LOCUS       sequence-487016-        7534 bp    DNA     circular SYN 23-SEP-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 7534)
  TITLE     iGlucoSnFR2: a genetically encoded fluorescent sensor for measuring 
            intracellular or extracellular glucose in vivo in mouse brain
REFERENCE   2  (bases 1 to 7534)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 23, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pAAV.hSynapFLEX.(cyto).iGlucoSnFR2.mRuby3
FEATURES             Location/Qualifiers
     source          1..7534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      69..524
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(156..175)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     366..387
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        550..654
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(655..723,724..1515)
                     /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(873..892)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1686..2274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2175..2194
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   2511..2640
                     /label=AAV2 ITR
     polyA_signal    2802..2923
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2839..2858)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2893..2912
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     misc_feature    3001..3589
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3107..3118
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3516..3536
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    3661..3694
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3765..3798
                     /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      3823..3832
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     5028..5049
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5152..5173)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(6535..6568)
                     /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(6631..6664)
                     /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(6675..6694)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     intron          6677..6809
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     repeat_region   7402..7531
                     /label=AAV2 ITR
ORIGIN
        1 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
       61 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
      121 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
      181 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      241 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
      301 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      361 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
      421 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
      481 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
      541 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      601 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      661 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      721 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      781 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      841 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      901 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
      961 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1021 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1081 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1141 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1201 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1261 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1321 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1381 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1441 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1501 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1561 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1621 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1681 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1741 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1801 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1861 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1921 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1981 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2041 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2101 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2161 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2221 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2281 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2341 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2401 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2461 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctgcgcgctc
     2521 gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg
     2581 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
     2641 tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct aggaagatcc
     2701 ttatcgattt taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacatctcc
     2761 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     2821 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2881 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgctcgaagc
     2941 ggccgcccgg gtcgactcta gatgcatgct cgagcggccg cccgggtcga cggtatcgat
     3001 gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga agacgcggaa
     3061 gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg agggccgaag
     3121 ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacacaggcg agcagccaag
     3181 gaaaggacga tgatttcccc gacaacacca cggaattgtc agtgcccaac agccgagccc
     3241 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata gggaggggga
     3301 aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca gtgggggttg
     3361 cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac aactcctcat
     3421 aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
     3481 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
     3541 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta tcgataagct
     3601 tctcgagtgg atcatctgca gaattcgccc ttttcagtgt ttcgaacctc ttcgagggac
     3661 ataacttcgt ataatgtatg ctatacgaag ttatgggtcg atggtgagat ctggactaga
     3721 gggtcgatgg tgatgcttgg atcaaaatag gaagaatccc aatgataact tcgtataaag
     3781 tatcctatac gaagttatac tagtggtacc accggtgata tcgccaccat gggcagatct
     3841 aagctggaga ttttctcttg gtgggctggg gatgaaggcc ctgccctgga ggcactcatc
     3901 agactgtata aacagaagta tcctggagta gaggtgataa acgccactgt caccggaggt
     3961 gccggtgtaa atgctcgcgc tgtccttaag acaaggatgc ttggtggtga tcctcctgat
     4021 acatttcaag ttgcagcagg gatggagttg ataggcacat gggtggtggc taacagaatg
     4081 gaggatctca gcgccttgtt tcgacaagag ggttggcttc aagccttccc caagggcctt
     4141 attgatctga ttagttataa aggtggtata tggagtgtgc ccgttaatat tcacaggtca
     4201 aacgtaatgt ggtatttgcc tgcaaagcta aaagagtggg gagttaaccc cccacggact
     4261 tgggatgagt ttcttgccac ctgtcaaaca cttaaacaaa agggtctgga agctccactg
     4321 gctctcgggg aaaattggac acaacaacac ctctgggaga gtgtcgcctt ggctgtattg
     4381 gggcctgatg attggaataa tctgtggaat gggaagctga agttcacaga ccctaaagca
     4441 gttagggcat gggaagtctt tggccgcgtt ttggactgtg ccaacaagga tgcagcaggc
     4501 ttgtcttggc agcaagctgt cgatcgggta gtacagggga aagccgcttt taacgtaatg
     4561 ggcgactggg cagcaggata tatgactacc actctcaagt tgaagcctgg tacagatttc
     4621 gcatgggcac caagtcccgg gacacagggc gtctttatga tggtttccga ctctttcgga
     4681 cttcccaaag gagccaaaaa tcgacaaaac gctatcaact ggcttcgatt ggtcggctct
     4741 aaagaaggtc aagatacatt caacccactg aaagggtcca tcgctgctcg gctcgatagc
     4801 gacccaagca agtatccgtg taacgtctat atcaccgccg acaagcagaa gaacggcatc
     4861 aaggcgaact tcaagatccg ccacaacgtg gaggacggca gcgtgcagct cgccgaccac
     4921 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     4981 gtgacccagt ccgtgctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     5041 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggcggtacc
     5101 ggagggagca tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     5161 ctggacggcg acgtaaacgg ccacaagttc agcgtgcgcg gcgagggcga gggcgatgcc
     5221 accaacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     5281 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     5341 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     5401 atcagcttca aggacgacgg cacctacaag acccgcgccg aggtgaagtt cgagggcgac
     5461 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     5521 gggcacaagc tggagtacaa ctttaacaac cccaacgcat acggacagag tgcaatgcga
     5581 gattggcgga gtaatcgcat cgttggctcc ctggtagcag gagcagtggc ccctgaaagc
     5641 tccatgtcac aattcggaac agtaatggaa attttcctgc aaacacgcaa cccacaagca
     5701 gcagcaaatg cagcacaagc tattgctgac caagttggcc tgggtcgcct tggtcagctg
     5761 cagtctaccg gctccggaaa tgctgtgggc caggacacgc aggagcgtat ggtgtctaag
     5821 ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg tcatggaagg ttcggtcaac
     5881 ggccaccaat tcaaatgcac aggtgaagga gaaggcaggc cgtacgaggg ggtgcaaacc
     5941 atgaggatca aagtcatcga gggaggaccc ctgccatttg cctttgacat tcttgccacg
     6001 tcgttcatgt atggcagccg tacttttatc aagtacccgg cagacattcc tgatttcttt
     6061 aaacagtcct ttcctgaggg ttttacttgg gaaagagtta cgagatacga agatggtgga
     6121 gtcgtcaccg tcacgcagga caccagcctt gaggatggcg aactcgttta caacgtcaaa
     6181 gtcagagggg taaactttcc ctccaatggt cccgtgatgc agaagaagac caagggttgg
     6241 gagcctaata cagagatgat gtatccagca gatggtggtc tgaggggata cactgatatt
     6301 gcactgaaag ttgatggtgg tggccatctg cattgcaatt tcgtaacaac ttacaggtca
     6361 aaaaagaccg tcgggaacat caagatgccc ggtgtccatg ccgttgatca ccgcctggaa
     6421 aggatagagg aaagtgacaa tgaaacgtac gtagtacaac gcgaagtcgc agttgccaag
     6481 tactccaatc ttggtggtgg tatggatgaa ctttataagt aagctagcac tagtataact
     6541 tcgtatagca tacattatac gaagttatgt gtcgatggtg atgcttggca attcgggtcg
     6601 atggtgaagc attggtcttc ctattttgat ataacttcgt ataggatact ttatacgaag
     6661 ttatggggat catcacctgt ggagagaaag gcaaagtgga tgtcagtaag accaataggt
     6721 gcctatcaga aacgcaagag tcttctctgt ctcgacaagc ccagtttcta ttggtctcct
     6781 taaacctgtc ttgtaacctt gatacttacc tgatccaacg gctgcgactt ggggcagggg
     6841 gtcctagggg tggtctggcc aggagccgcg ggggcggact gcgcggtgcc caggagcaca
     6901 gctgcgctct caggcacgac acgactcctc cgctgcccac cgcagactga ggcagcgctg
     6961 agtcgccggc gccgcagcgc agatggtcgc gcccgtgccc ccctatctcg cgcctcgcgt
     7021 ggtgcggtcc ggctgggccg gcggcggcgc ggacgcgacc aaggtggccg ggaaggggag
     7081 tttgcggggg accggcgagt gacgtcagcg cgccttcagt gctgaggcgg cggtggcgcg
     7141 cgccgccagg cgggggcgaa ggcactgtcc gcggtgctga agctggcagt gcgcacgcgc
     7201 ctcgccgcat cctgtttccc ctccccctct ctgatagggg atgcgcaatt tggggaatgg
     7261 gggttgggtg cttgtccagt gggtcggggt cggtcgtcaa tttaaatact gaaaagggcg
     7321 aattccagca cactggagct cggatcttcc tagagcatgg ctacgtagat aagtagcatg
     7381 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     7441 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     7501 gggcggcctc agtgagcgag cgagcgcgca gctg
//
