LOCUS       sequence-465724-        6062 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 6062)
  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 6062)
  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-iGluSnFR4s-NGR-WPRE
FEATURES             Location/Qualifiers
     source          1..6062
                     /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"
     misc_feature    2544..3132
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2597..2617)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3015..3026)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3135..3151)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3136..3152)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3181..3302
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3192..3211)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3246..3265
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3464..3593
                     /label=AAV2 ITR
     rep_origin      complement(3830..4418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3910..3929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4589..5380,5381..5449))
                     /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     5212..5231
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5450..5554)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5580..6035)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5717..5738)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5929..5948
                     /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 gatgactcgc 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 aggatctggc aggtagtggt gcatctggaa caggagatgc agaaggatca ggtgcccttc
     2461 ctgcccttgc atgtagtctg gctccactgg gactggcatt ggttctgtgg acggtgttgg
     2521 gcccctgtta aaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     2581 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2641 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2701 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     2761 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     2821 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     2881 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     2941 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     3001 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3061 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3121 ccgcctcccc gcatcgatac cgtcgacctc gacccgggcg gccgcttcga gcagacatga
     3181 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
     3241 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
     3301 ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggagatg tgggaggttt
     3361 tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatctt cctagagcat
     3421 ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc cctagtgatg
     3481 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     3541 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt
     3601 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3661 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3721 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3781 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3841 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3901 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3961 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4021 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4081 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4141 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4201 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4261 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4321 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4381 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4441 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4501 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4561 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4621 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4681 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4741 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4801 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4861 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4921 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4981 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5041 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5101 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5161 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5221 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5281 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5341 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5401 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5461 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     5521 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     5581 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5641 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5701 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5761 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5821 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5881 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     5941 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     6001 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     6061 ct
//
