LOCUS       sequence-487000-        7337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cytosolic expression of green glucose sensor with non-responsive 
            mIRFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7337)
  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 7337)
  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.TBG.(cyto).iGlucoSnFR2.mIRFP670nano3
FEATURES             Location/Qualifiers
     source          1..7337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    49..70
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(85..90,91..108,109..115)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    123..139
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     128..150
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     147..163
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     147..163
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   201..330
                     /label=AAV2 ITR
     promoter        670..1079
                     /label=TBG promoter
                     /note="promoter from the human thyroxine-binding globulin 
                     gene (Yan et al., 2012)"
     intron          1139..1271
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1254..1273
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1283..1292
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2488..2509
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2612..2633)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3790..3810)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4208..4219)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4286..4303)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4292..4499
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4587..4716
                     /label=AAV2 ITR
     primer_bind     complement(4735..4752)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4735..4751)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4744..4766)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4893..5348
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4980..4999)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5190..5211
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5374..5478
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5479..5547,5548..6339)
                     /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(5697..5716)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6510..7098
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6999..7018
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
       61 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
      121 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccagatt
      181 taattaaggc cttaattagg ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc
      241 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
      301 gagtggccaa ctccatcact aggggttcct tgtagttaat gattaacccg ccatgctact
      361 tatctacgta gccatgctct aggaagatcg gaattcgccc ttaagctagc aggttaattt
      421 ttaaaaagca gtcaaaagtc caagtggccc ttggcagcat ttactctctc tgtttgctct
      481 ggttaataat ctcaggagca caaacattcc agatccaggt taatttttaa aaagcagtca
      541 aaagtccaag tggcccttgg cagcatttac tctctctgtt tgctctggtt aataatctca
      601 ggagcacaaa cattccagat ccggcgcgcc agggctggaa gctacctttg acatcatttc
      661 ctctgcgaat gcatgtataa tttctacaga acctattaga aaggatcacc cagcctctgc
      721 ttttgtacaa ctttccctta aaaaactgcc aattccactg ctgtttggcc caatagtgag
      781 aactttttcc tgctgcctct tggtgctttt gcctatggcc cctattctgc ctgctgaaga
      841 cactcttgcc agcatggact taaacccctc cagctctgac aatcctcttt ctcttttgtt
      901 ttacatgaag ggtctggcag ccaaagcaat cactcaaagt tcaaacctta tcattttttg
      961 ctttgttcct cttggccttg gttttgtaca tcagctttga aaataccatc ccagggttaa
     1021 tgctggggtt aatttataac taagagtgct ctagttttgc aatacaggac atgctataaa
     1081 aatggaaaga tgttgctttc tgagagactg cagaagttgg tcgtgaggca ctgggcaggt
     1141 aagtatcaag gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca
     1201 gagaagactc ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt
     1261 tctctccaca ggtgtccagg cggccgccat gggcagatct aagctggaga ttttctcttg
     1321 gtgggctggg gatgaaggcc ctgccctgga ggcactcatc agactgtata aacagaagta
     1381 tcctggagta gaggtgataa acgccactgt caccggaggt gccggtgtaa atgctcgcgc
     1441 tgtccttaag acaaggatgc ttggtggtga tcctcctgat acatttcaag ttgcagcagg
     1501 gatggagttg ataggcacat gggtggtggc taacagaatg gaggatctca gcgccttgtt
     1561 tcgacaagag ggttggcttc aagccttccc caagggcctt attgatctga ttagttataa
     1621 aggtggtata tggagtgtgc ccgttaatat tcacaggtca aacgtaatgt ggtatttgcc
     1681 tgcaaagcta aaagagtggg gagttaaccc cccacggact tgggatgagt ttcttgccac
     1741 ctgtcaaaca cttaaacaaa agggtctgga agctccactg gctctcgggg aaaattggac
     1801 acaacaacac ctctgggaga gtgtcgcctt ggctgtattg gggcctgatg attggaataa
     1861 tctgtggaat gggaagctga agttcacaga ccctaaagca gttagggcat gggaagtctt
     1921 tggccgcgtt ttggactgtg ccaacaagga tgcagcaggc ttgtcttggc agcaagctgt
     1981 cgatcgggta gtacagggga aagccgcttt taacgtaatg ggcgactggg cagcaggata
     2041 tatgactacc actctcaagt tgaagcctgg tacagatttc gcatgggcac caagtcccgg
     2101 gacacagggc gtctttatga tggtttccga ctctttcgga cttcccaaag gagccaaaaa
     2161 tcgacaaaac gctatcaact ggcttcgatt ggtcggctct aaagaaggtc aagatacatt
     2221 caacccactg aaagggtcca tcgctgctcg gctcgatagc gacccaagca agtatccgtg
     2281 taacgtctat atcaccgccg acaagcagaa gaacggcatc aaggcgaact tcaagatccg
     2341 ccacaacgtg gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2401 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg gtgacccagt ccgtgctgag
     2461 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2521 gatcactctc ggcatggacg agctgtacaa gggcggtacc ggagggagca tgagcaaggg
     2581 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2641 ccacaagttc agcgtgcgcg gcgagggcga gggcgatgcc accaacggca agctgaccct
     2701 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2761 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     2821 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcagcttca aggacgacgg
     2881 cacctacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2941 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3001 ctttaacaac cccaacgcat acggacagag tgcaatgcga gattggcgga gtaatcgcat
     3061 cgttggctcc ctggtagcag gagcagtggc ccctgaaagc tccatgtcac aattcggaac
     3121 agtaatggaa attttcctgc aaacacgcaa cccacaagca gcagcaaatg cagcacaagc
     3181 tattgctgac caagttggcc tgggtcgcct tggtcagctg cagtctaccg gctccggagg
     3241 tggaggtact ggaggaggtg ggtcaggtgg aggtggatca atggcaaacc tggacaagat
     3301 gctgaacacc accgtgaccg aggtgcgcaa gttcctgcaa gcggacagag tgtgcgtgtt
     3361 caagttcgag gaagattact ccggcaccgt ctcgcacgaa gccgtggacg acagatggat
     3421 tagcatcctg aagacccagg tgcaggacag atacttcatg gaaaccagag gcgaggaata
     3481 cgtccacggc agataccagg ccatcgccga catctacaca gccaatctgg tcgagtgcta
     3541 cagagacctg ctgatcgagt ttcaggtgcg ggccattctg gctgtcccca tcctgcaagg
     3601 caagaagctg tggggcctgc tggtggccca ccaactggcc ggccctcggg agtggcagac
     3661 ctgggaaatc gacttcctga aacagcaagc cgtggtgatg ggcatcgcca tccagcagag
     3721 ctagaagctt ggatccaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     3781 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3841 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     3901 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3961 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4021 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     4081 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     4141 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     4201 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4261 gcctcttccg cgtcttcgag atctgcctcg actgtgcctt ctagttgcca gccatctgtt
     4321 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     4381 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     4441 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggac
     4501 tcgagttaag ggcgaattcc cgataaggat cttcctagag catggctacg tagataagta
     4561 gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc
     4621 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     4681 tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta attcactggc
     4741 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4801 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4861 ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     4921 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     4981 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     5041 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     5101 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     5161 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     5221 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     5281 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     5341 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5401 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5461 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5521 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5581 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5641 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5701 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5761 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5821 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5881 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5941 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     6001 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     6061 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     6121 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6181 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6241 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6301 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6361 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6421 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6481 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6541 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6601 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6661 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6721 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6781 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6841 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6901 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6961 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     7021 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     7081 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     7141 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     7201 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     7261 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     7321 attcattaat gcagctg
//
