LOCUS       sequence-465729-        6532 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 6532)
  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 6532)
  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-CAG-iGluSnFR4f-PDGFR-WPRE
FEATURES             Location/Qualifiers
     source          1..6532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR
     promoter        485..762
                     /label=chicken beta-actin promoter
     primer_bind     1041..1063
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1125..1144
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1181..1190
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1187..1246
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     2215..2236
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2339..2360)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2825..2854
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2858..3004
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    3020..3608
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3073..3093)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3491..3502)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3651..3772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3662..3681)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3716..3735
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3934..4063
                     /label=AAV2 ITR
     rep_origin      complement(4300..4888)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4380..4399)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5059..5850,5851..5919))
                     /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     5682..5701
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5920..6024)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6050..6505)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6187..6208)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6399..6418
                     /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 agtaccattg
      241 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      301 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      361 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      421 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      481 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
      541 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg
      601 ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga
      661 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
      721 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
      781 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
      841 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
      901 ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg
      961 gagggccctt tgtgcggggg gagcggctcg gggctgtccg cggggggacg gctgccttcg
     1021 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     1081 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     1141 tgtgctgtct catcattttg gcaaagaatt ggatccaccc gccaccatgg agacagacac
     1201 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgacg gatccgccgc
     1261 aggcagcacg ctggacaaaa tcgccaaaaa cggtgtgatt gtcgtcggtc accgtgactc
     1321 ttcagtgcct ttctcttatc aagacagtca gcacaaagtg gtgggttact cgcaggaata
     1381 ctccaacgcc attgttgaag cagtgaaaaa gaaactcaac aaaccggact tgcaggtaaa
     1441 actgattccg attacttcac aaaaccgtat tccactactg caaaacggca cttacgattt
     1501 tgaatgtggt tctaccacca acaacgtcga acgccaaaaa ttcgcggctt tctctgacac
     1561 tattttcgtg gtcggtacgc gcctgttgac caaaaagggt ggcgatatca aagattttgc
     1621 caacctgaaa gacaaagccg tagtcgtcac ttccggcact acctctgaag ttttgctcaa
     1681 caaactgaat gaagagcaaa aaatgaatat gcgcatcatc agcgccaaag atcacggtga
     1741 ctctttccgc accctggaaa gcggtcgtgc cgttgccttt atgatggatg acgtactgct
     1801 ggccggtgaa cgtgcgaaag cgaagaaacc agacgactgg gaaatcgtcg gcaagccgca
     1861 gtctcaggag gcctacggtt gtatgttgcg taaagatgat ccgcagttca aaaagctgat
     1921 ggatgacacc atcgctcagg tgcagacctc cggtgaagcg gaaaaatggt ttgataagtg
     1981 gttcaaaaat ccaattctgg tgaaccacac ggtctatatc accgccgaca agcagaagaa
     2041 cggcatcaag gcgaacttct tcatccacca caacgtggag gacggcagcg tgcagctcgc
     2101 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2161 ctacctgagc tatcagtccg tgctgagcaa agaccccaac gagaagcgcg atcacatggt
     2221 cctgctggag ttcgtgaccg ccaccgggaa ctctctcggc atggacgagc tgtacaaggg
     2281 cggtaccgga gggagcatga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     2341 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgcgcggcg agggcgaggg
     2401 cgatgccacc aacggcaagc taaccctgaa gttcatctgc accaccggca agctgcccgt
     2461 gccctggccc accctcgtga ccaccctgag ctacggcgtg cagtgcttcg cccgctaccc
     2521 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     2581 gcgcaccatc agcttcaagg acgacggaac ctacaagacc cgcgccgagg tgaagttaga
     2641 gggcgacacc ctggtgaacc gcatcgagct aaagggcatc gacttcaagg aggacggcaa
     2701 catcctgggg cacaagctgg agtacaactt taacaacccc ctggtcatga atttcgaact
     2761 gtcagacgaa atgaaagcac tgttcaaaga accgaatgac aaggcactga aactgcaggt
     2821 cgacgaacaa aaactcatct cagaagagga tctgaatgct gtgggccagg acacgcagga
     2881 ggtcatcgtg gtgccacact ccttgccctt taaggtggtg gtgatctcag ccatcctggc
     2941 cctggtggtg ctcaccatca tctcccttat catcctcatc atgctttggc agaagaagcc
     3001 acgttaaaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     3061 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     3121 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3181 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3241 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3301 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3361 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     3421 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     3481 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3541 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3601 ctccccgcat cgataccgtc gacccgggcg gccgcttcga gcagacatga taagatacat
     3661 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     3721 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     3781 caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa
     3841 gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt cctagagcat ggctacgtag
     3901 ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca
     3961 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     4021 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg
     4081 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4141 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4201 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4261 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4321 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4381 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4441 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4501 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4561 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4621 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4681 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4741 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4801 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4861 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4921 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4981 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5041 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5101 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5161 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5221 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5281 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5341 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5401 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     5461 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5521 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5581 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5641 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5701 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5761 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5821 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5881 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5941 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6001 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
     6061 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     6121 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     6181 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     6241 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     6301 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     6361 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     6421 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     6481 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ct
//
