LOCUS       sequence-248792-        6745 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the iGlucoSnFR-TS biosensor for intracellular [glucose], 
            with a fluorescence-lifetime readout, in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6745)
  AUTHORS   Diaz-Garcia CM, Lahmann C, Martinez-Francois JR, Li B, Koveal D, 
            Nathwani N, Rahman M, Keller JP, Marvin JS, Looger LL, Yellen G
  TITLE     Quantitative in vivo imaging of neuronal glucose concentrations with
            a genetically encoded fluorescence lifetime sensor.
  JOURNAL   J Neurosci Res. 2019 May 20. doi: 10.1002/jnr.24433.
  PUBMED    31106909
REFERENCE   2  (bases 1 to 6745)
  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: "J Neurosci 
            Res. 2019 May 20. doi: 10.1002/jnr.24433."
COMMENT     Sequence Label: pAAV-CAG-iGlucoSnFR-TS
FEATURES             Location/Qualifiers
     source          1..6745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(182..770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(262..281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(941..1732,1733..1801))
                     /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     1564..1583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1802..1906)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1932..2387)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2069..2090)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2281..2300
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2459..2588
                     /label=AAV2 ITR
     promoter        2899..3176
                     /label=chicken beta-actin promoter
     primer_bind     3455..3477
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3539..3558
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3606..3623
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3627..3659
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(3663..3671,3672..3686)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     misc_feature    5647..6235
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5700..5720)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6118..6129)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    6278..6399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6289..6308)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6343..6362
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   6561..6690
                     /label=AAV2 ITR
ORIGIN
        1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
       61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
      121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
      181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
      841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
      901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
      961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
     2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2461 gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg
     2521 tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag
     2581 gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc catgctctag
     2641 gaagagtacc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2701 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2761 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     2821 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     2881 tcatcgctat taccatggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc
     2941 ccccctcccc acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg
     3001 gggcgggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg
     3061 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
     3121 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
     3181 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
     3241 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
     3301 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
     3361 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggctg tccgcggggg
     3421 gacggctgcc ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc
     3481 ggctctagag cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc
     3541 aacgtgctgg ttattgtgct gtctcatcat tttggcaaag aattggtacc accatgcggg
     3601 gttctcatca tcatcatcat catggtatgg ctagcatgac tggtggacag caaatgggtc
     3661 gggatctgta cgacgatgac gataaggacc gatggggatc caaactggaa atttttagct
     3721 ggtgggccgg cgatgaaggc ccggccctgg aagccctgat tcgcctgtat aaacagaaat
     3781 atccgggcgt ggaggtgatt aacgccaccg tgaccggcgg cgccggcgtg aacgcccgcg
     3841 ccgtgctgaa aacccgcatg ctgggcggcg atccgccgga tacctttcag gtggcagccg
     3901 gcatggaact gattggcacc tgggtggtgg ccaaccgcat ggaagacctg agcgccctgt
     3961 ttcgccagga aggctggctg caagcctttc cgaaaggcct gattgatctg attagctata
     4021 aaggcggcat ttggagcgtg ccggtgaaca ttcatcgcag caacgtgatg tggtatctgc
     4081 cggccaaact gaaagaatgg ggcgtgaacc cgccgcgcac ctgggatgaa tttctggcca
     4141 cctgccagac cctgaaacag aaaggcctgg aagccccgct ggccctgggc gaaaactgga
     4201 cccagcagca tctgtgggaa agcgtggccc tggctgtgct gggcccggat gattggaaca
     4261 acctgtggaa cggcaaactg aaatttaccg atccgaaagc cgtgcgcgcc tgggaagtgt
     4321 ttggccgcgt gctggattgc gccaacaaag atgccgccgg cctgagctgg cagcaggccg
     4381 tggatcgcgt ggtgcagggc aaagccgcct ttaacgtgat gggcgattgg gccgccggct
     4441 atatgaccac caccctgaaa ctgaaaccgg gcaccgattt tgcctgggcc ccgagcccgg
     4501 gcacccaggg cgtgtttatg atgctgagcg atagctttgg cctgccgaaa ggcgccaaaa
     4561 accgccagaa cgccattaac tggctgcgcc tggtgggcag caaagaaggc caggatacct
     4621 ttaacccgct gaaaggcagc attgccgccc gcctggattc tgatccgagt aaatatcccg
     4681 attctttcac tgctcacaac gtgtatatca tggctgacaa acagaagaac ggtatcaagg
     4741 ccaacttcaa gatccgccac aacatcgaag acggcggcgt tcagctggct gatcactacc
     4801 agcagaacac cccaatcggc gacggccctg ttctgctgcc tgacaaccac tacttaagca
     4861 tccagtctaa actgtccaaa gacccaaacg agaagcgcga ccacatggtt ctgctggaat
     4921 ttgttaccgc cgctggcatc acccacggca tggacgaact gtacaagggc ggcaccgggg
     4981 gatctatggt ttccaaaggc gaggaactgt tcactggtgt tgttcctatc ctggttgaac
     5041 tggacggcga cgttaacggc cacaaattca gcgttagcgg ggaaggcgaa ggagacgcta
     5101 cctacggcaa gctgactctg aagttcatct gcaccactgg caagctgcct gttccttggc
     5161 ctaccctggt tactaccttc tcctatggtg ttatggtttt cgccagatat cctgaccaca
     5221 tgaagcagca cgacttcttc aagagcgcca tgcctgaggg ctacgttcag gaacgcacca
     5281 tctttttcaa ggacgacggc aactataaga cacgcgctga ggttaagttc gagggcgaca
     5341 ctctggttaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     5401 gccataagct tgaatataac gagcccaacg cctatggtca gagtgcaatg agagattggc
     5461 gcagcaacag aattgtgggc agcctggtgg caggcgccgt ggccccggaa agctttatga
     5521 gccagtttgg caccgtgatg gaaatttttc tgcaaacccg caacccgcag gccgccgcca
     5581 acgccgccca ggccattgcc gatcaggtgg gcctgggccg cctgggccag taagactagt
     5641 atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     5701 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     5761 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     5821 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     5881 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     5941 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     6001 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg
     6061 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     6121 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     6181 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcatcga
     6241 taccgtcgac ccgggcggcc gcttcgagca gacatgataa gatacattga tgagtttgga
     6301 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     6361 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     6421 tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta aaacctctac
     6481 aaatgtggta aaatcgataa ggatcttcct agagcatggc tacgtagata agtagcatgg
     6541 cgggttaatc attaactaca aggaacccct agtgatggag ttggccactc cctctctgcg
     6601 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     6661 ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat gaatcggcca acgcgcgggg
     6721 agaggcggtt tgcgtattgg gcgct
//
