LOCUS       sequence-487028-        6404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic expression of green glucose sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6404)
  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 6404)
  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
FEATURES             Location/Qualifiers
     source          1..6404
                     /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"
     misc_feature    3406..3994
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3459..3479)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3877..3888)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4022..4143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4033..4052)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4087..4106
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4305..4434
                     /label=AAV2 ITR
     rep_origin      complement(4671..5259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4751..4770)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5430..6221,6222..6290))
                     /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(6291..6395)
                     /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 agctgcagta gaagcttttc gaattagctt atcataatca acctctggat
     3421 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     3481 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     3541 tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg
     3601 caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc
     3661 accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa
     3721 ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat
     3781 tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc
     3841 tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt
     3901 ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag
     3961 acgagtcgga tctccctttg ggccgcctcc ccgcatcgga ggccgcttcg agcagacatg
     4021 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     4081 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4141 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     4201 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatct tcctagagca
     4261 tggctacgta gataagtagc atggcgggtt aatcattaac tacaaggaac ccctagtgat
     4321 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4381 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcat
     4441 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4501 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4561 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4621 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4681 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4741 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4801 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4861 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4921 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4981 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5041 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5101 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5161 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5221 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5281 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5341 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5401 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5461 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5521 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5581 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5641 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5701 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5761 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5821 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5881 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5941 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6001 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6061 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6121 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6181 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6241 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6301 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6361 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttcc
//
