LOCUS       sequence-487026-        7647 bp    DNA     circular SYN 23-SEP-2025
DEFINITION  Plasma membrane targeted 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 7647)
  TITLE     iGlucoSnFR2: a genetically encoded fluorescent sensor for measuring 
            intracellular or extracellular glucose in vivo in mouse brain
REFERENCE   2  (bases 1 to 7647)
  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.CAG.(mem).iGlucoSnFR2.mRuby3
FEATURES             Location/Qualifiers
     source          1..7647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   61..190
                     /label=AAV2 ITR
     rep_origin      complement(427..1015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(507..526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1186..1977,1978..2046))
                     /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     1809..1828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2047..2151)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2177..2632)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2314..2335)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2526..2545
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2704..2833
                     /label=AAV2 ITR
     promoter        3146..3423
                     /label=chicken beta-actin promoter
     primer_bind     3702..3724
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3786..3805
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3839..3848
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     3845..3907
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     primer_bind     5101..5122
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5225..5246)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             5843..5872
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5876..6022
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     misc_feature    6794..7382
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6847..6867)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7265..7276)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    7425..7546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7436..7455)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7490..7509
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 ggatcttcct agagcatggc tacgtagata agtagcatgg cgggttaatc attaactaca
       61 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
      121 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
      181 gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
      241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
      301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
      361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
      421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
      481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
      541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
      601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
      661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
      721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
      781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
      841 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
      901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
      961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     1261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     1321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     1381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     1441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     1501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     1561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     1621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     1681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     1741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     1801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     1861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     1921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     1981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2161 ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2221 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2281 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2341 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2401 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2461 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2521 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     2581 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg
     2641 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     2701 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc
     2761 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     2821 actaggggtt ccttgtagtt aatgattaac ccgccatgct acttatctac gtagccatgc
     2881 tctaggaaga tcgtaccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     2941 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     3001 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     3061 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     3121 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
     3181 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
     3241 gcgatggggg cggggggggg gggggggcgc gcgccaggcg gggcggggcg gggcgagggg
     3301 cggggcgggg cgaggcggag aggtgcggcg gcagccaatc agagcggcgc gctccgaaag
     3361 tttcctttta tggcgaggcg gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg
     3421 gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc
     3481 ccgccccggc tctgactgac cgcgttactc ccacaggtga gcgggcggga cggcccttct
     3541 cctccgggct gtaattagcg cttggtttaa tgacggcttg tttcttttct gtggctgcgt
     3601 gaaagccttg aggggctccg ggagggccct ttgtgcgggg ggagcggctc ggggctgtcc
     3661 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     3721 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     3781 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tggatcccgc
     3841 caccatggag acagacacac tcctgctatg ggtactgctg ctctgggttc caggttccac
     3901 tggtgacaga tctaagctgg agattttctc ttggtgggct ggggatgaag gccctgccct
     3961 ggaggcactc atcagactgt ataaacagaa gtatcctgga gtagaggtga taaacgccac
     4021 tgtcaccgga ggtgccggtg taaatgctcg cgctgtcctt aagacaagga tgcttggtgg
     4081 tgatcctcct gatacatttc aagttgcagc agggatggag ttgataggca catgggtggt
     4141 ggctaacaga atggaggatc tcagcgcctt gtttcgacaa gagggttggc ttcaagcctt
     4201 ccccaagggc cttattgatc tgattagtta taaaggtggt atatggagtg tgcccgttaa
     4261 tattcacagg tcaaacgtaa tgtggtattt gcctgcaaag ctaaaagagt ggggagttaa
     4321 ccccccacgg acttgggatg agtttcttgc cacctgtcaa acacttaaac aaaagggtct
     4381 ggaagctcca ctggctctcg gggaaaattg gacacaacaa cacctctggg agagtgtcgc
     4441 cttggctgta ttggggcctg atgattggaa taatctgtgg aatgggaagc tgaagttcac
     4501 agaccctaaa gcagttaggg catgggaagt ctttggccgc gttttggact gtgccaacaa
     4561 ggatgcagca ggcttgtctt ggcagcaagc tgtcgatcgg gtagtacagg ggaaagccgc
     4621 ttttaacgta atgggcgact gggcagcagg atatatgact accactctca agttgaagcc
     4681 tggtacagat ttcgcatggg caccaagtcc cgggacacag ggcgtcttta tgatggtttc
     4741 cgactctttc ggacttccca aaggagccaa aaatcgacaa aacgctatca actggcttcg
     4801 attggtcggc tctaaagaag gtcaagatac attcaaccca ctgaaagggt ccatcgctgc
     4861 tcggctcgat agcgacccaa gcaagtatcc gtgtaacgtc tatatcaccg ccgacaagca
     4921 gaagaacggc atcaaggcga acttcaagat ccgccacaac gtggaggacg gcagcgtgca
     4981 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     5041 caaccactac ctggtgaccc agtccgtgct gagcaaagac cccaacgaga agcgcgatca
     5101 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     5161 caagggcggt accggaggaa gcatgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     5221 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgc gcggcgaggg
     5281 cgagggcgat gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     5341 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     5401 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     5461 ccaggagcgc accatcagct tcaaggacga cggcacctac aagacccgcg ccgaggtgaa
     5521 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     5581 cggcaacatc ctggggcaca agctggagta caactttaac aaccccaacg catacggaca
     5641 gagtgcaatg cgagattggc ggagtaatcg catcgttggc tccctggtag caggagcagt
     5701 ggcccctgaa agctccatgt cacaattcgg aacagtaatg gaaattttcc tgcaaacacg
     5761 caacccacaa gcagcagcaa atgcagcaca agctattgct gaccaagttg gcctgggtcg
     5821 ccttggtcag ctgcaggtcg acgaacaaaa actcatctca gaagaggatc tgaatgctgt
     5881 gggccaggac acgcaggagg tcatcgtggt gccacactcc ttgcccttta aggtggtggt
     5941 gatctcagcc atcctggccc tggtggtgct caccatcatc tcccttatca tcctcatcat
     6001 gctttggcag aagaagccac gttctagagg ctccggaaat gctgtgggcc aggacacgca
     6061 ggagcgtatg gtgtctaagg gcgaagagct gatcaaggaa aatatgcgta tgaaggtggt
     6121 catggaaggt tcggtcaacg gccaccaatt caaatgcaca ggtgaaggag aaggcaggcc
     6181 gtacgaggga gtgcaaacca tgaggatcaa agtcatcgag ggaggacccc tgccatttgc
     6241 ctttgacatt cttgccacgt cgttcatgta tggcagccgt acttttatca agtacccggc
     6301 agacattcct gatttcttta aacagtcctt tcctgagggt tttacttggg aaagagttac
     6361 gagatacgaa gatggtggag tcgtcaccgt cacgcaggac accagccttg aggatggcga
     6421 actcgtttac aacgtcaaag tcagaggggt aaactttccc tccaatggtc ccgtgatgca
     6481 gaagaagacc aagggttggg agcctaatac agagatgatg tatccagcag atggtggtct
     6541 gaggggatac actgatattg cactgaaagt tgatggtggt ggccatctgc attgcaattt
     6601 cgtaacaact tacaggtcaa aaaagaccgt cgggaacatc aagatgcccg gtgtccatgc
     6661 cgttgatcac cgcctggaaa ggatagagga aagtgacaat gaaacgtacg tagtacaacg
     6721 cgaagtcgca gttgccaagt actccaatct tggtggtggt atggatgaac tttataagta
     6781 gaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct
     6841 taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc
     6901 tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct
     6961 ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga
     7021 cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc
     7081 tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac
     7141 aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt
     7201 tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt
     7261 cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc
     7321 tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc
     7381 gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat acattgatga
     7441 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     7501 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     7561 cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa
     7621 cctctacaaa tgtggtaaaa tcgataa
//
