LOCUS       sequence-487018-        7201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasma membrane targeted expression of green glucose sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7201)
  AUTHORS   Marvin JS, Machler P, Meng C, Ates T, Patel RH, Adhikari R, Makurath
            MA, Ku Z, Feliciano D, Atasoy D, Cui G, Kleinfeld D, Brown TA
  TITLE     iGlucoSnFR2: A genetically encoded fluorescent sensor for measuring 
            intracellular or extracellular glucose in vivo in mouse brain.
  JOURNAL   Sci Adv. 2025 Nov 14;11(46):eadz3889. doi: 10.1126/sciadv.adz3889. 
            Epub 2025 Nov 12.
  PUBMED    41223288
REFERENCE   2  (bases 1 to 7201)
  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"; journalName: "Sci Adv."; 
            date: "2025-11-14"; volume: "11(46):eadz3889. doi"; pages: " 
            10.1126/sciadv.adz3889. Epub 2025 Nov 12"
COMMENT     Sequence Label: pAAV.CAGFLEX.(mem).iGlucoSnFR2
FEATURES             Location/Qualifiers
     source          1..7201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1133..1166
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1237..1270
                     /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)."
     regulatory      1278..1287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1284..1343
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     2540..2561
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2664..2685)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3282..3311
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3315..3461
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(3477..3510)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3573..3606)
                     /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)."
     primer_bind     complement(3644..3663)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3725..3747)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(4026..4303)
                     /label=chicken beta-actin promoter
     repeat_region   4614..4743
                     /label=AAV2 ITR
     rep_origin      4815..5270
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4902..4921)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5112..5133
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5296..5400
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5401..5469,5470..6261)
                     /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(5619..5638)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6432..7020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6921..6940
                     /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 gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct aattcgaacc tcttcgaggg acataacttc
     1141 gtataatgta tgctatacga agttatgggt cgatggtgag atctggacta gagggtcgat
     1201 ggtgatgctt ggatcaaaat aggaagaatc ccaatgataa cttcgtataa agtatcctat
     1261 acgaagttat ggatcccgcc accatggaga cagacacact cctgctatgg gtactgctgc
     1321 tctgggttcc aggttccact ggtgacagat ctaagctgga gattttctct tggtgggctg
     1381 gggatgaagg ccctgccctg gaggcactca tcagactgta taaacagaag tatcctggag
     1441 tagaggtgat aaacgccact gtcaccggag gtgccggtgt aaatgctcgc gctgtcctta
     1501 agacaaggat gcttggtggt gatcctcctg atacatttca agttgcagca gggatggagt
     1561 tgataggcac atgggtggtg gctaacagaa tggaggatct cagcgccttg tttcgacaag
     1621 agggttggct tcaagccttc cccaagggcc ttattgatct gattagttat aaaggtggta
     1681 tatggagtgt gcccgttaat attcacaggt caaacgtaat gtggtatttg cctgcaaagc
     1741 taaaagagtg gggagttaac cccccacgga cttgggatga gtttcttgcc acctgtcaaa
     1801 cacttaaaca aaagggtctg gaagctccac tggctctcgg ggaaaattgg acacaacaac
     1861 acctctggga gagtgtcgcc ttggctgtat tggggcctga tgattggaat aatctgtgga
     1921 atgggaagct gaagttcaca gaccctaaag cagttagggc atgggaagtc tttggccgcg
     1981 ttttggactg tgccaacaag gatgcagcag gcttgtcttg gcagcaagct gtcgatcggg
     2041 tagtacaggg gaaagccgct tttaacgtaa tgggcgactg ggcagcagga tatatgacta
     2101 ccactctcaa gttgaagcct ggtacagatt tcgcatgggc accaagtccc gggacacagg
     2161 gcgtctttat gatggtttcc gactctttcg gacttcccaa aggagccaaa aatcgacaaa
     2221 acgctatcaa ctggcttcga ttggtcggct ctaaagaagg tcaagataca ttcaacccac
     2281 tgaaagggtc catcgctgct cggctcgata gcgacccaag caagtatccg tgtaacgtct
     2341 atatcaccgc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaacg
     2401 tggaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     2461 gccccgtgct gctgcccgac aaccactacc tggtgaccca gtccgtgctg agcaaagacc
     2521 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     2581 tcggcatgga cgagctgtac aagggcggta ccggagggag catgagcaag ggcgaggagc
     2641 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     2701 tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca
     2761 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     2821 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     2881 ccatgcccga aggctacgtc caggagcgca ccatcagctt caaggacgac ggcacctaca
     2941 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     3001 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactttaaca
     3061 accccaacgc atacggacag agtgcaatgc gagattggcg gagtaatcgc atcgttggct
     3121 ccctggtagc aggagcagtg gcccctgaaa gctccatgtc acaattcgga acagtaatgg
     3181 aaattttcct gcaaacacgc aacccacaag cagcagcaaa tgcagcacaa gctattgctg
     3241 accaagttgg cctgggtcgc cttggtcagc tgcaggtcga cgaacaaaaa ctcatctcag
     3301 aagaggatct gaatgctgtg ggccaggaca cgcaggaggt catcgtggtg ccacactcct
     3361 tgccctttaa ggtggtggtg atctcagcca tcctggccct ggtggtgctc accatcatct
     3421 cccttatcat cctcatcatg ctttggcaga agaagccacg ttagaagctt gctagtataa
     3481 cttcgtatag catacattat acgaagttat gggtcgatgg tgatgcttgg caattcgggt
     3541 cgatggtgaa gcattggtct tcctattttg atataacttc gtataggata ctttatacga
     3601 agttatgggg atgatccaat tctttgccaa aatgatgaga cagcacaata accagcacgt
     3661 tgcccaggag ctgtaggaaa aagaagaagg catgaacatg gttagcagag gctctagagc
     3721 cgccggtcac acgccagaag ccgaaccccg ccctgccccg tcccccccga aggcagccgt
     3781 ccccccgcgg acagccccga gccgctcccc ccgcacaaag ggccctcccg gagcccctca
     3841 aggctttcac gcagccacag aaaagaaaca agccgtcatt aaaccaagcg ctaattacag
     3901 cccggaggag aagggccgtc ccgcccgctc acctgtggga gtaacgcggt cagtcagagc
     3961 cggggcgggc ggcgcgaggc ggcggcggag cggggcacgg ggcgaaggca gcgcgcagcg
     4021 actcccgccc gccgcgcgct tcgcttttta tagggccgcc gccgccgccg cctcgccata
     4081 aaaggaaact ttcggagcgc gccgctctga ttggctgccg ccgcacctct ccgcctcgcc
     4141 ccgccccgcc cctcgccccg ccccgccccg cctggcgcgc gccccccccc cccccccgcc
     4201 cccatcgctg cacaaaataa ttaaaaaata aataaataca aaattggggg tggggagggg
     4261 ggggagatgg ggagagtgaa gcagaacgtg gggctcacct cgaccatggt aatagcgatg
     4321 actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa
     4381 tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac
     4441 acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg
     4501 aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tggtactctt
     4561 cctagagcat ggctacgtag ataagtagca tggcgggtta atcattaact acaaggaacc
     4621 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     4681 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     4741 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     4801 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4861 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4921 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4981 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     5041 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5101 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5161 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5221 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     5281 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5341 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5401 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5461 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5521 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5581 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5641 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5701 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5761 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5821 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5881 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5941 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6001 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6061 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6121 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6181 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6241 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6301 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6361 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6421 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6481 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6541 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6601 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6661 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6721 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6781 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6841 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6901 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6961 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7021 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7081 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     7141 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     7201 g
//
