LOCUS       sequence-214796-        6908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tight affinity yellow glutamate sensor ** A newer version of this 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 6908)
  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 6908)
  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.hSynap-FLEX.SF-Venus-iGluSnFR.A184S
FEATURES             Location/Qualifiers
     source          1..6908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   215..344
                     /label=AAV2 ITR
     intron          937..1069
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1052..1071
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    1082..1115
                     /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    1178..1211
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1239..1385)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1389..1418)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1883..1904
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2007..2028)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(2997..3056)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      3053..3062
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3087..3120)
                     /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(3191..3224)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3296..3884
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3349..3369)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3767..3778)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3962..4083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3973..3992)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4027..4046
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4683..5271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4763..4782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5442..6233,6234..6302))
                     /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     6065..6084
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6303..6407)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6433..6888)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6570..6591)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6782..6801
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct
       61 cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gsscsmmcwy ysgycgcgct
      121 taatgcgccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact gttgggaagg
      181 gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc tcactgaggc
      241 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
      301 agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa
      361 cccgccatgc tacttatcta cgtagccatg ctctaggaag atccgagctc cagtgtgctg
      421 gaattcgccc ttttcagtat ttaaattgac gaccgacccc gacccactgg acaagcaccc
      481 aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa acaggatgcg
      541 gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc ccccgcctgg
      601 cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg acgtcactcg ccggtccccc
      661 gcaaactccc cttcccggcc accttggtcg cgtccgcgcc gccgccggcc cagccggacc
      721 gcaccacgcg aggcgcgaga taggggggca cgggcgcgac catctgcgct gcggcgccgg
      781 cgactcagcg ctgcctcagt ctgcggtggg cagcggagga gtcgtgtcgt gcctgagagc
      841 gcagctgtgc tcctgggcac cgcgcagtcc gcccccgcgg ctcctggcca gaccacccct
      901 aggaccccct gccccaagtc gcagccgttg gatcaggtaa gtatcaaggt tacaagacag
      961 gtttaaggag accaatagaa actgggcttg tcgagacaga gaagactctt gcgtttctga
     1021 taggcaccta ttggtcttac tgacatccac tttgcctttc tctccacagg tgatgatccc
     1081 cataacttcg tataaagtat cctatacgaa gttatatcaa aataggaaga ccaatgcttc
     1141 accatcgacc cgaattgcca agcatcacca tcgacacata acttcgtata atgtatgcta
     1201 tacgaagtta tactagtgct agcggatccg atatcctaac gtggcttctt ctgccaaagc
     1261 atgatgagga tgataaggga gatgatggtg agcaccacca gggccaggat ggctgagatc
     1321 accaccacct taaagggcaa ggagtgtggc accacgatga cctcctgcgt gtcctggccc
     1381 acagcattca gatcctcttc tgagatgagt ttttgttcgt cgacctgcag tttcagtgcc
     1441 ttgtcattcg gttctttgaa cagtgctttc atttcgtctg acagttcgaa attcatgttc
     1501 agggggttgt taaagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     1561 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     1621 cgggtcttgt aggtgccgtc gtccttgaag ctgatggtgc gctcctggac gtagccttcg
     1681 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac
     1741 tgcacgccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     1801 gtgcagatga gcttcagggt cagcttgccg ttggtggcat cgccctcgcc ctcgccgcgc
     1861 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     1921 gtgaacagct cctcgccctt gctcatgctc cctccggtac cgcccttgta cagctcgtcc
     1981 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
     2041 tcgttggggt ctttgctcag cacggactga tagctcaggt agtggttgtc gggcagcagc
     2101 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
     2161 tcctccacgt tgtggcggat cttgaagttc gccttgatgc cgttcttctg cttgtcggcg
     2221 gtgatataga cgttgtggct caccagaatt ggatttttga accacttatc aaaccatttt
     2281 tccgcttcac cggaggtctg cacctgagcg atggtgtcat ccatcagctt tttgaactgc
     2341 ggatcatctt tacgcaacat acaaccgtag gcctcctgag actgcggctt gccgacgatt
     2401 tcccagttgt ctggtttctt cgctttcgca cgttcaccgg ccagcaggct gtcatccatc
     2461 ataaaggcaa cggcacgacc gctttccagg gtgcggaaag agtcaccgtg atctttggcg
     2521 ctgatgatgc gcatattcat tttttgctct tcattcagtt tgttgagcaa aacttcagag
     2581 gtagtgccgg aagtgacgac tacggctttg tctttcaggt tggcaaaatc tttgatatcg
     2641 ccaccctttt tggtcaacag gcgcgtaccg accacgaaaa tagtgtcaga gaaagccgcc
     2701 tgtttttggc gttcgacgtt gttggtggta gaaccacatt caaaatcgaa agtgccgttt
     2761 tgcagcagtg gaatacggtt ttgtgaggta atcggaatca gttttacctg caagtccggt
     2821 ttgttgagtt tctttttcac tgcttcaaca atggcgttgg agtaatcctg cgagtaaccc
     2881 accacttttt gctgattgtc gtaataagag aaaggcactg aagattcacg gtgaccgacg
     2941 acaatcacac cgtttttggc gattttgtcc agcgtgctgc ctgcggcaga tctgtcacca
     3001 gtggaacctg gaacccagag cagcagtacc catagcagga gtgtgtctgt ctccatggtg
     3061 gcgatatcac cggtggtacc actagtataa cttcgtatag gatactttat acgaagttat
     3121 cattgggatt cttcctattt tgatccaagc atcaccatcg accctctagt ccaaatctca
     3181 ccatcgaccc ataacttcgt atagcataca ttatacgaag ttatgtccct cgaagaggtt
     3241 cgaaacactg aaaagggcca attctgcaaa tgatccactc gagaagctta tcgataatca
     3301 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3361 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3421 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     3481 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     3541 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     3601 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     3661 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg
     3721 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     3781 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     3841 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgtcgacc
     3901 cgggcggccg ctcgagcatg catctagagt cgacccgggc ggccgcttcg agcagacatg
     3961 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4021 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4081 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     4141 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca
     4201 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
     4261 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggacgccc gggctttgcc
     4321 cgggcggcct cagtgascgr gmrmscsyrs wgmtgsagtw rksmaytcsc ycwctgcgcg
     4381 skcgctcgct cactgaggac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4441 gcgcagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4501 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4561 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4621 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4681 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4741 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4801 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4861 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4921 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4981 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5041 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5101 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5161 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5221 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5281 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5341 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5401 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5461 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5521 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5581 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5641 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5701 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5761 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5821 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5881 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5941 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     6001 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     6061 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6121 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6181 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6241 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6301 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6361 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6421 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     6481 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     6541 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     6601 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     6661 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     6721 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     6781 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     6841 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
     6901 tcaggctg
//
