LOCUS       sequence-487024-        7456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic expression of green glucose sensor with non-responsive 
            mIRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7456)
  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 7456)
  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.mIRFP670nano3
FEATURES             Location/Qualifiers
     source          1..7456
                     /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(3732..3765)
                     /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(3828..3861)
                     /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(3899..3918)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(3980..4002)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(4281..4558)
                     /label=chicken beta-actin promoter
     repeat_region   4869..4998
                     /label=AAV2 ITR
     rep_origin      5070..5525
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5157..5176)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5367..5388
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5551..5655
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5656..5724,5725..6516)
                     /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(5874..5893)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6687..7275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7176..7195
                     /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 agctgcagtc taccggctcc ggaggtggag
     3241 gtactggagg aggtgggtca ggtggaggtg gatcaatggc aaacctggac aagatgctga
     3301 acaccaccgt gaccgaggtg cgcaagttcc tgcaagcgga cagagtgtgc gtgttcaagt
     3361 tcgaggaaga ttactccggc accgtctcgc acgaagccgt ggacgacaga tggattagca
     3421 tcctgaagac ccaggtgcag gacagatact tcatggaaac cagaggcgag gaatacgtcc
     3481 acggcagata ccaggccatc gccgacatct acacagccaa tctggtcgag tgctacagag
     3541 acctgctgat cgagtttcag gtgcgggcca ttctggctgt ccccatcctg caaggcaaga
     3601 agctgtgggg cctgctggtg gcccaccaac tggccggccc tcgggagtgg cagacctggg
     3661 aaatcgactt cctgaaacag caagccgtgg tgatgggcat cgccatccag cagagctaga
     3721 agcttgctag tataacttcg tatagcatac attatacgaa gttatgggtc gatggtgatg
     3781 cttggcaatt cgggtcgatg gtgaagcatt ggtcttccta ttttgatata acttcgtata
     3841 ggatacttta tacgaagtta tggggatgat ccaattcttt gccaaaatga tgagacagca
     3901 caataaccag cacgttgccc aggagctgta ggaaaaagaa gaaggcatga acatggttag
     3961 cagaggctct agagccgccg gtcacacgcc agaagccgaa ccccgccctg ccccgtcccc
     4021 cccgaaggca gccgtccccc cgcggacagc cccgagccgc tccccccgca caaagggccc
     4081 tcccggagcc cctcaaggct ttcacgcagc cacagaaaag aaacaagccg tcattaaacc
     4141 aagcgctaat tacagcccgg aggagaaggg ccgtcccgcc cgctcacctg tgggagtaac
     4201 gcggtcagtc agagccgggg cgggcggcgc gaggcggcgg cggagcgggg cacggggcga
     4261 aggcagcgcg cagcgactcc cgcccgccgc gcgcttcgct ttttataggg ccgccgccgc
     4321 cgccgcctcg ccataaaagg aaactttcgg agcgcgccgc tctgattggc tgccgccgca
     4381 cctctccgcc tcgccccgcc ccgccccycg ccccgccccg ccccgcctgg cgcgcgcccc
     4441 cccccccccc ccgcccccat cgctgcacaa aataattaaa aaataaataa atacaaaatt
     4501 gggggtgggg agggggggga gatggggaga gtgaagcaga acgtggggct cacctcgacc
     4561 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     4621 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     4681 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     4741 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     4801 gtcaatggta ctcttcctag agcatggcta cgtagataag tagcatggcg ggttaatcat
     4861 taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4921 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4981 gagcgagcga gcgcgcagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
     5041 acagttgcgc agcctgaatg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
     5101 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     5161 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     5221 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     5281 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     5341 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     5401 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5461 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     5521 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5581 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5641 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5701 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5761 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5821 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5881 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5941 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6001 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6061 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6121 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6181 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6241 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6301 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6361 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6421 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6481 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6541 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6601 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6661 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6721 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6781 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6841 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6901 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6961 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7021 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7081 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7141 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7201 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7261 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7321 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7381 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     7441 agcgaggaag cggaag
//
