LOCUS       sequence-213318-        6324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Weak affinity glutamate sensor ** A newer version of this sensor is 
            available. Please see iGluSnFR3 plasmids at 
            https://www.addgene.org/browse/article/28220233/ **.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6324)
  AUTHORS   Marvin JS, Scholl B, Wilson DE, Podgorski K, Kazemipour A, Muller 
            JA, Schoch S, Quiroz FJU, Rebola N, Bao H, Little JP, Tkachuk AN, 
            Cai E, Hantman AW, Wang SS, DePiero VJ, Borghuis BG, Chapman ER, 
            Dietrich D, DiGregorio DA, Fitzpatrick D, Looger LL
  TITLE     Stability, affinity, and chromatic variants of the glutamate sensor 
            iGluSnFR.
  JOURNAL   Nat Methods. 2018 Nov;15(11):936-939. doi: 
            10.1038/s41592-018-0171-3. Epub 2018 Oct 30.
  PUBMED    30377363
REFERENCE   2  (bases 1 to 6324)
  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"; doi: 
            "10.1038/s41592-018-0171-3"; journalName: "Nat Methods"; date: 
            "2018-11"; volume: "15"; issue: "11"; pages: "936-939"
COMMENT     Sequence Label: pAAV.hSynapsin.SF-iGluSnFR.S72A
FEATURES             Location/Qualifiers
     source          1..6324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   44..173
                     /label=AAV2 ITR
     promoter        249..696
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      725..734
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     731..790
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1759..1780
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1883..1904)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2369..2398
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2402..2548
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2564..3152
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2617..2637)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3035..3046)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3155..3171)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3156..3172)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3201..3322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3212..3231)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3266..3285
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3484..3613
                     /label=AAV2 ITR
     rep_origin      complement(4091..4679)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4171..4190)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4850..5641,5642..5710))
                     /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     5473..5492
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5711..5815)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5841..6296)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5978..5999)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6190..6209
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgcgcg ctcgctcgct
       61 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
      121 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt
      181 aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga tccagggccc
      241 tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg tgcctacctg
      301 acgaccggcc ccgacccact ggacaagcac ccaaccccca ttccccaaat tgcgcatccc
      361 ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact gccagcttca
      421 gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg cctcagcact
      481 gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg ccaccttggt
      541 cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga gatagggggg
      601 cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca gtctgcggtg
      661 ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcca attcaagctg ctagcaagga
      721 tcccgccacc atggagacag acacactcct gctatgggta ctgctgctct gggttccagg
      781 ttccactggt gacagatctg ccgcaggcag cacgctggac aaaatcgcca aaaacggtgt
      841 gattgtcgtc ggtcaccgtg aatcttcagt gcctttctct tattacgaca atcagcaaaa
      901 agtggtgggt tactcgcagg attactccaa cgccattgtt gaagcagtga aaaagaaact
      961 caacaaaccg gacttgcagg taaaactgat tccgattacc gcacaaaacc gtattccact
     1021 gctgcaaaac ggcactttcg attttgaatg tggttctacc accaacaacg tcgaacgcca
     1081 aaaacaggcg gctttctctg acactatttt cgtggtcggt acgcgcctgt tgaccaaaaa
     1141 gggtggcgat atcaaagatt ttgccaacct gaaagacaaa gccgtagtcg tcacttccgg
     1201 cactacctct gaagttttgc tcaacaaact gaatgaagag caaaaaatga atatgcgcat
     1261 catcagcgcc aaagatcacg gtgactcttt ccgcaccctg gaaagcggtc gtgccgttgc
     1321 ctttatgatg gatgacgtgc tgctggccgg tgaacgtgcg aaagcgaaga aaccagacaa
     1381 ctgggaaatc gtcggcaagc cgcagtctca ggaggcctac ggttgtatgt tgcgtaaaga
     1441 tgatccgcag ttcaaaaagc tgatggatga caccatcgct caggtgcaga cctccggtga
     1501 agcggaaaaa tggtttgata agtggttcaa aaatccaatt ctggtgagcc acaacgtcta
     1561 tatcaccgcc gacaagcaga agaacggcat caaggcgaac ttcaagatcc gccacaacgt
     1621 ggaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     1681 ccccgtgctg ctgcccgaca accactacct gagcacccag tccgtgctga gcaaagaccc
     1741 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     1801 cggcatggac gagctgtaca agggcggtac cggagggagc atgagcaagg gcgaggagct
     1861 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     1921 cagcgtgcgc ggcgagggcg agggcgatgc caccaacggc aagctgaccc tgaagttcat
     1981 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     2041 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     2101 catgcccgaa ggctacgtcc aggagcgcac catcagcttc aaggacgacg gcacctacaa
     2161 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     2221 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actttaacaa
     2281 ccccctgaac atgaatttcg aactgtcaga cgaaatgaaa gcactgttca aagaaccgaa
     2341 tgacaaggca ctgaaactgc aggtcgacga acaaaaactc atctcagaag aggatctgaa
     2401 tgctgtgggc caggacacgc aggaggtcat cgtggtgcca cactccttgc cctttaaggt
     2461 ggtggtgatc tcagccatcc tggccctggt ggtgctcacc atcatctccc ttatcatcct
     2521 catcatgctt tggcagaaga agccacgtta gaagcttatc gataatcaac ctctggatta
     2581 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2641 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2701 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2761 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2821 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2881 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2941 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     3001 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3061 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3121 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacctc gacccgggcg
     3181 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3241 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3301 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     3361 gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     3421 taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     3481 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     3541 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     3601 agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3661 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3721 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggrtaacgc
     3781 aggaaagaac atgtgagcaa aargccagca aaargccagg aaccgtaaaa argccgcgkt
     3841 sctggsgtsc ncagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3901 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3961 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4021 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4081 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4141 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4201 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4261 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4321 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4381 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4441 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4501 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4561 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4621 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4681 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4741 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4801 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4861 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4921 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4981 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5041 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5101 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5161 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5221 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5281 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5341 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5401 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5461 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5521 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5581 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5641 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5701 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5761 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5821 ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     5881 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     5941 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     6001 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     6061 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     6121 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     6181 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     6241 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca
     6301 ttcgccattc aggctgcgca actg
//
