LOCUS       sequence-487065-        7362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic expression of green glucose sensor (high affinity) with 
            non-responsive HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7362)
  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 7362)
  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.H348H.HaloTag
FEATURES             Location/Qualifiers
     source          1..7362
                     /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             4312..4329
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    4364..4952
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4417..4437)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4835..4846)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4980..5101
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4991..5010)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5045..5064
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   5263..5392
                     /label=AAV2 ITR
     rep_origin      complement(5629..6217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5709..5728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6388..7179,7180..7248))
                     /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(7249..7353)
                     /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 acacggagca
     3241 gtggcccctg aaagctccat gtcacaattc ggaacagtaa tggaaatttt cctgcaaaca
     3301 cgcaacccac aagcagcagc aaatgcagca caagctattg ctgaccaagt tggcctgggt
     3361 cgccttggtc agctgcaggg cggaggctcg ggcggaggtg ggtcgggtgg cggcggatca
     3421 atggccgaga tcggcaccgg cttccccttc gacccccact acgtggaggt gctgggcgag
     3481 cgcatgcact acgttgacgt gggcccccgc gacggcaccc ccgtgctgtt cctgcacggc
     3541 aaccccacca gcagctacgt gtggcgcaac atcatccccc acgtggcccc cacccaccgc
     3601 actatcgccc ccgacctgat cggcatgggc aagagcgaca agcccgacct gggctacttc
     3661 ttcgacgacc acgtgcgctt catggacgcc ttcatcgagg ccctgggcct ggaggaggtg
     3721 gtgctggtga tccacgactg gggcagcgcc ctgggcttcc actgggccaa gcgcaacccc
     3781 gagcgcgtga agggcatcgc cttcatggag ttcatccgcc ccatccccac ctgggacgag
     3841 tggcccgagt tcgcccgcga gaccttccag gccttccgca ccaccgacgt gggccgcaag
     3901 ctgatcatcg accagaacgt gttcatcgag ggcaccctgc ccatgggcgt ggtgcgcccc
     3961 ctgaccgagg tggagatgga ccactaccgc gagcccttcc tgaaccccgt ggaccgcgag
     4021 cccctgtggc gcttccccaa cgagctgccc atcgccggcg agcccgccaa catcgtggcc
     4081 ctggtggagg agtacatgga ctggctgcac cagagccccg tgcccaagct gctgttctgg
     4141 ggcacccccg gcgtgctgat cccccccgcc gaggccgccc gcctggccaa gagcctgccc
     4201 aacgttaagg ccgtggacat cggccccggc ctgaacctgc tccaggagga caaccccgac
     4261 ctgatcggca gcgagatcgc ccgctggctg agcaccctcg agatcagcgg ccatcaccat
     4321 caccaccatt aagaattcga agcttttcga attagcttat cataatcaac ctctggatta
     4381 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     4441 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     4501 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     4561 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     4621 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     4681 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4741 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     4801 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     4861 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     4921 gagtcggatc tccctttggg ccgcctcccc gcatcggagg ccgcttcgag cagacatgat
     4981 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     5041 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     5101 taacaacaac aattgcattc attttatgtt tcaggttcag ggggagatgt gggaggtttt
     5161 ttaaagcaag taaaacctct acaaatgtgg taaaatcgat aaggatcttc ctagagcatg
     5221 gctacgtaga taagtagcat ggcgggttaa tcattaacta caaggaaccc ctagtgatgg
     5281 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     5341 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcatta
     5401 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5461 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6061 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7321 tatttagaaa aataaacaaa taggggttcc gcgcacattt cc
//
