LOCUS       sequence-494291-        7199 bp    DNA     circular SYN 02-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 7199)
  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 7199)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 2, 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.hSynap.(mem).iGlucoSnFR2.mRuby3
FEATURES             Location/Qualifiers
     source          1..7199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(237..825)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(317..336)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(996..1787,1788..1856))
                     /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     1619..1638
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1857..1961)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1987..2442)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2124..2145)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2336..2355
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2514..2643
                     /label=AAV2 ITR
     promoter        2719..3166
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3195..3204
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3201..3263
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     primer_bind     4457..4478
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4581..4602)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             5199..5228
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5232..5378
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    6150..6738
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6203..6223)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6621..6632)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6741..6757)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6742..6758)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    6787..6908
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6798..6817)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6852..6871
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   7070..7199
                     /label=AAV2 ITR
ORIGIN
        1 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
       61 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      121 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      181 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      241 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      301 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      361 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
      421 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
      481 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
      541 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
      601 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
      661 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
      721 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
      781 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
      841 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
      901 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
      961 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     1021 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     1081 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     1141 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     1201 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     1261 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     1321 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     1381 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     1441 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     1501 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     1561 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     1621 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     1681 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     1741 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     1801 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     1861 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     1921 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     1981 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     2041 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     2101 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     2161 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     2221 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     2281 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2341 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2401 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
     2461 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgcgcg
     2521 ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc
     2581 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
     2641 ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga
     2701 tccagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc ggggtggggg
     2761 tgcctacctg acgaccggcc ccgacccact ggacaagcac ccaaccccca ttccccaaat
     2821 tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg cgtgcgcact
     2881 gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc gccaccgccg
     2941 cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc cccttcccgg
     3001 ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg cgaggcgcga
     3061 gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag cgctgcctca
     3121 gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcca attcaagctg
     3181 ctagcaagga tcccgccacc atggagacag acacactcct gctatgggta ctgctgctct
     3241 gggttccagg ttccactggt gacagatcta agctggagat tttctcttgg tgggctgggg
     3301 atgaaggccc tgccctggag gcactcatca gactgtataa acagaagtat cctggagtag
     3361 aggtgataaa cgccactgtc accggaggtg ccggtgtaaa tgctcgcgct gtccttaaga
     3421 caaggatgct tggtggtgat cctcctgata catttcaagt tgcagcaggg atggagttga
     3481 taggcacatg ggtggtggct aacagaatgg aggatctcag cgccttgttt cgacaagagg
     3541 gttggcttca agccttcccc aagggcctta ttgatctgat tagttataaa ggtggtatat
     3601 ggagtgtgcc cgttaatatt cacaggtcaa acgtaatgtg gtatttgcct gcaaagctaa
     3661 aagagtgggg agttaacccc ccacggactt gggatgagtt tcttgccacc tgtcaaacac
     3721 ttaaacaaaa gggtctggaa gctccactgg ctctcgggga aaattggaca caacaacacc
     3781 tctgggagag tgtcgccttg gctgtattgg ggcctgatga ttggaataat ctgtggaatg
     3841 ggaagctgaa gttcacagac cctaaagcag ttagggcatg ggaagtcttt ggccgcgttt
     3901 tggactgtgc caacaaggat gcagcaggct tgtcttggca gcaagctgtc gatcgggtag
     3961 tacaggggaa agccgctttt aacgtaatgg gcgactgggc agcaggatat atgactacca
     4021 ctctcaagtt gaagcctggt acagatttcg catgggcacc aagtcccggg acacagggcg
     4081 tctttatgat ggtttccgac tctttcggac ttcccaaagg agccaaaaat cgacaaaacg
     4141 ctatcaactg gcttcgattg gtcggctcta aagaaggtca agatacattc aacccactga
     4201 aagggtccat cgctgctcgg ctcgatagcg acccaagcaa gtatccgtgt aacgtctata
     4261 tcaccgccga caagcagaag aacggcatca aggcgaactt caagatccgc cacaacgtgg
     4321 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     4381 ccgtgctgct gcccgacaac cactacctgg tgacccagtc cgtgctgagc aaagacccca
     4441 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     4501 gcatggacga gctgtacaag ggcggtaccg gaggaagcat gagcaagggc gaggagctgt
     4561 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
     4621 gcgtgcgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg aagttcatct
     4681 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
     4741 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
     4801 tgcccgaagg ctacgtccag gagcgcacca tcagcttcaa ggacgacggc acctacaaga
     4861 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
     4921 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tttaacaacc
     4981 ccaacgcata cggacagagt gcaatgcgag attggcggag taatcgcatc gttggctccc
     5041 tggtagcagg agcagtggcc cctgaaagct ccatgtcaca attcggaaca gtaatggaaa
     5101 ttttcctgca aacacgcaac ccacaagcag cagcaaatgc agcacaagct attgctgacc
     5161 aagttggcct gggtcgcctt ggtcagctgc aggtcgacga acaaaaactc atctcagaag
     5221 aggatctgaa tgctgtgggc caggacacgc aggaggtcat cgtggtgcca cactccttgc
     5281 cctttaaggt ggtggtgatc tcagccatcc tggccctggt ggtgctcacc atcatctccc
     5341 ttatcatcct catcatgctt tggcagaaga agccacgttc tagaggctcc ggaaatgctg
     5401 tgggccagga cacgcaggag cgtatggtgt ctaagggcga agagctgatc aaggaaaata
     5461 tgcgtatgaa ggtggtcatg gaaggttcgg tcaacggcca ccaattcaaa tgcacaggtg
     5521 aaggagaagg caggccgtac gagggagtgc aaaccatgag gatcaaagtc atcgagggag
     5581 gacccctgcc atttgccttt gacattcttg ccacgtcgtt catgtatggc agccgtactt
     5641 ttatcaagta cccggcagac attcctgatt tctttaaaca gtcctttcct gagggtttta
     5701 cttgggaaag agttacgaga tacgaagatg gtggagtcgt caccgtcacg caggacacca
     5761 gccttgagga tggcgaactc gtttacaacg tcaaagtcag aggggtaaac tttccctcca
     5821 atggtcccgt gatgcagaag aagaccaagg gttgggagcc taatacagag atgatgtatc
     5881 cagcagatgg tggtctgagg ggatacactg atattgcact gaaagttgat ggtggtggcc
     5941 atctgcattg caatttcgta acaacttaca ggtcaaaaaa gaccgtcggg aacatcaaga
     6001 tgcccggtgt ccatgccgtt gatcaccgcc tggaaaggat agaggaaagt gacaatgaaa
     6061 cgtacgtagt acaacgcgaa gtcgcagttg ccaagtactc caatcttggt ggtggtatgg
     6121 atgaacttta taagtagaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     6181 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     6241 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     6301 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     6361 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     6421 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     6481 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     6541 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     6601 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     6661 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     6721 tttgggccgc ctccccgcat cgataccgtc gacctcgacc cgggcggccg cttcgagcag
     6781 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     6841 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     6901 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     6961 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta
     7021 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     7081 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     7141 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcag
//
