LOCUS       sequence-204216-        6905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Medium affinity yellow glutamate sensor ** A newer version of this 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 6905)
  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 6905)
  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-FLEX.SF-Venus-iGluSnFR.A184V
FEATURES             Location/Qualifiers
     source          1..6905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   17..146
                     /label=AAV2 ITR
     promoter        457..734
                     /label=chicken beta-actin promoter
     primer_bind     1013..1035
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1097..1116
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1151..1184
                     /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    1247..1280
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(1308..1454)
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             complement(1458..1487)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     1952..1973
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2076..2097)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     sig_peptide     complement(3066..3125)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      3122..3131
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(3156..3189)
                     /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(3260..3293)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3365..3953
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3418..3438)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3836..3847)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3996..4117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4007..4026)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4061..4080
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4279..4408
                     /label=AAV2 ITR
     rep_origin      complement(4645..5233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4725..4744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5404..6195,6196..6264))
                     /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     6027..6046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6265..6369)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6395..6850)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6532..6553)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6744..6763
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttcgctatta cgccagctgc gcgctcgctc gctcactgag gccgcccggg caaagcccgg
       61 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
      121 ggccaactcc atcactaggg gttccttgta gttaatgatt aacccgccat gctacttatc
      181 tacgtagcca tgctctagga agagtaccat tgacgtcaat aatgacgtat gttcccatag
      241 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      301 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      361 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      421 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
      481 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
      541 tattttgtgc agcgatgggg gcgggggggg ggggggggcg cgcgccaggc ggggcggggc
      601 ggggcgrggg gsggggcggg gcgaggcgga gaggtgcggc ggcagccaat cagagcggcg
      661 cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg gcggccctat aaaaagcgaa
      721 gcgcgcggcg ggcgggagtc gctgcgcgct gccttcgccc cgtgccccgc tccgccgccg
      781 cctcgcgccg cccgccccgg ctctgactga ccgcgttact cccacaggtg agcgggcggg
      841 acggcccttc tcctccgggc tgtaattagc gcttggttta atgacggctt gtttcttttc
      901 tgtggctgcg tgaaagcctt gaggggctcc gggagggccc tttgtgcggg gggagcggct
      961 cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc ggggttcggc
     1021 ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc cttcttcttt
     1081 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa
     1141 ttggatcccc ataacttcgt ataaagtatc ctatacgaag ttatatcaaa ataggaagac
     1201 caatgcttca ccatcgaccc gaattgccaa gcatcaccat cgacccataa cttcgtataa
     1261 tgtatgctat acgaagttat actagtgcta gcggatccga tatcctaacg tggcttcttc
     1321 tgccaaagca tgatgaggat gataagggag atgatggtga gcaccaccag ggccaggatg
     1381 gctgagatca ccaccacctt aaagggcaag gagtgtggca ccacgatgac ctcctgcgtg
     1441 tcctggccca cagcattcag atcctcttct gagatgagtt tttgttcgtc gacctgcagt
     1501 ttcagtgcct tgtcattcgg ttctttgaac agtgctttca tttcgtctga cagttcgaaa
     1561 ttcatgttca gggggttgtt aaagttgtac tccagcttgt gccccaggat gttgccgtcc
     1621 tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc
     1681 acctcggcgc gggtcttgta ggtgccgtcg tccttgaagc tgatggtgcg ctcctggacg
     1741 tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg
     1801 gcgaagcact gcacgccgta gcccagggtg gtcacgaggg tgggccaggg cacgggcagc
     1861 ttgccggtgg tgcagatgag cttcagggtc agcttgccgt tggtggcatc gccctcgccc
     1921 tcgccgcgca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc
     1981 accaccccgg tgaacagctc ctcgcccttg ctcatgctcc ctccggtacc gcccttgtac
     2041 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
     2101 tcgcgcttct cgttggggtc tttgctcagc acggactgat agctcaggta gtggttgtcg
     2161 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggtc ggcgagctgc
     2221 acgctgccgt cctccacgtt gtggcggatc ttgaagttcg ccttgatgcc gttcttctgc
     2281 ttgtcggcgg tgatatagac gttgtggctc accagaattg gatttttgaa ccacttatca
     2341 aaccattttt ccgcttcacc ggaggtctgc acctgagcga tggtgtcatc catcagcttt
     2401 ttgaactgcg gatcatcttt acgcaacata caaccgtagg cctcctgaga ctgcggcttg
     2461 ccgacgattt cccagttgtc tggtttcttc gctttcgcac gttcaccggc cagcagcacg
     2521 tcatccatca taaaggcaac ggcacgaccg ctttccaggg tgcggaaaga gtcaccgtga
     2581 tctttggcgc tgatgatgcg catattcatt ttttgctctt cattcagttt gttgagcaaa
     2641 acttcagagg tagtgccgga agtgacgact acggctttgt ctttcaggtt ggcaaaatct
     2701 ttgatatcgc cacccttttt ggtcaacagg cgcgtaccga ccacgaaaat agtgtcagag
     2761 aaagccgcct gtttttggcg ttcgacgttg ttggtggtag aaccacattc aaaatcgaaa
     2821 gtgccgtttt gcagcagtgg aatacggttt tgtgaggtaa tcggaatcag ttttacctgc
     2881 aagtccggtt tgttgagttt ctttttcact gcttcaacaa tggcgttgga gtaatcctgc
     2941 gagtaaccca ccactttttg ctgattgtcg taataagaga aaggcactga agattcacgg
     3001 tgaccgacga caatcacacc gtttttggcg attttgtcca gcgtgctgcc tgcggcagat
     3061 ctgtcaccag tggaacctgg aacccagagc agcagtaccc atagcaggag tgtgtctgtc
     3121 tccatggtgg cgatatcacc ggtggtacca ctagtataac ttcgtatagg atactttata
     3181 cgaagttatc attgggattc ttcctatttt gatccaagca tcaccatcga ccctctagtc
     3241 caaatctcac catcgaccca taacttcgta tagcatacat tatacgaagt tatgtccctc
     3301 gaagaggttc gaaacactga aaagggccaa ttctgcaaat gatccactcg agaagcttat
     3361 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3421 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3481 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3541 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3601 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3661 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3721 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     3781 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3841 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3901 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     3961 ccgtcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     4021 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     4081 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4141 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     4201 atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag tagcatggcg
     4261 ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4321 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4381 cggcctcagt gagcgagcga gcgcgcagct gcattaatga atcggccaac gcgcggggag
     4441 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4501 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4561 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4621 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4681 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4741 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4801 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4861 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4921 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4981 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5041 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5101 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5161 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5221 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5281 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5341 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5401 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5461 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5521 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5581 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5641 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5701 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5761 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5821 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5881 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5941 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6001 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6061 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6121 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6181 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6241 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6301 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6361 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     6421 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     6481 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     6541 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     6601 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     6661 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     6721 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     6781 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     6841 cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     6901 gcctc
//
