LOCUS       sequence-214935-        6905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Medium affinity glutamate sensor ** A newer version of this sensor 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 6905)
  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 6905)
  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.CAG-FLEX.SF-iGluSnFR.A184V
FEATURES             Location/Qualifiers
     source          1..6905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   26..155
                     /label=AAV2 ITR
     promoter        466..743
                     /label=chicken beta-actin promoter
     primer_bind     1022..1044
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1106..1125
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1160..1193
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1256..1289
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1317..1463)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1467..1496)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1961..1982
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2085..2106)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(3075..3134)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      3131..3140
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3165..3198)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(3269..3302)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3374..3962
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3427..3447)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3845..3856)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4005..4126
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4016..4035)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4070..4089
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4288..4417
                     /label=AAV2 ITR
     rep_origin      complement(4654..5242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4734..4753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5413..6204,6205..6273))
                     /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     6036..6055
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6274..6378)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6404..6859)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6541..6562)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6753..6772
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgggcctct tcgctattac gccagctgcg cgctcgctcg ctcactgagg ccgcccgggc
       61 aaagcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
      121 agagggagtg gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg
      181 ctacttatct acgtagccat gctctaggaa gagtaccatt gacgtcaata atgacgtatg
      241 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
      301 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
      361 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
      421 ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag gtgagcccca
      481 cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg tatttattta
      541 ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc gcgccrggsg
      601 gggsggggsg gggsgrgggg sggggsgggg cgaggcggag aggtgcggcg gcagccaatc
      661 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
      721 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
      781 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
      841 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
      901 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
      961 ggagcggctc ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg
     1021 gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca tgttcatgcc
     1081 ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt
     1141 ggcaaagaat tggatcccca taacttcgta taaagtatcc tatacgaagt tatatcaaaa
     1201 taggaagacc aatgcttcac catcgacccg aattgccaag catcaccatc gacccataac
     1261 ttcgtataat gtatgctata cgaagttata ctagtgctag cggatccgat atcctaacgt
     1321 ggcttcttct gccaaagcat gatgaggatg ataagggaga tgatggtgag caccaccagg
     1381 gccaggatgg ctgagatcac caccacctta aagggcaagg agtgtggcac cacgatgacc
     1441 tcctgcgtgt cctggcccac agcattcaga tcctcttctg agatgagttt ttgttcgtcg
     1501 acctgcagtt tcagtgcctt gtcattcggt tctttgaaca gtgctttcat ttcgtctgac
     1561 agttcgaaat tcatgttcag ggggttgtta aagttgtact ccagcttgtg ccccaggatg
     1621 ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc
     1681 tcgaacttca cctcggcgcg ggtcttgtag gtgccgtcgt ccttgaagct gatggtgcgc
     1741 tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg
     1801 gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt gggccagggc
     1861 acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgtt ggtggcatcg
     1921 ccctcgccct cgccgcgcac gctgaacttg tggccgttta cgtcgccgtc cagctcgacc
     1981 aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcatgctccc tccggtaccg
     2041 cccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     2101 accatgtgat cgcgcttctc gttggggtct ttgctcagca cggactgggt gctcaggtag
     2161 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     2221 gcgagctgca cgctgccgtc ctccacgttg tggcggatct tgaagttcgc cttgatgccg
     2281 ttcttctgct tgtcggcggt gatatagacg ttgtggctca ccagaattgg atttttgaac
     2341 cacttatcaa accatttttc cgcttcaccg gaggtctgca cctgagcgat ggtgtcatcc
     2401 atcagctttt tgaactgcgg atcatcttta cgcaacatac aaccgtaggc ctcctgagac
     2461 tgcggcttgc cgacgatttc ccagttgtct ggtttcttcg ctttcgcacg ttcaccggcc
     2521 agcagcacgt catccatcat aaaggcaacg gcacgaccgc tttccagggt gcggaaagag
     2581 tcaccgtgat ctttggcgct gatgatgcgc atattcattt tttgctcttc attcagtttg
     2641 ttgagcaaaa cttcagaggt agtgccggaa gtgacgacta cggctttgtc tttcaggttg
     2701 gcaaaatctt tgatatcgcc accctttttg gtcaacaggc gcgtaccgac cacgaaaata
     2761 gtgtcagaga aagccgcctg tttttggcgt tcgacgttgt tggtggtaga accacattca
     2821 aaatcgaaag tgccgttttg cagcagtgga atacggtttt gtgaggtaat cggaatcagt
     2881 tttacctgca agtccggttt gttgagtttc tttttcactg cttcaacaat ggcgttggag
     2941 taatcctgcg agtaacccac cactttttgc tgattgtcgt aataagagaa aggcactgaa
     3001 gattcacggt gaccgacgac aatcacaccg tttttggcga ttttgtccag cgtgctgcct
     3061 gcggcagatc tgtcaccagt ggaacctgga acccagagca gcagtaccca tagcaggagt
     3121 gtgtctgtct ccatggtggc gatatcaccg gtggtaccac tagtataact tcgtatagga
     3181 tactttatac gaagttatca ttgggattct tcctattttg atccaagcat caccatcgac
     3241 cctctagtcc aaatctcacc atcgacccat aacttcgtat agcatacatt atacgaagtt
     3301 atgtccctcg aagaggttcg aaacactgaa aagggccaat tctgcaaatg atccactcga
     3361 gaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     3421 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     3481 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     3541 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     3601 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     3661 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     3721 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     3781 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     3841 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     3901 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     3961 gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat acattgatga
     4021 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     4081 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     4141 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
     4201 cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac gtagataagt
     4261 agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct
     4321 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     4381 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cattaatgaa tcggccaacg
     4441 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4501 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4561 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4621 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4681 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4741 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4801 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4861 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4921 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4981 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5041 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5101 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5161 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5221 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5281 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5341 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5401 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5461 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5521 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5581 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     5641 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     5701 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5761 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5821 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5881 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5941 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6001 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6061 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6121 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6181 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6241 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6301 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6361 acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat
     6421 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     6481 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     6541 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     6601 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     6661 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     6721 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     6781 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     6841 gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     6901 tcggt
//
