LOCUS       sequence-487023-        7948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ER targeted 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 7948)
  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 7948)
  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.(ER).iGlucoSnFR2.HaloTag
FEATURES             Location/Qualifiers
     source          1..7948
                     /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)."
     primer_bind     2528..2549
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2652..2673)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(4224..4257)
                     /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(4320..4353)
                     /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(4391..4410)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(4472..4494)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(4773..5050)
                     /label=chicken beta-actin promoter
     repeat_region   5361..5490
                     /label=AAV2 ITR
     rep_origin      5562..6017
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5649..5668)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5859..5880
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6043..6147
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6148..6216,6217..7008)
                     /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(6366..6385)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7179..7767
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7668..7687
                     /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 accatgctgc tatccgtgcc gctgctgctc ggcctcctcg
     1321 gcctggccat cgccagatct aagctggaga ttttctcttg gtgggctggg gatgaaggcc
     1381 ctgccctgga ggcactcatc agactgtata aacagaagta tcctggagta gaggtgataa
     1441 acgccactgt caccggaggt gccggtgtaa atgctcgcgc tgtccttaag acaaggatgc
     1501 ttggtggtga tcctcctgat acatttcaag ttgcagcagg gatggagttg ataggcacat
     1561 gggtggtggc taacagaatg gaggatctca gcgccttgtt tcgacaagag ggttggcttc
     1621 aagccttccc caagggcctt attgatctga ttagttataa aggtggtata tggagtgtgc
     1681 ccgttaatat tcacaggtca aacgtaatgt ggtatttgcc tgcaaagcta aaagagtggg
     1741 gagttaaccc cccacggact tgggatgagt ttcttgccac ctgtcaaaca cttaaacaaa
     1801 agggtctgga agctccactg gctctcgggg aaaattggac acaacaacac ctctgggaga
     1861 gtgtcgcctt ggctgtattg gggcctgatg attggaataa tctgtggaat gggaagctga
     1921 agttcacaga ccctaaagca gttagggcat gggaagtctt tggccgcgtt ttggactgtg
     1981 ccaacaagga tgcagcaggc ttgtcttggc agcaagctgt cgatcgggta gtacagggga
     2041 aagccgcttt taacgtaatg ggcgactggg cagcaggata tatgactacc actctcaagt
     2101 tgaagcctgg tacagatttc gcatgggcac caagtcccgg gacacagggc gtctttatga
     2161 tggtttccga ctctttcgga cttcccaaag gagccaaaaa tcgacaaaac gctatcaact
     2221 ggcttcgatt ggtcggctct aaagaaggtc aagatacatt caacccactg aaagggtcca
     2281 tcgctgctcg gctcgatagc gacccaagca agtatccgtg taacgtctat atcaccgccg
     2341 acaagcagaa gaacggcatc aaggcgaact tcaagatccg ccacaacgtg gaggacggca
     2401 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2461 tgcccgacaa ccactacctg gtgacccagt ccgtgctgag caaagacccc aacgagaagc
     2521 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2581 agctgtacaa gggcggtacc ggagggagca tgagcaaggg cgaggagctg ttcaccgggg
     2641 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgcgcg
     2701 gcgagggcga gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg
     2761 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2821 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2881 gctacgtcca ggagcgcacc atcagcttca aggacgacgg cacctacaag acccgcgccg
     2941 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3001 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctttaacaac cccaacgcat
     3061 acggacagag tgcaatgcga gattggcgga gtaatcgcat cgttggctcc ctggtagcag
     3121 gagcagtggc ccctgaaagc tccatgtcac aattcggaac agtaatggaa attttcctgc
     3181 aaacacgcaa cccacaagca gcagcaaatg cagcacaagc tattgctgac caagttggcc
     3241 tgggtcgcct tggtcagctg cagggcggag gctcgggcgg aggtgggtcg ggtggcggcg
     3301 gatcaatggc cgagatcggc accggcttcc ccttcgaccc ccactacgtg gaggtgctgg
     3361 gcgagcgcat gcactacgtt gacgtgggcc cccgcgacgg cacccccgtg ctgttcctgc
     3421 acggcaaccc caccagcagc tacgtgtggc gcaacatcat cccccacgtg gcccccaccc
     3481 accgcactat cgcccccgac ctgatcggca tgggcaagag cgacaagccc gacctgggct
     3541 acttcttcga cgaccacgtg cgcttcatgg acgccttcat cgaggccctg ggcctggagg
     3601 aggtggtgct ggtgatccac gactggggca gcgccctggg cttccactgg gccaagcgca
     3661 accccgagcg cgtgaagggc atcgccttca tggagttcat ccgccccatc cccacctggg
     3721 acgagtggcc cgagttcgcc cgcgagacct tccaggcctt ccgcaccacc gacgtgggcc
     3781 gcaagctgat catcgaccag aacgtgttca tcgagggcac cctgcccatg ggcgtggtgc
     3841 gccccctgac cgaggtggag atggaccact accgcgagcc cttcctgaac cccgtggacc
     3901 gcgagcccct gtggcgcttc cccaacgagc tgcccatcgc cggcgagccc gccaacatcg
     3961 tggccctggt ggaggagtac atggactggc tgcaccagag ccccgtgccc aagctgctgt
     4021 tctggggcac ccccggcgtg ctgatccccc ccgccgaggc cgcccgcctg gccaagagcc
     4081 tgcccaacgt taaggccgtg gacatcggcc ccggcctgaa cctgctccag gaggacaacc
     4141 ccgacctgat cggcagcgag atcgcccgct ggctgagcac cctcgagatc agcggcaaag
     4201 atgagttgta gaagcttgct agtataactt cgtatagcat acattatacg aagttatggg
     4261 tcgatggtga tgcttggcaa ttcgggtcga tggtgaagca ttggtcttcc tattttgata
     4321 taacttcgta taggatactt tatacgaagt tatggggatg atccaattct ttgccaaaat
     4381 gatgagacag cacaataacc agcacgttgc ccaggagctg taggaaaaag aagaaggcat
     4441 gaacatggtt agcagaggct ctagagccgc cggtcacacg ccagaagccg aaccccgccc
     4501 tgccccgtcc cccccgaagg cagccgtccc cccgcggaca gccccgagcc gctccccccg
     4561 cacaaagggc cctcccggag cccctcaagg ctttcacgca gccacagaaa agaaacaagc
     4621 cgtcattaaa ccaagcgcta attacagccc ggaggagaag ggccgtcccg cccgctcacc
     4681 tgtgggagta acgcggtcag tcagagccgg ggcgggcggc gcgaggcggc ggcggagcgg
     4741 ggcacggggc gaaggcagcg cgcagcgact cccgcccgcc gcgcgcttcg ctttttatag
     4801 ggccgccgcc gccgccgcct cgccataaaa ggaaactttc ggagcgcgcc gctctgattg
     4861 gctgccgccg cacctctccg cctcgccccg ccccgcccct cgccccgccc cgccccgcct
     4921 ggcgcgcgcc cccccccccc ccccgccccc atcgctgcac aaaataatta aaaaataaat
     4981 aaatacaaaa ttgggggtgg ggaggggggg gagatgggga gagtgaagca gaacgtgggg
     5041 ctcacctcga ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     5101 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     5161 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     5221 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     5281 gtcattattg acgtcaatgg tactcttcct agagcatggc tacgtagata agtagcatgg
     5341 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
     5401 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     5461 ggcggcctca gtgagcgagc gagcgcgcag ctggcgtaat agcgaagagg cccgcaccga
     5521 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc
     5581 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     5641 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     5701 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     5761 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     5821 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     5881 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     5941 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     6001 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     6061 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     6121 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     6181 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     6241 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     6301 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     6361 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     6421 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     6481 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     6541 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     6601 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     6661 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     6721 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     6781 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     6841 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     6901 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     6961 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     7021 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     7081 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     7141 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     7201 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     7261 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     7321 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     7381 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     7441 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     7501 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     7561 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     7621 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     7681 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     7741 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     7801 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     7861 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     7921 agcgagtcag tgagcgagga agcggaag
//
