LOCUS       sequence-246155-        6651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tight affinity glutamate sensor ** A newer version of this sensor is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 6651)
  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 6651)
  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.hSynap-FLEX.SF-iGluSnFR.A184S
FEATURES             Location/Qualifiers
     source          1..6651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   52..181
                     /label=AAV2 ITR
     intron          774..906
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     889..908
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    919..952
                     /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    1015..1048
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1076..1222)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1226..1255)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1720..1741
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1844..1865)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2834..2893)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2890..2899
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2924..2957)
                     /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(3028..3061)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3133..3721
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3186..3206)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3604..3615)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3799..3920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3810..3829)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3864..3883
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4426..5014)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4506..4525)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5185..5976,5977..6045))
                     /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     5808..5827
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6046..6150)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6176..6631)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6313..6334)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6525..6544
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct
       61 cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg
      121 gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc
      181 ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc taggaagatc
      241 cgagctccag tgtgctggaa ttcgcccttt tcagtattta aattgacgac cgaccccgac
      301 ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg
      361 aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg
      421 ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg
      481 tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc
      541 gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat
      601 ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc
      661 gtgtcgtgcc tgagagcgca gctgtgctcc tgggcaccgc gcagtccgcc cccgcggctc
      721 ctggccagac cacccctagg accccctgcc ccaagtcgca gccgttggat caggtaagta
      781 tcaaggttac aagacaggtt taaggagacc aatagaaact gggcttgtcg agacagagaa
      841 gactcttgcg tttctgatag gcacctattg gtcttactga catccacttt gcctttctct
      901 ccacaggtga tgatccccat aacttcgtat aaagtatcct atacgaagtt atatcaaaat
      961 aggaagacca atgcttcacc atcgacccga attgccaagc atcaccatcg acacataact
     1021 tcgtataatg tatgctatac gaagttatac tagtgctagc ggatccgata tcctaacgtg
     1081 gcttcttctg ccaaagcatg atgaggatga taagggagat gatggtgagc accaccaggg
     1141 ccaggatggc tgagatcacc accaccttaa agggcaagga gtgtggcacc acgatgacct
     1201 cctgcgtgtc ctggcccaca gcattcagat cctcttctga gatgagtttt tgttcgtcga
     1261 cctgcagttt cagtgccttg tcattcggtt ctttgaacag tgctttcatt tcgtctgaca
     1321 gttcgaaatt catgttcagg gggttgttaa agttgtactc cagcttgtgc cccaggatgt
     1381 tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct
     1441 cgaacttcac ctcggcgcgg gtcttgtagg tgccgtcgtc cttgaagctg atggtgcgct
     1501 cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg
     1561 ggtagcggct gaagcactgc acgccgtagg tcagggtggt cacgagggtg ggccagggca
     1621 cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgttg gtggcatcgc
     1681 cctcgccctc gccgcgcacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca
     1741 ggatgggcac caccccggtg aacagctcct cgcccttgct catgctccct ccggtaccgc
     1801 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     1861 ccatgtgatc gcgcttctcg ttggggtctt tgctcagcac ggactgggtg ctcaggtagt
     1921 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
     1981 cgagctgcac gctgccgtcc tccacgttgt ggcggatctt gaagttcgcc ttgatgccgt
     2041 tcttctgctt gtcggcggtg atatagacgt tgtggctcac cagaattgga tttttgaacc
     2101 acttatcaaa ccatttttcc gcttcaccgg aggtctgcac ctgagcgatg gtgtcatcca
     2161 tcagcttttt gaactgcgga tcatctttac gcaacataca accgtaggcc tcctgagact
     2221 gcggcttgcc gacgatttcc cagttgtctg gtttcttcgc tttcgcacgt tcaccggcca
     2281 gcaggctgtc atccatcata aaggcaacgg cacgaccgct ttccagggtg cggaaagagt
     2341 caccgtgatc tttggcgctg atgatgcgca tattcatttt ttgctcttca ttcagtttgt
     2401 tgagcaaaac ttcagaggta gtgccggaag tgacgactac ggctttgtct ttcaggttgg
     2461 caaaatcttt gatatcgcca ccctttttgg tcaacaggcg cgtaccgacc acgaaaatag
     2521 tgtcagagaa agccgcctgt ttttggcgtt cgacgttgtt ggtggtagaa ccacattcaa
     2581 aatcgaaagt gccgttttgc agcagtggaa tacggttttg tgaggtaatc ggaatcagtt
     2641 ttacctgcaa gtccggtttg ttgagtttct ttttcactgc ttcaacaatg gcgttggagt
     2701 aatcctgcga gtaacccacc actttttgct gattgtcgta ataagagaaa ggcactgaag
     2761 attcacggtg accgacgaca atcacaccgt ttttggcgat tttgtccagc gtgctgcctg
     2821 cggcagatct gtcaccagtg gaacctggaa cccagagcag cagtacccat agcaggagtg
     2881 tgtctgtctc catggtggcg atatcaccgg tggtaccact agtataactt cgtataggat
     2941 actttatacg aagttatcat tgggattctt cctattttga tccaagcatc accatcgacc
     3001 ctctagtcca aatctcacca tcgacccata acttcgtata gcatacatta tacgaagtta
     3061 tgtccctcga agaggttcga aacactgaaa agggccaatt ctgcaaatga tccactcgag
     3121 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     3181 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     3241 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     3301 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     3361 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     3421 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     3481 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     3541 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     3601 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     3661 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     3721 catcgatacc gtcgacccgg gcggccgctc gagcatgcat ctagagtcga cccgggcggc
     3781 cgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     3841 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     3901 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     3961 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata
     4021 aggatcttcc tagagcatgg ctacgtagat aagtagcatg gcgggttaat cattaactac
     4081 aaggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     4141 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg
     4201 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     4261 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     4321 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4381 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4441 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4501 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4561 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4621 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4681 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4741 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4801 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4861 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4921 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4981 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5041 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5101 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5161 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5221 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5281 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5341 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5401 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5461 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5521 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5581 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5641 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5701 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5761 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5821 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5881 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5941 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     6001 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6061 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6121 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt
     6181 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     6241 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     6301 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     6361 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     6421 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     6481 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     6541 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     6601 gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct g
//
