LOCUS       sequence-204273-        6931 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DEFINITION  Medium affinity glutamate sensor ** A newer version of this sensor 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 6931)
  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 6931)
  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.A184V
FEATURES             Location/Qualifiers
     source          1..6931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   215..344
                     /label=AAV2 ITR
     intron          937..1069
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1052..1071
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    1082..1115
                     /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    1178..1211
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1239..1385)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1389..1418)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1883..1904
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2007..2028)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2997..3056)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      3053..3062
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3087..3120)
                     /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(3191..3224)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3296..3884
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3349..3369)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3767..3778)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3962..4083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3973..3992)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4027..4046
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4245..4374
                     /label=AAV2 ITR
     rep_origin      complement(4706..5294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4786..4805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5465..6256,6257..6325))
                     /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     6088..6107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6326..6430)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6456..6911)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6593..6614)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6805..6824
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct
       61 cgctcgctca ctgaggccgc ccgggcaaag cccsggssty rscmcmmcwy ycgccgcgct
      121 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg
      181 gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc
      241 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      301 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa
      361 cccgccatgc tacttatcta cgtagccatg ctctaggaag atccgagctc cagtgtgctg
      421 gaattcgccc ttttcagtat ttaaattgac gaccgacccc gacccactgg acaagcaccc
      481 aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg
      541 gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg
      601 cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc
      661 gcaaactccc cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc
      721 gcaccacgcg aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg
      781 cgactcagcg ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc
      841 gcagctgtgc tcctgggcac cgcgcagtcc gcccccgcgg ctcctggcca gaccacccct
      901 aggaccccct gccccaagtc gcagccgttg gatcaggtaa gtatcaaggt tacaagacag
      961 gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga
     1021 taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg tgatgatccc
     1081 cataacttcg tataaagtat cctatacgaa gttatatcaa aataggaaga ccaatgcttc
     1141 accatcgacc cgaattgcca agcatcacca tcgacacata acttcgtata atgtatgcta
     1201 tacgaagtta tactagtgct agcggatccg atatcctaac gtggcttctt ctgccaaagc
     1261 atgatgagga tgataaggga gatgatggtg agcaccacca gggccaggat ggctgagatc
     1321 accaccacct taaagggcaa ggagtgtggc accacgatga cctcctgcgt gtcctggccc
     1381 acagcattca gatcctcttc tgagatgagt ttttgttcgt cgacctgcag tttcagtgcc
     1441 ttgtcattcg gttctttgaa cagtgctttc atttcgtctg acagttcgaa attcatgttc
     1501 agggggttgt taaagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     1561 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     1621 cgggtcttgt aggtgccgtc gtccttgaag ctgatggtgc gctcctggac gtagccttcg
     1681 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac
     1741 tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     1801 gtgcagatga acttcagggt cagcttgccg ttggtggcat cgccctcgcc ctcgccgcgc
     1861 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     1921 gtgaacagct cctcgccctt gctcatgctc cctccggtac cgcccttgta cagctcgtcc
     1981 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
     2041 tcgttggggt ctttgctcag cacggactgg gtgctcaggt agtggttgtc gggcagcagc
     2101 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
     2161 tcctccacgt tgtggcggat cttgaagttc gccttgatgc cgttcttctg cttgtcggcg
     2221 gtgatataga cgttgtggct caccagaatt ggatttttga accacttatc aaaccatttt
     2281 tccgcttcac cggaggtctg cacctgagcg atggtgtcat ccatcagctt tttgaactgc
     2341 ggatcatctt tacgcaacat acaaccgtag gcctcctgag actgcggctt gccgacgatt
     2401 tcccagttgt ctggtttctt cgctttcgca cgttcaccgg ccagcagcac gtcatccatc
     2461 ataaaggcaa cggcacgacc gctttccagg gtgcggaaag agtcaccgtg atctttggcg
     2521 ctgatgatgc gcatattcat tttttgctct tcattcagtt tgttgagcaa aacttcagag
     2581 gtagtgccgg aagtgacgac tacggctttg tctttcaggt tggcaaaatc tttgatatcg
     2641 ccaccctttt tggtcaacag gcgcgtaccg accacgaaaa tagtgtcaga gaaagccgcc
     2701 tgtttttggc gttcgacgtt gttggtggta gaaccacatt caaaatcgaa agtgccgttt
     2761 tgcagcagtg gaatacggtt ttgtgaggta atcggaatca gttttacctg caagtccggt
     2821 ttgttgagtt tctttttcac tgcttcaaca atggcgttgg agtaatcctg cgagtaaccc
     2881 accacttttt gctgattgtc gtaataagag aaaggcactg aagattcacg gtgaccgacg
     2941 acaatcacac cgtttttggc gattttgtcc agcgtgctgc ctgcggcaga tctgtcacca
     3001 gtggaacctg gaacccagag cagcagtacc catagcagga gtgtgtctgt ctccatggtg
     3061 gcgatatcac cggtggtacc actagtataa cttcgtatag gatactttat acgaagttat
     3121 cattgggatt cttcctattt tgatccaagc atcaccatcg accctctagt ccagatctca
     3181 ccatcgaccc ataacttcgt atagcataca ttatacgaag ttatgtccct cgaagaggtt
     3241 cgaaacactg aaaagggcga attctgcaga tgatccactc gagaagctta tcgataatca
     3301 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3361 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3421 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3481 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3541 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3601 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3661 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     3721 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3781 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3841 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     3901 cgggcggccg ctcgagcatg catctagagt cgacccgggc ggccgcttcg agcagacatg
     3961 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4021 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4081 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     4141 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca
     4201 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
     4261 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4321 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat
     4381 taatnsamkc gsmcwrmgss cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4441 gcggcctcag tgagcgagcg agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga
     4501 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4561 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4621 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4681 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4741 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4801 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4861 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4921 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4981 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5041 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5101 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5161 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5221 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5281 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5341 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5401 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5461 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5521 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5581 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5641 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5701 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5761 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5821 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5881 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5941 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6001 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6061 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6121 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6181 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6241 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6301 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6361 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6421 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     6481 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     6541 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     6601 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     6661 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     6721 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     6781 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     6841 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     6901 acagggcgcg tcccattcgc cattcaggct g
//
