LOCUS       sequence-214726-        6134 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DEFINITION  Tight affinity glutamate sensor ** A newer version of this sensor is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 6134)
  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 6134)
  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.GFAP.SF-iGluSnFR.A184S
FEATURES             Location/Qualifiers
     source          1..6134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      747..756
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     753..812
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1781..1802
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1905..1926)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2391..2420
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2424..2570
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2586..3174
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2639..2659)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3057..3068)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3177..3193)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3178..3194)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3223..3344
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3234..3253)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3288..3307
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3723..4311)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3803..3822)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4482..5273,5274..5342))
                     /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"
     promoter        complement(5343..5447)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5473..5928)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5610..5631)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5822..5841
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6000..6129
                     /label=AAV2 ITR
ORIGIN
        1 ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa gatccaacat
       61 atcctggtgt ggagtagggg acgctgctct gacagaggct cgggggcctg agctggctct
      121 gtgagctggg gaggaggcag acagccaggc ctttgtctgc aagcagacct ggcagcattg
      181 ggctggccgc cccccagggc ctcctcttca tgcccagtga atgactcacc ttggcacaga
      241 cacaatgttc ggggtgggca cagtgcctgc ttcccgccgc accccagccc ccctcaaatg
      301 ccttccgaga agcccattga gcagggggct tgcattgcac cccagcctga cagcctggca
      361 tcttgggata aaagcagcac agccccctag gggctgccct tgctgtgtgg cgccaccggc
      421 ggtggagaac aaggctctat tcagcctgtg cccaggaaag gggatcaggg gatgcccagg
      481 catggacagt gggtggcagg gggggagagg agggctgtct gcttcccaga agtccaagga
      541 cacaaatggg tgaggggaga gctctcccca tagctgggct gcggcccaac cccaccccct
      601 caggctatgc cagggggtgt tgccaggggc acccgggcat cgccagtcta gcccactcct
      661 tcataaagcc ctcgcatccc aggagcgagc agagccagag caggttggag aggagacgca
      721 tcacctccgc tgctcgcggg gatcccgcca ccatggagac agacacactc ctgctatggg
      781 tactgctgct ctgggttcca ggttccactg gtgacagatc tgccgcaggc agcacgctgg
      841 acaaaatcgc caaaaacggt gtgattgtcg tcggtcaccg tgaatcttca gtgcctttct
      901 cttattacga caatcagcaa aaagtggtgg gttactcgca ggattactcc aacgccattg
      961 ttgaagcagt gaaaaagaaa ctcaacaaac cggacttgca ggtaaaactg attccgatta
     1021 cctcacaaaa ccgtattcca ctgctgcaaa acggcacttt cgattttgaa tgtggttcta
     1081 ccaccaacaa cgtcgaacgc caaaaacagg cggctttctc tgacactatt ttcgtggtcg
     1141 gtacgcgcct gttgaccaaa aagggtggcg atatcaaaga ttttgccaac ctgaaagaca
     1201 aagccgtagt cgtcacttcc ggcactacct ctgaagtttt gctcaacaaa ctgaatgaag
     1261 agcaaaaaat gaatatgcgc atcatcagcg ccaaagatca cggtgactct ttccgcaccc
     1321 tggaaagcgg tcgtgccgtt gcctttatga tggatgacag cctgctggcc ggtgaacgtg
     1381 cgaaagcgaa gaaaccagac aactgggaaa tcgtcggcaa gccgcagtct caggaggcct
     1441 acggttgtat gttgcgtaaa gatgatccgc agttcaaaaa gctgatggat gacaccatcg
     1501 ctcaggtgca gacctccggt gaagcggaaa aatggtttga taagtggttc aaaaatccaa
     1561 ttctggtgag ccacaacgtc tatatcaccg ccgacaagca gaagaacggc atcaaggcga
     1621 acttcaagat ccgccacaac gtggaggacg gcagcgtgca gctcgccgac cactaccagc
     1681 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     1741 agtccgtgct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1801 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt accggaggga
     1861 gcatgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1921 gcgacgtaaa cggccacaag ttcagcgtgc gcggcgaggg cgagggcgat gccaccaacg
     1981 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     2041 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     2101 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcagct
     2161 tcaaggacga cggcacctac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     2221 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     2281 agctggagta caactttaac aaccccctga acatgaattt cgaactgtca gacgaaatga
     2341 aagcactgtt caaagaaccg aatgacaagg cactgaaact gcaggtcgac gaacaaaaac
     2401 tcatctcaga agaggatctg aatgctgtgg gccaggacac gcaggaggtc atcgtggtgc
     2461 cacactcctt gccctttaag gtggtggtga tctcagccat cctggccctg gtggtgctca
     2521 ccatcatctc ccttatcatc ctcatcatgc tttggcagaa gaagccacgt tagaagctta
     2581 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     2641 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2701 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2761 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2821 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2881 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2941 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     3001 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     3061 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     3121 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     3181 accgtcgacc tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt
     3241 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3301 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3361 ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc
     3421 tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag
     3481 catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct
     3541 ctgcgcgctc gctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3601 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3661 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3721 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3781 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3841 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3901 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3961 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4021 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4081 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4141 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4201 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4261 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4321 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4381 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4441 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4501 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4561 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4621 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4681 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4741 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4801 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4861 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4921 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4981 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5041 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5101 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5161 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5221 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5281 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5341 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5401 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5461 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     5521 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     5581 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     5641 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     5701 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     5761 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5821 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5881 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
     5941 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     6001 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg
     6061 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
     6121 ggggttcctt gtag
//
