LOCUS       sequence-486288-        7403 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasma membrane targeted expression of green glucose sensor with 
            non-responsive mRuby3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7403)
  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 7403)
  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.GFAP.(mem).iGlucoSnFR2.mRuby3
FEATURES             Location/Qualifiers
     source          1..7403
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(17..472)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(154..175)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     366..385
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   544..673
                     /label=AAV2 ITR
     regulatory      1438..1447
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1444..1503
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     2700..2721
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2824..2845)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             3442..3471
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3475..3621
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    4405..4993
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4458..4478)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4876..4887)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    5021..5142
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5032..5051)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5086..5105
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5304..5433
                     /label=AAV2 ITR
     rep_origin      complement(5670..6258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5750..5769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6429..7220,7221..7289))
                     /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"
     promoter        complement(7290..7394)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
       61 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
      121 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
      181 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
      241 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
      301 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
      361 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
      421 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
      481 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
      541 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
      601 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
      661 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
      721 tctaggaaga tcgtaggtta attaataaca tatcctggtg tggagtaggg gacgctgctc
      781 tgacagaggc tcgggggcct gagctggctc tgtgagctgg ggaggaggca gacagccagg
      841 cctttgtctg caagcagacc tggcagcatt gggctggccg ccccccaggg cctcctcttc
      901 atgcccagtg aatgactcac cttggcacag acacaatgtt cggggtgggc acagtgcctg
      961 cttcccgccg caccccagcc cccctcaaat gccttccgag aagcccattg agcagggggc
     1021 ttgcattgca ccccagcctg acagcctggc atcttgggat aaaagcagca cagcccccta
     1081 ggggctgccc ttgctgtgtg gcgccaccgg cggtggagaa caaggctcta ttcagcctgt
     1141 gcccaggaaa ggggatcagg ggatgcccag gcatggacag tgggtggcag ggggggagag
     1201 gagggctgtc tgcttcccag aagtccaagg acacaaatgg gtgaggggag agctctcccc
     1261 atagctgggc tgcggcccaa ccccaccccc tcaggctatg ccagggggtg ttgccagggg
     1321 cacccgggca tcgccagtct agcccactcc ttcataaagc cctcgcatcc caggagcgag
     1381 cagagccaga gcaggttgga gaggagacgc atcacctccg ctgctcgcgg ggatcccgcc
     1441 accatggaga cagacacact cctgctatgg gtactgctgc tctgggttcc aggttccact
     1501 ggtgacagat ctaagctgga gattttctct tggtgggctg gggatgaagg ccctgccctg
     1561 gaggcactca tcagactgta taaacagaag tatcctggag tagaggtgat aaacgccact
     1621 gtcaccggag gtgccggtgt aaatgctcgc gctgtcctta agacaaggat gcttggtggt
     1681 gatcctcctg atacatttca agttgcagca gggatggagt tgataggcac atgggtggtg
     1741 gctaacagaa tggaggatct cagcgccttg tttcgacaag agggttggct tcaagccttc
     1801 cccaagggcc ttattgatct gattagttat aaaggtggta tatggagtgt gcccgttaat
     1861 attcacaggt caaacgtaat gtggtatttg cctgcaaagc taaaagagtg gggagttaac
     1921 cccccacgga cttgggatga gtttcttgcc acctgtcaaa cacttaaaca aaagggtctg
     1981 gaagctccac tggctctcgg ggaaaattgg acacaacaac acctctggga gagtgtcgcc
     2041 ttggctgtat tggggcctga tgattggaat aatctgtgga atgggaagct gaagttcaca
     2101 gaccctaaag cagttagggc atgggaagtc tttggccgcg ttttggactg tgccaacaag
     2161 gatgcagcag gcttgtcttg gcagcaagct gtcgatcggg tagtacaggg gaaagccgct
     2221 tttaacgtaa tgggcgactg ggcagcagga tatatgacta ccactctcaa gttgaagcct
     2281 ggtacagatt tcgcatgggc accaagtccc gggacacagg gcgtctttat gatggtttcc
     2341 gactctttcg gacttcccaa aggagccaaa aatcgacaaa acgctatcaa ctggcttcga
     2401 ttggtcggct ctaaagaagg tcaagataca ttcaacccac tgaaagggtc catcgctgct
     2461 cggctcgata gcgacccaag caagtatccg tgtaacgtct atatcaccgc cgacaagcag
     2521 aagaacggca tcaaggcgaa cttcaagatc cgccacaacg tggaggacgg cagcgtgcag
     2581 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2641 aaccactacc tggtgaccca gtccgtgctg agcaaagacc ccaacgagaa gcgcgatcac
     2701 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2761 aagggcggta ccggaggaag catgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
     2821 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgcg cggcgagggc
     2881 gagggcgatg ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
     2941 cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc
     3001 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     3061 caggagcgca ccatcagctt caaggacgac ggcacctaca agacccgcgc cgaggtgaag
     3121 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     3181 ggcaacatcc tggggcacaa gctggagtac aactttaaca accccaacgc atacggacag
     3241 agtgcaatgc gagattggcg gagtaatcgc atcgttggct ccctggtagc aggagcagtg
     3301 gcccctgaaa gctccatgtc acaattcgga acagtaatgg aaattttcct gcaaacacgc
     3361 aacccacaag cagcagcaaa tgcagcacaa gctattgctg accaagttgg cctgggtcgc
     3421 cttggtcagc tgcaggtcga cgaacaaaaa ctcatctcag aagaggatct gaatgctgtg
     3481 ggccaggaca cgcaggaggt catcgtggtg ccacactcct tgccctttaa ggtggtggtg
     3541 atctcagcca tcctggccct ggtggtgctc accatcatct cccttatcat cctcatcatg
     3601 ctttggcaga agaagccacg ttctagaggc tccggaaatg ctgtgggcca ggacacgcag
     3661 gagcgtatgg tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat gaaggtggtc
     3721 atggaaggtt cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga aggcaggccg
     3781 tacgagggag tgcaaaccat gaggatcaaa gtcatcgagg gaggacccct gccatttgcc
     3841 tttgacattc ttgccacgtc gttcatgtat ggcagccgta cttttatcaa gtacccggca
     3901 gacattcctg atttctttaa acagtccttt cctgagggtt ttacttggga aagagttacg
     3961 agatacgaag atggtggagt cgtcaccgtc acgcaggaca ccagccttga ggatggcgaa
     4021 ctcgtttaca acgtcaaagt cagaggggta aactttccct ccaatggtcc cgtgatgcag
     4081 aagaagacca agggttggga gcctaataca gagatgatgt atccagcaga tggtggtctg
     4141 aggggataca ctgatattgc actgaaagtt gatggtggtg gccatctgca ttgcaatttc
     4201 gtaacaactt acaggtcaaa aaagaccgtc gggaacatca agatgcccgg tgtccatgcc
     4261 gttgatcacc gcctggaaag gatagaggaa agtgacaatg aaacgtacgt agtacaacgc
     4321 gaagtcgcag ttgccaagta ctccaatctt ggtggtggta tggatgaact ttataagtag
     4381 aagcttttcg aattagctta tcataatcaa cctctggatt acaaaatttg tgaaagattg
     4441 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     4501 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     4561 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     4621 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     4681 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     4741 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     4801 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     4861 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     4921 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     4981 gccgcctccc cgcatcggag gccgcttcga gcagacatga taagatacat tgatgagttt
     5041 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     5101 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     5161 cattttatgt ttcaggttca gggggagatg tgggaggttt tttaaagcaa gtaaaacctc
     5221 tacaaatgtg gtaaaatcga taaggatctt cctagagcat ggctacgtag ataagtagca
     5281 tggcgggtta atcattaact acaaggaacc cctagtgatg gagttggcca ctccctctct
     5341 gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc
     5401 ccgggcggcc tcagtgagcg agcgagcgcg cagctgcatt aatgaatcgg ccaacgcgcg
     5461 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     5521 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     5581 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5641 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5701 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5761 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5821 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5881 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5941 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6001 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6061 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6121 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6181 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6241 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6301 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6361 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6421 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6481 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6541 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6601 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6661 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6721 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6781 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6841 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6901 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6961 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7021 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7081 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7141 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7201 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7261 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7321 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7381 ataggggttc cgcgcacatt tcc
//
