LOCUS       sequence-465732-        6502 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 6502)
  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 6502)
  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-iGluSnFR4s-NGR-WPRE
FEATURES             Location/Qualifiers
     source          1..6502
                     /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"
     misc_feature    2990..3578
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3043..3063)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3461..3472)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3621..3742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3632..3651)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3686..3705
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3904..4033
                     /label=AAV2 ITR
     rep_origin      complement(4270..4858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4350..4369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5029..5820,5821..5889))
                     /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     5652..5671
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5890..5994)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6020..6475)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6157..6178)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6369..6388
                     /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 actcgctgct
     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 tctggcaggt agtggtgcat ctggaacagg
     2881 agatgcagaa ggatcaggtg cccttcctgc ccttgcatgt agtctggctc cactgggact
     2941 ggcattggtt ctgtggacgg tgttgggccc ctgttaaaag cttatcgata atcaacctct
     3001 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3061 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3121 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3181 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3241 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3301 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3361 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     3421 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3481 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3541 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacccgggcg
     3601 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3661 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3721 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3781 gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     3841 taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     3901 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3961 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4021 agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4081 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4141 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4201 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4261 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4321 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4381 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4441 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4501 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4561 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4621 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4681 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4741 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4801 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4861 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4921 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4981 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5041 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5101 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5161 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5221 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5281 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5341 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5401 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5461 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5521 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5581 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5641 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5701 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5761 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5821 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5881 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5941 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6001 tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     6061 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     6121 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     6181 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     6241 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     6301 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     6361 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     6421 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat
     6481 tcgccattca ggctgcgcaa ct
//
