LOCUS       sequence-214898-        6840 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DEFINITION  Medium affinity glutamate sensor ** A newer version of this sensor 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 6840)
  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 6840)
  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.SF-iGluSnFR.A184V
FEATURES             Location/Qualifiers
     source          1..6840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   258..387
                     /label=AAV2 ITR
     promoter        698..975
                     /label=chicken beta-actin promoter
     primer_bind     1254..1276
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1338..1357
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1391..1400
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1397..1456
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     2425..2446
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2549..2570)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3035..3064
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3068..3214
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    3230..3818
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3283..3303)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3701..3712)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3861..3982
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3872..3891)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3926..3945
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4144..4273
                     /label=AAV2 ITR
     rep_origin      4454..4909
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4541..4560)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4751..4772
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4935..5039
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5040..5108,5109..5900)
                     /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     complement(5258..5277)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6071..6659
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6560..6579
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gygmscgagc swkyrcrcmk mrmssgmsss gctyamtscr ccgctacagg
      181 gcgcgtccca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
      241 cttcgctatt acgccagctg cgcgctcgct cgctcactga ggccgcccgg gcaaagcccg
      301 ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc agagagggag
      361 tggccaactc catcactagg ggttccttgt agttaatgat taacccgcca tgctacttat
      421 ctacgtagcc atgctctagg aagagtacca ttgacgtcaa taatgacgta tgttcccata
      481 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      541 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      601 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      661 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
      721 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
      781 ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccrgg sggggsgggg
      841 sggggcgrgg ggsggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc
      901 gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga
      961 agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc
     1021 gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg
     1081 gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt
     1141 ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc
     1201 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
     1261 cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt
     1321 tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga
     1381 attggatccc gccaccatgg agacagacac actcctgcta tgggtactgc tgctctgggt
     1441 tccaggttcc actggtgaca gatctgccgc aggcagcacg ctggacaaaa tcgccaaaaa
     1501 cggtgtgatt gtcgtcggtc accgtgaatc ttcagtgcct ttctcttatt acgacaatca
     1561 gcaaaaagtg gtgggttact cgcaggatta ctccaacgcc attgttgaag cagtgaaaaa
     1621 gaaactcaac aaaccggact tgcaggtaaa actgattccg attacctcac aaaaccgtat
     1681 tccactgctg caaaacggca ctttcgattt tgaatgtggt tctaccacca acaacgtcga
     1741 acgccaaaaa caggcggctt tctctgacac tattttcgtg gtcggtacgc gcctgttgac
     1801 caaaaagggt ggcgatatca aagattttgc caacctgaaa gacaaagccg tagtcgtcac
     1861 ttccggcact acctctgaag ttttgctcaa caaactgaat gaagagcaaa aaatgaatat
     1921 gcgcatcatc agcgccaaag atcacggtga ctctttccgc accctggaaa gcggtcgtgc
     1981 cgttgccttt atgatggatg acgtgctgct ggccggtgaa cgtgcgaaag cgaagaaacc
     2041 agacaactgg gaaatcgtcg gcaagccgca gtctcaggag gcctacggtt gtatgttgcg
     2101 taaagatgat ccgcagttca aaaagctgat ggatgacacc atcgctcagg tgcagacctc
     2161 cggtgaagcg gaaaaatggt ttgataagtg gttcaaaaat ccaattctgg tgagccacaa
     2221 cgtctatatc accgccgaca agcagaagaa cggcatcaag gcgaacttca agatccgcca
     2281 caacgtggag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     2341 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg tgctgagcaa
     2401 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     2461 cactctcggc atggacgagc tgtacaaggg cggtaccgga gggagcatga gcaagggcga
     2521 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2581 caagttcagc gtgcgcggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa
     2641 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2701 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2761 gtccgccatg cccgaaggct acgtccagga gcgcaccatc agcttcaagg acgacggcac
     2821 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2881 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt
     2941 taacaacccc ctgaacatga atttcgaact gtcagacgaa atgaaagcac tgttcaaaga
     3001 accgaatgac aaggcactga aactgcaggt cgacgaacaa aaactcatct cagaagagga
     3061 tctgaatgct gtgggccagg acacgcagga ggtcatcgtg gtgccacact ccttgccctt
     3121 taaggtggtg gtgatctcag ccatcctggc cctggtggtg ctcaccatca tctcccttat
     3181 catcctcatc atgctttggc agaagaagcc acgttagaag cttatcgata atcaacctct
     3241 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3301 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3361 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3421 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3481 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3541 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3601 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     3661 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3721 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3781 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacccgggcg
     3841 gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     3901 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     3961 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4021 gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga
     4081 taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact
     4141 acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     4201 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     4261 agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg gggwsaskgr ggyygggyga
     4321 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     4381 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     4441 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4501 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     4561 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4621 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4681 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4741 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4801 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     4861 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt
     4921 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4981 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5041 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5101 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5161 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5221 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5281 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5341 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5401 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5461 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5521 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5581 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5641 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5701 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5761 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5821 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5881 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5941 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     6001 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     6061 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     6121 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     6181 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6241 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6301 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6361 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6421 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6481 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6541 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6601 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6661 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     6721 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     6781 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
//
