LOCUS       sequence-487084-        6984 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic 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 6984)
  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 6984)
  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.hSynap.(cyto).iGlucoSnFR2.mRuby3
FEATURES             Location/Qualifiers
     source          1..6984
                     /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"
     primer_bind     4400..4421
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4524..4545)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             5895..5912
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    5935..6523
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5988..6008)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6406..6417)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6526..6542)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6527..6543)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    6572..6693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6583..6602)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6637..6656
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6855..6984
                     /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 atgggcagat ctaagctgga gattttctct tggtgggctg
     3241 gggatgaagg ccctgccctg gaggcactca tcagactgta taaacagaag tatcctggag
     3301 tagaggtgat aaacgccact gtcaccggag gtgccggtgt aaatgctcgc gctgtcctta
     3361 agacaaggat gcttggtggt gatcctcctg atacatttca agttgcagca gggatggagt
     3421 tgataggcac atgggtggtg gctaacagaa tggaggatct cagcgccttg tttcgacaag
     3481 agggttggct tcaagccttc cccaagggcc ttattgatct gattagttat aaaggtggta
     3541 tatggagtgt gcccgttaat attcacaggt caaacgtaat gtggtatttg cctgcaaagc
     3601 taaaagagtg gggagttaac cccccacgga cttgggatga gtttcttgcc acctgtcaaa
     3661 cacttaaaca aaagggtctg gaagctccac tggctctcgg ggaaaattgg acacaacaac
     3721 acctctggga gagtgtcgcc ttggctgtat tggggcctga tgattggaat aatctgtgga
     3781 atgggaagct gaagttcaca gaccctaaag cagttagggc atgggaagtc tttggccgcg
     3841 ttttggactg tgccaacaag gatgcagcag gcttgtcttg gcagcaagct gtcgatcggg
     3901 tagtacaggg gaaagccgct tttaacgtaa tgggcgactg ggcagcagga tatatgacta
     3961 ccactctcaa gttgaagcct ggtacagatt tcgcatgggc accaagtccc gggacacagg
     4021 gcgtctttat gatggtttcc gactctttcg gacttcccaa aggagccaaa aatcgacaaa
     4081 acgctatcaa ctggcttcga ttggtcggct ctaaagaagg tcaagataca ttcaacccac
     4141 tgaaagggtc catcgctgct cggctcgata gcgacccaag caagtatccg tgtaacgtct
     4201 atatcaccgc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaacg
     4261 tggaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     4321 gccccgtgct gctgcccgac aaccactacc tggtgaccca gtccgtgctg agcaaagacc
     4381 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     4441 tcggcatgga cgagctgtac aagggcggta ccggagggag catgagcaag ggcgaggagc
     4501 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     4561 tcagcgtgcg cggcgagggc gagggcgatg ccaccaacgg caagctgacc ctgaagttca
     4621 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     4681 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     4741 ccatgcccga aggctacgtc caggagcgca ccatcagctt caaggacgac ggcacctaca
     4801 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     4861 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactttaaca
     4921 accccaacgc atacggacag agtgcaatgc gagattggcg gagtaatcgc atcgttggct
     4981 ccctggtagc aggagcagtg gcccctgaaa gctccatgtc acaattcgga acagtaatgg
     5041 aaattttcct gcaaacacgc aacccacaag cagcagcaaa tgcagcacaa gctattgctg
     5101 accaagttgg cctgggtcgc cttggtcagc tgcagtctac cggctccgga aatgctgtgg
     5161 gccaggacac gcaggagcgt atggtgtcta agggcgaaga gctgatcaag gaaaatatgc
     5221 gtatgaaggt ggtcatggaa ggttcggtca acggccacca attcaaatgc acaggtgaag
     5281 gagaaggcag gccgtacgag ggagtgcaaa ccatgaggat caaagtcatc gagggaggac
     5341 ccctgccatt tgcctttgac attcttgcca cgtcgttcat gtatggcagc cgtactttta
     5401 tcaagtaccc ggcagacatt cctgatttct ttaaacagtc ctttcctgag ggttttactt
     5461 gggaaagagt tacgagatac gaagatggtg gagtcgtcac cgtcacgcag gacaccagcc
     5521 ttgaggatgg cgaactcgtt tacaacgtca aagtcagagg ggtaaacttt ccctccaatg
     5581 gtcccgtgat gcagaagaag accaagggtt gggagcctaa tacagagatg atgtatccag
     5641 cagatggtgg tctgagggga tacactgata ttgcactgaa agttgatggt ggtggccatc
     5701 tgcattgcaa tttcgtaaca acttacaggt caaaaaagac cgtcgggaac atcaagatgc
     5761 ccggtgtcca tgccgttgat caccgcctgg aaaggataga ggaaagtgac aatgaaacgt
     5821 acgtagtaca acgcgaagtc gcagttgcca agtactccaa tcttggtggt ggtatggatg
     5881 aactttataa gtcccatcac catcaccacc attaagaatt cgaagcttat cgataatcaa
     5941 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     6001 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     6061 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     6121 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     6181 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     6241 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     6301 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     6361 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     6421 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     6481 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgtcgacct
     6541 cgacccgggc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc
     6601 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     6661 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     6721 tttcaggttc agggggagat gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt
     6781 ggtaaaatcg ataaggatct tcctagagca tggctacgta gataagtagc atggcgggtt
     6841 aatcattaac tacaaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg
     6901 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc
     6961 ctcagtgagc gagcgagcgc gcag
//
