LOCUS       sequence-487009-        7605 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic expression of green glucose sensor (high affinity) with 
            non-responsive HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7605)
  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 7605)
  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.CAG.(cyto).iGlucoSnFR2.H348H.HaloTag
FEATURES             Location/Qualifiers
     source          1..7605
                     /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..3422
                     /label=chicken beta-actin promoter
     primer_bind     3701..3723
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3785..3804
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      3838..3847
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     5043..5064
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5167..5188)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             6712..6729
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    6752..7340
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6805..6825)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7223..7234)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    7383..7504
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7394..7413)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7448..7467
                     /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 ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc
     3301 ggggcggggc gaggcggaga ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt
     3361 ttccttttat ggcgaggcgg cggcggcggc ggccctataa aaagcgaagc gcgcggcggg
     3421 cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc
     3481 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
     3541 ctccgggctg taattagcgc ttggtttaat gacggcttgt ttcttttctg tggctgcgtg
     3601 aaagccttga ggggctccgg gagggccctt tgtgcggggg gagcggctcg gggctgtccg
     3661 cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg
     3721 accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt cctacagctc
     3781 ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt ggatcccgcc
     3841 accatgggca gatctaagct ggagattttc tcttggtggg ctggggatga aggccctgcc
     3901 ctggaggcac tcatcagact gtataaacag aagtatcctg gagtagaggt gataaacgcc
     3961 actgtcaccg gaggtgccgg tgtaaatgct cgcgctgtcc ttaagacaag gatgcttggt
     4021 ggtgatcctc ctgatacatt tcaagttgca gcagggatgg agttgatagg cacatgggtg
     4081 gtggctaaca gaatggagga tctcagcgcc ttgtttcgac aagagggttg gcttcaagcc
     4141 ttccccaagg gccttattga tctgattagt tataaaggtg gtatatggag tgtgcccgtt
     4201 aatattcaca ggtcaaacgt aatgtggtat ttgcctgcaa agctaaaaga gtggggagtt
     4261 aaccccccac ggacttggga tgagtttctt gccacctgtc aaacacttaa acaaaagggt
     4321 ctggaagctc cactggctct cggggaaaat tggacacaac aacacctctg ggagagtgtc
     4381 gccttggctg tattggggcc tgatgattgg aataatctgt ggaatgggaa gctgaagttc
     4441 acagacccta aagcagttag ggcatgggaa gtctttggcc gcgttttgga ctgtgccaac
     4501 aaggatgcag caggcttgtc ttggcagcaa gctgtcgatc gggtagtaca ggggaaagcc
     4561 gcttttaacg taatgggcga ctgggcagca ggatatatga ctaccactct caagttgaag
     4621 cctggtacag atttcgcatg ggcaccaagt cccgggacac agggcgtctt tatgatggtt
     4681 tccgactctt tcggacttcc caaaggagcc aaaaatcgac aaaacgctat caactggctt
     4741 cgattggtcg gctctaaaga aggtcaagat acattcaacc cactgaaagg gtccatcgct
     4801 gctcggctcg atagcgaccc aagcaagtat ccgtgtaacg tctatatcac cgccgacaag
     4861 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acgtggagga cggcagcgtg
     4921 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     4981 gacaaccact acctggtgac ccagtccgtg ctgagcaaag accccaacga gaagcgcgat
     5041 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     5101 tacaagggcg gtaccggagg gagcatgagc aagggcgagg agctgttcac cggggtggtg
     5161 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gcgcggcgag
     5221 ggcgagggcg atgccaccaa cggcaagctg accctgaagt tcatctgcac caccggcaag
     5281 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     5341 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     5401 gtccaggagc gcaccatcag cttcaaggac gacggcacct acaagacccg cgccgaggtg
     5461 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     5521 gacggcaaca tcctggggca caagctggag tacaacttta acaaccccaa cgcatacgga
     5581 cagagtgcaa tgcgagattg gcggagtaat cgcatcgttg gctccctggt acacggagca
     5641 gtggcccctg aaagctccat gtcacaattc ggaacagtaa tggaaatttt cctgcaaaca
     5701 cgcaacccac aagcagcagc aaatgcagca caagctattg ctgaccaagt tggcctgggt
     5761 cgccttggtc agctgcaggg cggaggctcg ggcggaggtg ggtcgggtgg cggcggatca
     5821 atggccgaga tcggcaccgg cttccccttc gacccccact acgtggaggt gctgggcgag
     5881 cgcatgcact acgttgacgt gggcccccgc gacggcaccc ccgtgctgtt cctgcacggc
     5941 aaccccacca gcagctacgt gtggcgcaac atcatccccc acgtggcccc cacccaccgc
     6001 actatcgccc ccgacctgat cggcatgggc aagagcgaca agcccgacct gggctacttc
     6061 ttcgacgacc acgtgcgctt catggacgcc ttcatcgagg ccctgggcct ggaggaggtg
     6121 gtgctggtga tccacgactg gggcagcgcc ctgggcttcc actgggccaa gcgcaacccc
     6181 gagcgcgtga agggcatcgc cttcatggag ttcatccgcc ccatccccac ctgggacgag
     6241 tggcccgagt tcgcccgcga gaccttccag gccttccgca ccaccgacgt gggccgcaag
     6301 ctgatcatcg accagaacgt gttcatcgag ggcaccctgc ccatgggcgt ggtgcgcccc
     6361 ctgaccgagg tggagatgga ccactaccgc gagcccttcc tgaaccccgt ggaccgcgag
     6421 cccctgtggc gcttccccaa cgagctgccc atcgccggcg agcccgccaa catcgtggcc
     6481 ctggtggagg agtacatgga ctggctgcac cagagccccg tgcccaagct gctgttctgg
     6541 ggcacccccg gcgtgctgat cccccccgcc gaggccgccc gcctggccaa gagcctgccc
     6601 aacgttaagg ccgtggacat cggccccggc ctgaacctgc tccaggagga caaccccgac
     6661 ctgatcggca gcgagatcgc ccgctggctg agcaccctcg agatcagcgg ccatcaccat
     6721 caccaccatt aagaattcga agcttatcga taatcaacct ctggattaca aaatttgtga
     6781 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     6841 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     6901 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     6961 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     7021 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     7081 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     7141 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     7201 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     7261 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     7321 cctttgggcc gcctccccgc atcgataccg tcgacccggg cggccgcttc gagcagacat
     7381 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     7441 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     7501 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt
     7561 tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataa
//
