LOCUS       sequence-201026-        6845 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Tight affinity glutamate sensor ** A newer version of this sensor is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 6845)
  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 6845)
  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.A184S
FEATURES             Location/Qualifiers
     source          1..6845
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    108..129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(144..149,150..167,168..174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    182..198
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     187..209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     206..222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     206..222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   260..389
                     /label=AAV2 ITR
     promoter        702..979
                     /label=chicken beta-actin promoter
     primer_bind     1259..1281
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1343..1362
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1396..1405
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1402..1461
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     2430..2451
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2554..2575)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3040..3069
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3073..3219
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    3240..3828
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3293..3313)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3711..3722)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3871..3992
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3882..3901)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3936..3955
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4154..4283
                     /label=AAV2 ITR
     primer_bind     complement(4302..4319)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4302..4318)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4311..4333)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4460..4915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4547..4566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4757..4778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4941..5045
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5046..5114,5115..5906)
                     /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(5264..5283)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6077..6665
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6566..6585
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
       61 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      121 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      181 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccagattt
      241 aattaaggcc ttaattaggc tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc
      301 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      361 agtggccaac tccatcacta ggggttcctt gtagttaatg attaacccgc catgctactt
      421 atctacgtag ccatgctcta ggaagatcgt accattgacg tcaataatga cgtatgttcc
      481 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggactatt tacggtaaac
      541 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      601 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      661 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga gccccacgtt
      721 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
      781 ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc caggcggggc
      841 ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag ccaatcagag
      901 cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc cctataaaaa
      961 gcgaagcgcg cggcgggcgg gagtcgctgc gacgctgcct tcgccccgtg ccccgctccg
     1021 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
     1081 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
     1141 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
     1201 gcggctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg
     1261 ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt tcatgccttc
     1321 ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca tcattttggc
     1381 aaagaattgg atcccgccac catggagaca gacacactcc tgctatgggt actgctgctc
     1441 tgggttccag gttccactgg tgacagatct gccgcaggca gcacgctgga caaaatcgcc
     1501 aaaaacggtg tgattgtcgt cggtcaccgt gaatcttcag tgcctttctc ttattacgac
     1561 aatcagcaaa aagtggtggg ttactcgcag gattactcca acgccattgt tgaagcagtg
     1621 aaaaagaaac tcaacaaacc ggacttgcag gtaaaactga ttccgattac ctcacaaaac
     1681 cgtattccac tgctgcaaaa cggcactttc gattttgaat gtggttctac caccaacaac
     1741 gtcgaacgcc aaaaacaggc ggctttctct gacactattt tcgtggtcgg tacgcgcctg
     1801 ttgaccaaaa agggtggcga tatcaaagat tttgccaacc tgaaagacaa agccgtagtc
     1861 gtcacttccg gcactacctc tgaagttttg ctcaacaaac tgaatgaaga gcaaaaaatg
     1921 aatatgcgca tcatcagcgc caaagatcac ggtgactctt tccgcaccct ggaaagcggt
     1981 cgtgccgttg cctttatgat ggatgacagc ctgctggccg gtgaacgtgc gaaagcgaag
     2041 aaaccagaca actgggaaat cgtcggcaag ccgcagtctc aggaggccta cggttgtatg
     2101 ttgcgtaaag atgatccgca gttcaaaaag ctgatggatg acaccatcgc tcaggtgcag
     2161 acctccggtg aagcggaaaa atggtttgat aagtggttca aaaatccaat tctggtgagc
     2221 cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc
     2281 cgccacaacg tggaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
     2341 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgtgctg
     2401 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     2461 gggatcactc tcggcatgga cgagctgtac aagggcggta ccggagggag catgagcaag
     2521 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     2581 ggccacaagt tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg caagctgacc
     2641 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     2701 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     2761 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcagctt caaggacgac
     2821 ggcacctaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     2881 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     2941 aactttaaca accccctgaa catgaatttc gaactgtcag acgaaatgaa agcactgttc
     3001 aaagaaccga atgacaaggc actgaaactg caggtcgacg aacaaaaact catctcagaa
     3061 gaggatctga atgctgtggg ccaggacacg caggaggtca tcgtggtgcc acactccttg
     3121 ccctttaagg tggtggtgat ctcagccatc ctggccctgg tggtgctcac catcatctcc
     3181 cttatcatcc tcatcatgct ttggcagaag aagccacgtt agaagcttag cttatcgata
     3241 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     3301 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     3361 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     3421 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     3481 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     3541 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     3601 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     3661 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     3721 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     3781 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     3841 gacccgggcg gccgcttcga gcagacatga taagatacat tgatgagttt ggacaaacca
     3901 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     3961 ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt
     4021 ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg
     4081 gtaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca tggcgggtta
     4141 atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     4201 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     4261 tcagtgagcg agcgagcgcg cagccttaat taacctaatt cactggccgt cgttttacaa
     4321 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     4381 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     4441 agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4501 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4561 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4621 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4681 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4741 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4801 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4861 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     4921 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4981 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5041 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5101 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5161 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5221 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5281 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5341 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5401 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5461 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5521 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5581 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5641 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5701 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5761 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5821 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5881 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5941 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6001 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6061 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6121 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6181 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6241 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6301 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6361 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6421 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6481 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6541 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6601 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6661 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6721 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6781 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6841 cggaa
//
