LOCUS       sequence-204613-        7468 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DEFINITION  Tight affinity glutamate sensor ** A newer version of this sensor is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 7468)
  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 7468)
  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.A184S.mRuby3
FEATURES             Location/Qualifiers
     source          1..7468
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     417..439
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     501..520
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    555..588
                     /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    651..684
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1447..1593)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1597..1626)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     2091..2112
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2215..2236)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(3205..3264)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      3261..3270
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3295..3328)
                     /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(3399..3432)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3504..4092
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3557..3577)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3975..3986)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4135..4256
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4146..4165)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4200..4219
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4635..5223)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4715..4734)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5394..6185,6186..6254))
                     /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     6017..6036
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6255..6359)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6385..6840)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6522..6543)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6734..6753
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6912..7041
                     /label=AAV2 ITR
     promoter        join(7352..7468,1..138)
                     /label=chicken beta-actin promoter
                     /note="contains a 21-bp deletion relative to the standard 
                     chicken beta-actin promoter"
ORIGIN
        1 gggcggggcg aggggcgggg cggggcgagg cggagaggtg cggcggcagc caatcagagc
       61 ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag
      121 cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc
      181 gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg
      241 cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct
      301 tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc
      361 ggctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt
      421 cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt
      481 ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa
      541 agaattggat ccccataact tcgtataaag tatcctatac gaagttatat caaaatagga
      601 agaccaatgc ttcaccatcg acccgaattg ccaagcatca ccatcgaccc ataacttcgt
      661 ataatgtatg ctatacgaag ttatactagt gctagcggat ccgatatcct aaccaccaag
      721 attggagtac ttggcaactg cgacttcgcg ttgtactacg tacgtttcat tgtcactttc
      781 ctctatcctt tccaggcggt gatcaacggc atggacaccg ggcatcttga tgttcccgac
      841 ggtctttttt gacctgtaag ttgttacgaa attgcaatgc agatggccac caccatcaac
      901 tttcagtgca atatcagtgt atcccctcag accaccatct gctggataca tcatctctgt
      961 attaggctcc caacccttgg tcttcttctg catcacggga ccattggagg gaaagtttac
     1021 ccctctgact ttgacgttgt aaacgagttc gccatcctca aggctggtgt cctgcgtgac
     1081 ggtgacgact ccaccatctt cgtatctcgt aactctttcc caagtaaaac cctcaggaaa
     1141 ggactgttta aagaaatcag gaatgtctgc cgggtacttg ataaaagtac ggctgccata
     1201 catgaacgac gtggcaagaa tgtcaaaggc aaatggcagg ggtcctccct cgatgacttt
     1261 gatcctcatg gtttgcactc cctcgtacgg cctgccttct ccttcacctg tgcatttgaa
     1321 ttggtggccg ttgaccgaac cttccatgac caccttcata cgcatatttt ccttgatcag
     1381 ctcttcgccc ttagacacca tacgctcctg cgtgtcctgg cccacagcat ttccggagcc
     1441 tctagaacgt ggcttcttct gccaaagcat gatgaggatg ataagggaga tgatggtgag
     1501 caccaccagg gccaggatgg ctgagatcac caccacctta aagggcaagg agtgtggcac
     1561 cacgatgacc tcctgcgtgt cctggcccac agcattcaga tcctcttctg agatgagttt
     1621 ttgttcgtcg acctgcagtt tcagtgcctt gtcattcggt tctttgaaca gtgctttcat
     1681 ttcgtctgac agttcgaaat tcatgttcag ggggttgtta aagttgtact ccagcttgtg
     1741 ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc agctcgatgc ggttcaccag
     1801 ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag gtgccgtcgt ccttgaagct
     1861 gatggtgcgc tcctggacgt agccttcggg catggcggac ttgaagaagt cgtgctgctt
     1921 catgtggtcg gggtagcggc tgaagcactg cacgccgtag gtcagggtgg tcacgagggt
     1981 gggccagggc acgggcagct tgccggtggt gcagatgaac ttcagggtca gcttgccgtt
     2041 ggtggcatcg ccctcgccct cgccgcgcac gctgaacttg tggccgttta cgtcgccgtc
     2101 cagctcgacc aggatgggca ccaccccggt gaacagctcc tcgcccttgc tcatgctccc
     2161 tccggtaccg cccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa
     2221 ctccagcagg accatgtgat cgcgcttctc gttggggtct ttgctcagca cggactgggt
     2281 gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg
     2341 gtagtggtcg gcgagctgca cgctgccgtc ctccacgttg tggcggatct tgaagttcgc
     2401 cttgatgccg ttcttctgct tgtcggcggt gatatagacg ttgtggctca ccagaattgg
     2461 atttttgaac cacttatcaa accatttttc cgcttcaccg gaggtctgca cctgagcgat
     2521 ggtgtcatcc atcagctttt tgaactgcgg atcatcttta cgcaacatac aaccgtaggc
     2581 ctcctgagac tgcggcttgc cgacgatttc ccagttgtct ggtttcttcg ctttcgcacg
     2641 ttcaccggcc agcaggctgt catccatcat aaaggcaacg gcacgaccgc tttccagggt
     2701 gcggaaagag tcaccgtgat ctttggcgct gatgatgcgc atattcattt tttgctcttc
     2761 attcagtttg ttgagcaaaa cttcagaggt agtgccggaa gtgacgacta cggctttgtc
     2821 tttcaggttg gcaaaatctt tgatatcgcc accctttttg gtcaacaggc gcgtaccgac
     2881 cacgaaaata gtgtcagaga aagccgcctg tttttggcgt tcgacgttgt tggtggtaga
     2941 accacattca aaatcgaaag tgccgttttg cagcagtgga atacggtttt gtgaggtaat
     3001 cggaatcagt tttacctgca agtccggttt gttgagtttc tttttcactg cttcaacaat
     3061 ggcgttggag taatcctgcg agtaacccac cactttttgc tgattgtcgt aataagagaa
     3121 aggcactgaa gattcacggt gaccgacgac aatcacaccg tttttggcga ttttgtccag
     3181 cgtgctgcct gcggcagatc tgtcaccagt ggaacctgga acccagagca gcagtaccca
     3241 tagcaggagt gtgtctgtct ccatggtggc gatatcaccg gtggtaccac tagtataact
     3301 tcgtatagga tactttatac gaagttatca ttgggattct tcctattttg atccaagcat
     3361 caccatcgac cctctagtcc aaatctcacc atcgacccat aacttcgtat agcatacatt
     3421 atacgaagtt atgtccctcg aagaggttcg aaacactgaa aagggccaat tctgcaaatg
     3481 atccactcga gaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     3541 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     3601 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3661 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3721 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3781 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3841 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     3901 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     3961 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     4021 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     4081 ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat
     4141 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     4201 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     4261 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     4321 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac
     4381 gtagataagt agcatggcgg gttaatcatt aactacaagg aacccctagt gatggagttg
     4441 gccactccct ctctgcgcgc tcgctcgctc actgactcgc tgcgctcggt cgttcggctg
     4501 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4561 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4621 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4681 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4741 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4801 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4861 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4921 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4981 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     5041 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     5101 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     5161 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     5221 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5281 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5341 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5401 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5461 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5521 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5581 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5641 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5701 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5761 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5821 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5881 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5941 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     6001 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     6061 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     6121 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     6181 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6241 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6301 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6361 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     6421 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     6481 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6541 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     6601 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     6661 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     6721 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     6781 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     6841 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     6901 tattacgcca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
     6961 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     7021 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt
     7081 agccatgctc taggaagagt accattgacg tcaataatga cgtatgttcc catagtaacg
     7141 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     7201 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     7261 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     7321 atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact
     7381 ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt
     7441 tgtgcagcga tgggggcggg gggggggg
//
