LOCUS       sequence-487005-        7777 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 7777)
  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 7777)
  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.hSynapFLEX.(mem).iGlucoSnFR2.mRuby3
FEATURES             Location/Qualifiers
     source          1..7777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      69..524
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(156..175)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     366..387
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        550..654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(655..723,724..1515)
                     /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(873..892)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1686..2274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2175..2194
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2511..2640
                     /label=AAV2 ITR
     polyA_signal    2802..2923
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2839..2858)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2893..2912
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3001..3589
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3107..3118
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3516..3536
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3661..3694
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3765..3798
                     /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      3823..3832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3829..3888
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     primer_bind     5085..5106
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5209..5230)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             5827..5856
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5860..6006
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     protein_bind    complement(6778..6811)
                     /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(6874..6907)
                     /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(6918..6937)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          6920..7052
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   7645..7774
                     /label=AAV2 ITR
ORIGIN
        1 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
       61 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
      121 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
      181 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      241 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
      301 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      361 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
      421 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
      481 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
      541 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      601 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      661 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      721 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      781 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      841 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      901 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
      961 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1021 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1081 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1141 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1201 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1261 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1321 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1381 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1441 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1501 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1561 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1621 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1681 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1741 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1801 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1861 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1921 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1981 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2041 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2101 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2161 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2221 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2281 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2341 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2401 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2461 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctgcgcgctc
     2521 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg
     2581 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     2641 tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct aggaagatcc
     2701 ttatcgattt taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacatctcc
     2761 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2821 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2881 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgctcgaagc
     2941 ggccgcccgg gtcgactcta gatgcatgct cgagcggccg cccgggtcga cggtatcgat
     3001 gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga agacgcggaa
     3061 gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg agggccgaag
     3121 ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacacaggcg agcagccaag
     3181 gaaaggacga tgatttcccc gacaacacca cggaattgtc agtgcccaac agccgagccc
     3241 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata gggaggggga
     3301 aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca gtgggggttg
     3361 cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac aactcctcat
     3421 aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
     3481 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
     3541 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta tcgataagct
     3601 tctcgagtgg atcatctgca gaattcgccc ttttcagtgt ttcgaacctc ttcgagggac
     3661 ataacttcgt ataatgtatg ctatacgaag ttatgggtcg atggtgagat ctggactaga
     3721 gggtcgatgg tgatgcttgg atcaaaatag gaagaatccc aatgataact tcgtataaag
     3781 tatcctatac gaagttatac tagtggtacc accggtgata tcgccaccat ggagacagac
     3841 acactcctgc tatgggtact gctgctctgg gttccaggtt ccactggtga cagatctaag
     3901 ctggagattt tctcttggtg ggctggggat gaaggccctg ccctggaggc actcatcaga
     3961 ctgtataaac agaagtatcc tggagtagag gtgataaacg ccactgtcac cggaggtgcc
     4021 ggtgtaaatg ctcgcgctgt ccttaagaca aggatgcttg gtggtgatcc tcctgataca
     4081 tttcaagttg cagcagggat ggagttgata ggcacatggg tggtggctaa cagaatggag
     4141 gatctcagcg ccttgtttcg acaagagggt tggcttcaag ccttccccaa gggccttatt
     4201 gatctgatta gttataaagg tggtatatgg agtgtgcccg ttaatattca caggtcaaac
     4261 gtaatgtggt atttgcctgc aaagctaaaa gagtggggag ttaacccccc acggacttgg
     4321 gatgagtttc ttgccacctg tcaaacactt aaacaaaagg gtctggaagc tccactggct
     4381 ctcggggaaa attggacaca acaacacctc tgggagagtg tcgccttggc tgtattgggg
     4441 cctgatgatt ggaataatct gtggaatggg aagctgaagt tcacagaccc taaagcagtt
     4501 agggcatggg aagtctttgg ccgcgttttg gactgtgcca acaaggatgc agcaggcttg
     4561 tcttggcagc aagctgtcga tcgggtagta caggggaaag ccgcttttaa cgtaatgggc
     4621 gactgggcag caggatatat gactaccact ctcaagttga agcctggtac agatttcgca
     4681 tgggcaccaa gtcccgggac acagggcgtc tttatgatgg tttccgactc tttcggactt
     4741 cccaaaggag ccaaaaatcg acaaaacgct atcaactggc ttcgattggt cggctctaaa
     4801 gaaggtcaag atacattcaa cccactgaaa gggtccatcg ctgctcggct cgatagcgac
     4861 ccaagcaagt atccgtgtaa cgtctatatc accgccgaca agcagaagaa cggcatcaag
     4921 gcgaacttca agatccgcca caacgtggag gacggcagcg tgcagctcgc cgaccactac
     4981 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctggtg
     5041 acccagtccg tgctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     5101 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg cggtaccgga
     5161 ggaagcatga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     5221 gacggcgacg taaacggcca caagttcagc gtgcgcggcg agggcgaggg cgatgccacc
     5281 aacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     5341 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     5401 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     5461 agcttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     5521 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     5581 cacaagctgg agtacaactt taacaacccc aacgcatacg gacagagtgc aatgcgagat
     5641 tggcggagta atcgcatcgt tggctccctg gtagcaggag cagtggcccc tgaaagctcc
     5701 atgtcacaat tcggaacagt aatggaaatt ttcctgcaaa cacgcaaccc acaagcagca
     5761 gcaaatgcag cacaagctat tgctgaccaa gttggcctgg gtcgccttgg tcagctgcag
     5821 gtcgacgaac aaaaactcat ctcagaagag gatctgaatg ctgtgggcca ggacacgcag
     5881 gaggtcatcg tggtgccaca ctccttgccc tttaaggtgg tggtgatctc agccatcctg
     5941 gccctggtgg tgctcaccat catctccctt atcatcctca tcatgctttg gcagaagaag
     6001 ccacgttcta gaggctccgg aaatgctgtg ggccaggaca cgcaggagcg tatggtgtct
     6061 aagggcgaag agctgatcaa ggaaaatatg cgtatgaagg tggtcatgga aggttcggtc
     6121 aacggccacc aattcaaatg cacaggtgaa ggagaaggca ggccgtacga gggagtgcaa
     6181 accatgagga tcaaagtcat cgagggagga cccctgccat ttgcctttga cattcttgcc
     6241 acgtcgttca tgtatggcag ccgtactttt atcaagtacc cggcagacat tcctgatttc
     6301 tttaaacagt cctttcctga gggttttact tgggaaagag ttacgagata cgaagatggt
     6361 ggagtcgtca ccgtcacgca ggacaccagc cttgaggatg gcgaactcgt ttacaacgtc
     6421 aaagtcagag gggtaaactt tccctccaat ggtcccgtga tgcagaagaa gaccaagggt
     6481 tgggagccta atacagagat gatgtatcca gcagatggtg gtctgagggg atacactgat
     6541 attgcactga aagttgatgg tggtggccat ctgcattgca atttcgtaac aacttacagg
     6601 tcaaaaaaga ccgtcgggaa catcaagatg cccggtgtcc atgccgttga tcaccgcctg
     6661 gaaaggatag aggaaagtga caatgaaacg tacgtagtac aacgcgaagt cgcagttgcc
     6721 aagtactcca atcttggtgg tggtatggat gaactttata agtaagctag cactagtata
     6781 acttcgtata gcatacatta tacgaagtta tgtgtcgatg gtgatgcttg gcaattcggg
     6841 tcgatggtga agcattggtc ttcctatttt gatataactt cgtataggat actttatacg
     6901 aagttatggg gatcatcacc tgtggagaga aaggcaaagt ggatgtcagt aagaccaata
     6961 ggtgcctatc agaaacgcaa gagtcttctc tgtctcgaca agcccagttt ctattggtct
     7021 ccttaaacct gtcttgtaac cttgatactt acctgatcca acggctgcga cttggggcag
     7081 ggggtcctag gggtggtctg gccaggagcc gcgggggcgg actgcgcggt gcccaggagc
     7141 acagctgcgc tctcaggcac gacacgactc ctccgctgcc caccgcagac tgaggcagcg
     7201 ctgagtcgcc ggcgccgcag cgcagatggt cgcgcccgtg cccccctatc tcgcgcctcg
     7261 cgtggtgcgg tccggctggg ccggcggcgg cgcggacgcg accaaggtgg ccgggaaggg
     7321 gagtttgcgg gggaccggcg agtgacgtca gcgcgccttc agtgctgagg cggcggtggc
     7381 gcgcgccgcc aggcgggggc gaaggcactg tccgcggtgc tgaagctggc agtgcgcacg
     7441 cgcctcgccg catcctgttt cccctccccc tctctgatag gggatgcgca atttggggaa
     7501 tgggggttgg gtgcttgtcc agtgggtcgg ggtcggtcgt caatttaaat actgaaaagg
     7561 gcgaattcca gcacactgga gctcggatct tcctagagca tggctacgta gataagtagc
     7621 atggcgggtt aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc
     7681 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg
     7741 cccgggcggc ctcagtgagc gagcgagcgc gcagctg
//
