2701 caccatcatc tcccttatca tcctcatcat gctttggcag aagaagccac gttaaaagct
     2761 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     2821 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     2881 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
     2941 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
     3001 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
     3061 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     3121 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
     3181 gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
     3241 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     3301 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     3361 ataccgtcga cctcgacccg ggcggccgct tcgagcagac atgataagat acattgatga
     3421 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     3481 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     3541 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
     3601 cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt
     3661 agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct
     3721 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3781 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa tcggccaacg
     3841 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3901 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3961 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4021 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4081 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4141 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4201 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4261 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4321 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4381 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4441 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4501 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4561 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4621 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4681 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4741 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4801 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4861 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4921 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4981 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5041 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5101 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5161 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5221 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5281 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5341 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5401 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5461 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5521 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5581 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5641 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5701 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5761 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     5821 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     5881 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     5941 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     6001 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     6061 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     6121 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     6181 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     6241 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaact
//
