LOCUS       sequence-214869-        6199 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DEFINITION  Medium affinity glutamate sensor ** A newer version of this sensor 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 6199)
  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 6199)
  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.hSynapsin.SF-iGluSnFR.A184V
FEATURES             Location/Qualifiers
     source          1..6199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   55..184
                     /label=AAV2 ITR
     promoter        266..713
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      742..751
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     748..807
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     1776..1797
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1900..1921)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2386..2415
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2419..2565
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    2581..3169
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2634..2654)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3052..3063)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3172..3188)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3173..3189)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3218..3339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3229..3248)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3283..3302
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3501..3630
                     /label=AAV2 ITR
     rep_origin      complement(3844..4432)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3924..3943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4603..5394,5395..5463))
                     /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     5226..5245
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5464..5568)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5594..6049)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5731..5752)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5943..5962
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctgcgc
       61 gctcgctcgc tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc
      121 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
      181 tccttgtagt taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag
      241 atctctgcag agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg
      301 gtgggggtgc ctacctgacg accgaccccg acccactgga caagcaccca acccccattc
      361 cccaaattgc gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt
      421 gcgcactgcc agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc
      481 accgccgcct cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg caaactcccc
      541 ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga
      601 ggcgcgagat aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc
      661 tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagtcgaatt
      721 caagctgcta gcaaggatcc cgccaccatg gagacagaca cactcctgct atgggtactg
      781 ctgctctggg ttccaggttc cactggtgac agatctgccg caggcagcac gctggacaaa
      841 atcgccaaaa acggtgtgat tgtcgtcggt caccgtgaat cttcagtgcc tttctcttat
      901 tacgacaatc agcaaaaagt ggtgggttac tcgcaggatt actccaacgc cattgttgaa
      961 gcagtgaaaa agaaactcaa caaaccggac ttgcaggtaa aactgattcc gattacctca
     1021 caaaaccgta ttccactgct gcaaaacggc actttcgatt ttgaatgtgg ttctaccacc
     1081 aacaacgtcg aacgccaaaa acaggcggct ttctctgaca ctattttcgt ggtcggtacg
     1141 cgcctgttga ccaaaaaggg tggcgatatc aaagattttg ccaacctgaa agacaaagcc
     1201 gtagtcgtca cttccggcac tacctctgaa gttttgctca acaaactgaa tgaagagcaa
     1261 aaaatgaata tgcgcatcat cagcgccaaa gatcacggtg actctttccg caccctggaa
     1321 agcggtcgtg ccgttgcctt tatgatggat gacgtgctgc tggccggtga acgtgcgaaa
     1381 gcgaagaaac cagacaactg ggaaatcgtc ggcaagccgc agtctcagga ggcctacggt
     1441 tgtatgttgc gtaaagatga tccgcagttc aaaaagctga tggatgacac catcgctcag
     1501 gtgcagacct ccggtgaagc ggaaaaatgg tttgataagt ggttcaaaaa tccaattctg
     1561 gtgagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggcgaacttc
     1621 aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1681 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1741 gtgctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1801 gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg agggagcatg
     1861 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1921 gtaaacggcc acaagttcag cgtgcgcggc gagggcgagg gcgatgccac caacggcaag
     1981 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     2041 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     2101 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cagcttcaag
     2161 gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     2221 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     2281 gagtacaact ttaacaaccc cctgaacatg aatttcgaac tgtcagacga aatgaaagca
     2341 ctgttcaaag aaccgaatga caaggcactg aaactgcagg tcgacgaaca aaaactcatc
     2401 tcagaagagg atctgaatgc tgtgggccag gacacgcagg aggtcatcgt ggtgccacac
     2461 tccttgccct ttaaggtggt ggtgatctca gccatcctgg ccctggtggt gctcaccatc
     2521 atctccctta tcatcctcat catgctttgg cagaagaagc cacgttagaa gcttatcgat
     2581 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     2641 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     2701 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     2761 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     2821 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     2881 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     2941 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     3001 gcctgtgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     3061 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     3121 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt
     3181 cgacctcgac ccgggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     3241 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     3301 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     3361 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac
     3421 aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg
     3481 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
     3541 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     3601 ggcggcctca gtgagcgagc gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga
     3661 tcgcccttcc caacagttgc scarcyygmw ksgckmrkkg gwcssgcysy gkmgmgcggt
     3721 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3781 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3841 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3901 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3961 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4021 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4081 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4141 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4201 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4261 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4321 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4381 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4441 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4501 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4561 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4621 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4681 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4741 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4801 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4861 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4921 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4981 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5041 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5101 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5161 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5221 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5281 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5341 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5401 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5461 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5521 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5581 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     5641 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5701 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5761 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5821 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5881 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5941 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     6001 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     6061 ttcaggctgc gcaactgttg gkwmskgcgw tmksysckgr yyyygtsgmt awccgtatta
     6121 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6181 tgagcgagga agcggaaga
//
