LOCUS       sequence-214721-        6134 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DEFINITION  Medium affinity glutamate sensor ** A newer version of this sensor 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 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.A184V
FEATURES             Location/Qualifiers
     source          1..6134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      841..850
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     847..906
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1875..1896
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1999..2020)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2485..2514
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2518..2664
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2680..3268
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2733..2753)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3151..3162)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3271..3287)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3272..3288)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3317..3438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3328..3347)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3382..3401
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(3817..4405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3897..3916)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4576..5367,5368..5436))
                     /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(5437..5541)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5567..6022)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5704..5725)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5916..5935
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(6094..6134,1..89)
                     /label=AAV2 ITR
ORIGIN
        1 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
       61 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      121 atctacgtag ccatgctcta ggaagatcca acatatcctg gtgtggagta ggggacgctg
      181 ctctgacaga ggctcggggg cctgagctgg ctctgtgagc tggggaggag gcagacagcc
      241 aggcctttgt ctgcaagcag acctggcagc attgggctgg ccgcccccca gggcctcctc
      301 ttcatgccca gtgaatgact caccttggca cagacacaat gttcggggtg ggcacagtgc
      361 ctgcttcccg ccgcacccca gcccccctca aatgccttcc gagaagccca ttgagcaggg
      421 ggcttgcatt gcaccccagc ctgacagcct ggcatcttgg gataaaagca gcacagcccc
      481 ctaggggctg cccttgctgt gtggcgccac cggcggtgga gaacaaggct ctattcagcc
      541 tgtgcccagg aaaggggatc aggggatgcc caggcatgga cagtgggtgg caggggggga
      601 gaggagggct gtctgcttcc cagaagtcca aggacacaaa tgggtgaggg gagagctctc
      661 cccatagctg ggctgcggcc caaccccacc ccctcaggct atgccagggg gtgttgccag
      721 gggcacccgg gcatcgccag tctagcccac tccttcataa agccctcgca tcccaggagc
      781 gagcagagcc agagcaggtt ggagaggaga cgcatcacct ccgctgctcg cggggatccc
      841 gccaccatgg agacagacac actcctgcta tgggtactgc tgctctgggt tccaggttcc
      901 actggtgaca gatctgccgc aggcagcacg ctggacaaaa tcgccaaaaa cggtgtgatt
      961 gtcgtcggtc accgtgaatc ttcagtgcct ttctcttatt acgacaatca gcaaaaagtg
     1021 gtgggttact cgcaggatta ctccaacgcc attgttgaag cagtgaaaaa gaaactcaac
     1081 aaaccggact tgcaggtaaa actgattccg attacctcac aaaaccgtat tccactgctg
     1141 caaaacggca ctttcgattt tgaatgtggt tctaccacca acaacgtcga acgccaaaaa
     1201 caggcggctt tctctgacac tattttcgtg gtcggtacgc gcctgttgac caaaaagggt
     1261 ggcgatatca aagattttgc caacctgaaa gacaaagccg tagtcgtcac ttccggcact
     1321 acctctgaag ttttgctcaa caaactgaat gaagagcaaa aaatgaatat gcgcatcatc
     1381 agcgccaaag atcacggtga ctctttccgc accctggaaa gcggtcgtgc cgttgccttt
     1441 atgatggatg acgtgctgct ggccggtgaa cgtgcgaaag cgaagaaacc agacaactgg
     1501 gaaatcgtcg gcaagccgca gtctcaggag gcctacggtt gtatgttgcg taaagatgat
     1561 ccgcagttca aaaagctgat ggatgacacc atcgctcagg tgcagacctc cggtgaagcg
     1621 gaaaaatggt ttgataagtg gttcaaaaat ccaattctgg tgagccacaa cgtctatatc
     1681 accgccgaca agcagaagaa cggcatcaag gcgaacttca agatccgcca caacgtggag
     1741 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1801 gtgctgctgc ccgacaacca ctacctgagc acccagtccg tgctgagcaa agaccccaac
     1861 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1921 atggacgagc tgtacaaggg cggtaccgga gggagcatga gcaagggcga ggagctgttc
     1981 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2041 gtgcgcggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc
     2101 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
     2161 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2221 cccgaaggct acgtccagga gcgcaccatc agcttcaagg acgacggcac ctacaagacc
     2281 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2341 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt taacaacccc
     2401 ctgaacatga atttcgaact gtcagacgaa atgaaagcac tgttcaaaga accgaatgac
     2461 aaggcactga aactgcaggt cgacgaacaa aaactcatct cagaagagga tctgaatgct
     2521 gtgggccagg acacgcagga ggtcatcgtg gtgccacact ccttgccctt taaggtggtg
     2581 gtgatctcag ccatcctggc cctggtggtg ctcaccatca tctcccttat catcctcatc
     2641 atgctttggc agaagaagcc acgttagaag cttatcgata atcaacctct ggattacaaa
     2701 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     2761 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     2821 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     2881 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     2941 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3001 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3061 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     3121 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3181 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3241 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgacc cgggcggccg
     3301 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     3361 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     3421 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     3481 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     3541 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     3601 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgactc
     3661 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3721 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3781 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3841 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3901 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3961 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4021 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4081 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4141 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4201 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4261 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4321 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4381 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4441 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4501 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4561 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4621 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4681 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4741 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4801 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4861 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4921 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4981 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5041 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5101 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5161 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5221 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5281 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5341 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5401 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5461 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5521 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     5581 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     5641 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     5701 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     5761 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     5821 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     5881 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     5941 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     6001 aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg
     6061 cgatcggtgc gggcctcttc gctattacgc cagctgcgcg ctcgctcgct cactgaggcc
     6121 gcccgggcaa agcc
//
