LOCUS       sequence-487025-        7957 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 7957)
  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 7957)
  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.mRuby3
FEATURES             Location/Qualifiers
     source          1..7957
                     /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(4233..4266)
                     /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(4329..4362)
                     /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(4400..4419)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4481..4503)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(4782..5059)
                     /label=chicken beta-actin promoter
     repeat_region   5370..5499
                     /label=AAV2 ITR
     rep_origin      5571..6026
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5658..5677)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5868..5889
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6052..6156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6157..6225,6226..7017)
                     /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(6375..6394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7188..7776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7677..7696
                     /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 ccggaggaag 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 ttctagaggc tccggaaatg
     3481 ctgtgggcca ggacacgcag gagcgtatgg tgtctaaggg cgaagagctg atcaaggaaa
     3541 atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg ccaccaattc aaatgcacag
     3601 gtgaaggaga aggcaggccg tacgagggag tgcaaaccat gaggatcaaa gtcatcgagg
     3661 gaggacccct gccatttgcc tttgacattc ttgccacgtc gttcatgtat ggcagccgta
     3721 cttttatcaa gtacccggca gacattcctg atttctttaa acagtccttt cctgagggtt
     3781 ttacttggga aagagttacg agatacgaag atggtggagt cgtcaccgtc acgcaggaca
     3841 ccagccttga ggatggcgaa ctcgtttaca acgtcaaagt cagaggggta aactttccct
     3901 ccaatggtcc cgtgatgcag aagaagacca agggttggga gcctaataca gagatgatgt
     3961 atccagcaga tggtggtctg aggggataca ctgatattgc actgaaagtt gatggtggtg
     4021 gccatctgca ttgcaatttc gtaacaactt acaggtcaaa aaagaccgtc gggaacatca
     4081 agatgcccgg tgtccatgcc gttgatcacc gcctggaaag gatagaggaa agtgacaatg
     4141 aaacgtacgt agtacaacgc gaagtcgcag ttgccaagta ctccaatctt ggtggtggta
     4201 tggatgaact ttataagtag aagcttgcta gtataacttc gtatagcata cattatacga
     4261 agttatgggt cgatggtgat gcttggcaat tcgggtcgat ggtgaagcat tggtcttcct
     4321 attttgatat aacttcgtat aggatacttt atacgaagtt atggggatga tccaattctt
     4381 tgccaaaatg atgagacagc acaataacca gcacgttgcc caggagctgt aggaaaaaga
     4441 agaaggcatg aacatggtta gcagaggctc tagagccgcc ggtcacacgc cagaagccga
     4501 accccgccct gccccgtccc ccccgaaggc agccgtcccc ccgcggacag ccccgagccg
     4561 ctccccccgc acaaagggcc ctcccggagc ccctcaaggc tttcacgcag ccacagaaaa
     4621 gaaacaagcc gtcattaaac caagcgctaa ttacagcccg gaggagaagg gccgtcccgc
     4681 ccgctcacct gtgggagtaa cgcggtcagt cagagccggg gcgggcggcg cgaggcggcg
     4741 gcggagcggg gcacggggcg aaggcagcgc gcagcgactc ccgcccgccg cgcgcttcgc
     4801 tttttatagg gccgccgccg ccgccgcctc gccataaaag gaaactttcg gagcgcgccg
     4861 ctctgattgg ctgccgccgc acctctccgc ctcgccccgc cccgcccctc gccccgcccc
     4921 gccccgcctg gcgcgcgccc cccccccccc cccgccccca tcgctgcaca aaataattaa
     4981 aaaataaata aatacaaaat tgggggtggg gagggggggg agatggggag agtgaagcag
     5041 aacgtggggc tcacctcgac catggtaata gcgatgacta atacgtagat gtactgccaa
     5101 gtaggaaagt cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca
     5161 ttgacgtcaa tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca
     5221 gtttaccgta aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg
     5281 ggaacatacg tcattattga cgtcaatggt actcttccta gagcatggct acgtagataa
     5341 gtagcatggc gggttaatca ttaactacaa ggaaccccta gtgatggagt tggccactcc
     5401 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg
     5461 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tggcgtaata gcgaagaggc
     5521 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg
     5581 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     5641 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     5701 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     5761 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     5821 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     5881 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     5941 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     6001 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     6061 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6121 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6181 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6241 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6301 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6361 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6421 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6481 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6541 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6601 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6661 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6721 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6781 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6841 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6901 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6961 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7021 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7081 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7141 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7201 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7261 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7321 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7381 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7441 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7501 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7561 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7621 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7681 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7741 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7801 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7861 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7921 accgagcgca gcgagtcagt gagcgaggaa gcggaag
//
