LOCUS       sequence-486283-        7188 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 7188)
  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 7188)
  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.(cyto).iGlucoSnFR2.mRuby3
FEATURES             Location/Qualifiers
     source          1..7188
                     /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"
     primer_bind     2643..2664
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2767..2788)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             4138..4155
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    4190..4778
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4243..4263)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4661..4672)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4806..4927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4817..4836)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4871..4890
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5089..5218
                     /label=AAV2 ITR
     rep_origin      complement(5455..6043)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5535..5554)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6214..7005,7006..7074))
                     /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(7075..7179)
                     /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 accatgggca gatctaagct ggagattttc tcttggtggg ctggggatga aggccctgcc
     1501 ctggaggcac tcatcagact gtataaacag aagtatcctg gagtagaggt gataaacgcc
     1561 actgtcaccg gaggtgccgg tgtaaatgct cgcgctgtcc ttaagacaag gatgcttggt
     1621 ggtgatcctc ctgatacatt tcaagttgca gcagggatgg agttgatagg cacatgggtg
     1681 gtggctaaca gaatggagga tctcagcgcc ttgtttcgac aagagggttg gcttcaagcc
     1741 ttccccaagg gccttattga tctgattagt tataaaggtg gtatatggag tgtgcccgtt
     1801 aatattcaca ggtcaaacgt aatgtggtat ttgcctgcaa agctaaaaga gtggggagtt
     1861 aaccccccac ggacttggga tgagtttctt gccacctgtc aaacacttaa acaaaagggt
     1921 ctggaagctc cactggctct cggggaaaat tggacacaac aacacctctg ggagagtgtc
     1981 gccttggctg tattggggcc tgatgattgg aataatctgt ggaatgggaa gctgaagttc
     2041 acagacccta aagcagttag ggcatgggaa gtctttggcc gcgttttgga ctgtgccaac
     2101 aaggatgcag caggcttgtc ttggcagcaa gctgtcgatc gggtagtaca ggggaaagcc
     2161 gcttttaacg taatgggcga ctgggcagca ggatatatga ctaccactct caagttgaag
     2221 cctggtacag atttcgcatg ggcaccaagt cccgggacac agggcgtctt tatgatggtt
     2281 tccgactctt tcggacttcc caaaggagcc aaaaatcgac aaaacgctat caactggctt
     2341 cgattggtcg gctctaaaga aggtcaagat acattcaacc cactgaaagg gtccatcgct
     2401 gctcggctcg atagcgaccc aagcaagtat ccgtgtaacg tctatatcac cgccgacaag
     2461 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acgtggagga cggcagcgtg
     2521 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     2581 gacaaccact acctggtgac ccagtccgtg ctgagcaaag accccaacga gaagcgcgat
     2641 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     2701 tacaagggcg gtaccggagg gagcatgagc aagggcgagg agctgttcac cggggtggtg
     2761 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gcgcggcgag
     2821 ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag
     2881 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     2941 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     3001 gtccaggagc gcaccatcag cttcaaggac gacggcacct acaagacccg cgccgaggtg
     3061 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     3121 gacggcaaca tcctggggca caagctggag tacaacttta acaaccccaa cgcatacgga
     3181 cagagtgcaa tgcgagattg gcggagtaat cgcatcgttg gctccctggt agcaggagca
     3241 gtggcccctg aaagctccat gtcacaattc ggaacagtaa tggaaatttt cctgcaaaca
     3301 cgcaacccac aagcagcagc aaatgcagca caagctattg ctgaccaagt tggcctgggt
     3361 cgccttggtc agctgcagtc taccggctcc ggaaatgctg tgggccagga cacgcaggag
     3421 cgtatggtgt ctaagggcga agagctgatc aaggaaaata tgcgtatgaa ggtggtcatg
     3481 gaaggttcgg tcaacggcca ccaattcaaa tgcacaggtg aaggagaagg caggccgtac
     3541 gagggagtgc aaaccatgag gatcaaagtc atcgagggag gacccctgcc atttgccttt
     3601 gacattcttg ccacgtcgtt catgtatggc agccgtactt ttatcaagta cccggcagac
     3661 attcctgatt tctttaaaca gtcctttcct gagggtttta cttgggaaag agttacgaga
     3721 tacgaagatg gtggagtcgt caccgtcacg caggacacca gccttgagga tggcgaactc
     3781 gtttacaacg tcaaagtcag aggggtaaac tttccctcca atggtcccgt gatgcagaag
     3841 aagaccaagg gttgggagcc taatacagag atgatgtatc cagcagatgg tggtctgagg
     3901 ggatacactg atattgcact gaaagttgat ggtggtggcc atctgcattg caatttcgta
     3961 acaacttaca ggtcaaaaaa gaccgtcggg aacatcaaga tgcccggtgt ccatgccgtt
     4021 gatcaccgcc tggaaaggat agaggaaagt gacaatgaaa cgtacgtagt acaacgcgaa
     4081 gtcgcagttg ccaagtactc caatcttggt ggtggtatgg atgaacttta taagtcccat
     4141 caccatcacc accattaaga attcgaagct tttcgaatta gcttatcata atcaacctct
     4201 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     4261 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     4321 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     4381 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     4441 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     4501 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     4561 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     4621 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     4681 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     4741 tcagacgagt cggatctccc tttgggccgc ctccccgcat cggaggccgc ttcgagcaga
     4801 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     4861 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     4921 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga
     4981 ggttttttaa agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag
     5041 agcatggcta cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
     5101 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     5161 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     5221 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     5281 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5341 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5401 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5461 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5521 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5581 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5641 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5701 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5761 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5821 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5881 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     5941 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     6001 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     6061 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     6121 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     6181 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     6241 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     6301 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     6361 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     6421 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     6481 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     6541 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     6601 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     6661 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     6721 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     6781 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     6841 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     6901 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     6961 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     7021 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     7081 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     7141 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcc
//
