LOCUS       sequence-487020-        6958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic expression of green glucose sensor with non-responsive 
            HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6958)
  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 6958)
  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.CAGFLEX.(cyto).iGlucoSnFR2
FEATURES             Location/Qualifiers
     source          1..6958
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   56..185
                     /label=AAV2 ITR
     polyA_signal    347..468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(384..403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     438..457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    511..1099
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             617..628
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1026..1046
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    1133..1166
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    1237..1270
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     regulatory      1278..1287
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2483..2504
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2607..2628)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(3234..3267)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(3330..3363)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(3401..3420)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3482..3504)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3783..4060)
                     /label=chicken beta-actin promoter
     repeat_region   4371..4500
                     /label=AAV2 ITR
     rep_origin      4572..5027
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4659..4678)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4869..4890
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5053..5157
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5158..5226,5227..6018)
                     /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     complement(5376..5395)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6189..6777
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6678..6697
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg cgtcgggcga cctttggtcg
      121 cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca tcactagggg
      181 ttccttgtag ttaatgatta acccgccatg ctacttatct acgtagccat gctctaggaa
      241 gatccttatc gattttacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      301 tctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgctc
      481 gaagcggccg cccgggtcga cggtatcgat gcggggaggc ggcccaaagg gagatccgac
      541 tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc aggccgcggg
      601 aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc ccgcgcagaa
      661 tccaggtggc aacacaggcg agcagccaag gaaaggacga tgatttcccc gacaacacca
      721 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      781 tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg agctgacagg
      841 tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac accacgccac
      901 gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat ttatacaagg
      961 aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt aaagcagcgt
     1021 atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt tcacaaattt
     1081 tgtaatccag aggttgatta tcgataagct aattcgaacc tcttcgaggg acataacttc
     1141 gtataatgta tgctatacga agttatgggt cgatggtgag atctggacta gagggtcgat
     1201 ggtgatgctt ggatcaaaat aggaagaatc ccaatgataa cttcgtataa agtatcctat
     1261 acgaagttat ggatcccgcc accatgggca gatctaagct ggagattttc tcttggtggg
     1321 ctggggatga aggccctgcc ctggaggcac tcatcagact gtataaacag aagtatcctg
     1381 gagtagaggt gataaacgcc actgtcaccg gaggtgccgg tgtaaatgct cgcgctgtcc
     1441 ttaagacaag gatgcttggt ggtgatcctc ctgatacatt tcaagttgca gcagggatgg
     1501 agttgatagg cacatgggtg gtggctaaca gaatggagga tctcagcgcc ttgtttcgac
     1561 aagagggttg gcttcaagcc ttccccaagg gccttattga tctgattagt tataaaggtg
     1621 gtatatggag tgtgcccgtt aatattcaca ggtcaaacgt aatgtggtat ttgcctgcaa
     1681 agctaaaaga gtggggagtt aaccccccac ggacttggga tgagtttctt gccacctgtc
     1741 aaacacttaa acaaaagggt ctggaagctc cactggctct cggggaaaat tggacacaac
     1801 aacacctctg ggagagtgtc gccttggctg tattggggcc tgatgattgg aataatctgt
     1861 ggaatgggaa gctgaagttc acagacccta aagcagttag ggcatgggaa gtctttggcc
     1921 gcgttttgga ctgtgccaac aaggatgcag caggcttgtc ttggcagcaa gctgtcgatc
     1981 gggtagtaca ggggaaagcc gcttttaacg taatgggcga ctgggcagca ggatatatga
     2041 ctaccactct caagttgaag cctggtacag atttcgcatg ggcaccaagt cccgggacac
     2101 agggcgtctt tatgatggtt tccgactctt tcggacttcc caaaggagcc aaaaatcgac
     2161 aaaacgctat caactggctt cgattggtcg gctctaaaga aggtcaagat acattcaacc
     2221 cactgaaagg gtccatcgct gctcggctcg atagcgaccc aagcaagtat ccgtgtaacg
     2281 tctatatcac cgccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca
     2341 acgtggagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     2401 acggccccgt gctgctgccc gacaaccact acctggtgac ccagtccgtg ctgagcaaag
     2461 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     2521 ctctcggcat ggacgagctg tacaagggcg gtaccggagg gagcatgagc aagggcgagg
     2581 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2641 agttcagcgt gcgcggcgag ggcgagggcg atgccaccaa cggcaagctg accctgaagt
     2701 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2761 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2821 ccgccatgcc cgaaggctac gtccaggagc gcaccatcag cttcaaggac gacggcacct
     2881 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2941 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacttta
     3001 acaaccccaa cgcatacgga cagagtgcaa tgcgagattg gcggagtaat cgcatcgttg
     3061 gctccctggt agcaggagca gtggcccctg aaagctccat gtcacaattc ggaacagtaa
     3121 tggaaatttt cctgcaaaca cgcaacccac aagcagcagc aaatgcagca caagctattg
     3181 ctgaccaagt tggcctgggt cgccttggtc agctgcagta gaagcttgct agtataactt
     3241 cgtatagcat acattatacg aagttatggg tcgatggtga tgcttggcaa ttcgggtcga
     3301 tggtgaagca ttggtcttcc tattttgata taacttcgta taggatactt tatacgaagt
     3361 tatggggatg atccaattct ttgccaaaat gatgagacag cacaataacc agcacgttgc
     3421 ccaggagctg taggaaaaag aagaaggcat gaacatggtt agcagaggct ctagagccgc
     3481 cggtcacacg ccagaagccg aaccccgccc tgccccgtcc cccccgaagg cagccgtccc
     3541 cccgcggaca gccccgagcc gctccccccg cacaaagggc cctcccggag cccctcaagg
     3601 ctttcacgca gccacagaaa agaaacaagc cgtcattaaa ccaagcgcta attacagccc
     3661 ggaggagaag ggccgtcccg cccgctcacc tgtgggagta acgcggtcag tcagagccgg
     3721 ggcgggcggc gcgaggcggc ggcggagcgg ggcacggggc gaaggcagcg cgcagcgact
     3781 cccgcccgcc gcgcgcttcg ctttttatag ggccgccgcc gccgccgcct cgccataaaa
     3841 ggaaactttc ggagcgcgcc gctctgattg gctgccgccg cacctctccg cctcgccccg
     3901 ccccgcccct cgccccgccc cgccccgcct ggcgcgcgcc cccccccccc ccccgccccc
     3961 atcgctgcac aaaataatta aaaaataaat aaatacaaaa ttgggggtgg ggaggggggg
     4021 gagatgggga gagtgaagca gaacgtgggg ctcacctcga ccatggtaat agcgatgact
     4081 aatacgtaga tgtactgcca agtaggaaag tcccataagg tcatgtactg ggcataatgc
     4141 caggcgggcc atttaccgtc attgacgtca atagggggcg tacttggcat atgatacact
     4201 tgatgtactg ccaagtgggc agtttaccgt aaatactcca cccattgacg tcaatggaaa
     4261 gtccctattg gcgttactat gggaacatac gtcattattg acgtcaatgg tactcttcct
     4321 agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct
     4381 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
     4441 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
     4501 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     4561 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4621 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4681 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4741 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     4801 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4861 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4921 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4981 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     5041 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5101 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5161 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5221 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5281 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5341 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5401 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5461 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5521 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5581 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5641 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5701 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5761 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5821 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5881 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5941 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6001 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6061 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6121 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6181 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6241 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6301 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6361 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6421 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6481 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6541 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6601 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6661 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6721 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6781 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6841 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6901 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaag
//
