LOCUS       sequence-486996-        5473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of green glucose sensor with inert mRuby3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5473)
  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 5473)
  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: pNoRBP.iGlucoSnFR2.mRuby3
FEATURES             Location/Qualifiers
     source          1..5473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     1296..1317
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1420..1441)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             2791..2808
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2874..2892)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2888..2935
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3032..3487
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3119..3138)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3329..3350
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3513..3617
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3618..3686,3687..4478)
                     /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(3836..3855)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4649..5237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5138..5157
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5391..5408
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgagatctaa gctggagatt
      121 ttctcttggt gggctgggga tgaaggccct gccctggagg cactcatcag actgtataaa
      181 cagaagtatc ctggagtaga ggtgataaac gccactgtca ccggaggtgc cggtgtaaat
      241 gctcgcgctg tccttaagac aaggatgctt ggtggtgatc ctcctgatac atttcaagtt
      301 gcagcaggga tggagttgat aggcacatgg gtggtggcta acagaatgga ggatctcagc
      361 gccttgtttc gacaagaggg ttggcttcaa gccttcccca agggccttat tgatctgatt
      421 agttataaag gtggtatatg gagtgtgccc gttaatattc acaggtcaaa cgtaatgtgg
      481 tatttgcctg caaagctaaa agagtgggga gttaaccccc cacggacttg ggatgagttt
      541 cttgccacct gtcaaacact taaacaaaag ggtctggaag ctccactggc tctcggggaa
      601 aattggacac aacaacacct ctgggagagt gtcgccttgg ctgtattggg gcctgatgat
      661 tggaataatc tgtggaatgg gaagctgaag ttcacagacc ctaaagcagt tagggcatgg
      721 gaagtctttg gccgcgtttt ggactgtgcc aacaaggatg cagcaggctt gtcttggcag
      781 caagctgtcg atcgggtagt acaggggaaa gccgctttta acgtaatggg cgactgggca
      841 gcaggatata tgactaccac tctcaagttg aagcctggta cagatttcgc atgggcacca
      901 agtcccggga cacagggcgt ctttatgatg gtttccgact ctttcggact tcccaaagga
      961 gccaaaaatc gacaaaacgc tatcaactgg cttcgattgg tcggctctaa agaaggtcaa
     1021 gatacattca acccactgaa agggtccatc gctgctcggc tcgatagcga cccaagcaag
     1081 tatccgtgta acgtctatat caccgccgac aagcagaaga acggcatcaa ggcgaacttc
     1141 aagatccgcc acaacgtgga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1201 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctggt gacccagtcc
     1261 gtgctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1321 gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg agggagcatg
     1381 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
     1441 gtaaacggcc acaagttcag cgtgcgcggc gagggcgagg gcgatgccac caacggcaag
     1501 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1561 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
     1621 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cagcttcaag
     1681 gacgacggca cctacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1741 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1801 gagtacaact ttaacaaccc caacgcatac ggacagagtg caatgcgaga ttggcggagt
     1861 aatcgcatcg ttggctccct ggtagcagga gcagtggccc ctgaaagctc catgtcacaa
     1921 ttcggaacag taatggaaat tttcctgcaa acacgcaacc cacaagcagc agcaaatgca
     1981 gcacaagcta ttgctgacca agttggcctg ggtcgccttg gtcagctgca gtctaccggc
     2041 tccggaaatg ctgtgggcca ggacacgcag gagcgtatgg tgtctaaggg cgaagagctg
     2101 atcaaggaaa atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg ccaccaattc
     2161 aaatgcacag gtgaaggaga aggcaggccg tacgagggag tgcaaaccat gaggatcaaa
     2221 gtcatcgagg gaggacccct gccatttgcc tttgacattc ttgccacgtc gttcatgtat
     2281 ggcagccgta cttttatcaa gtacccggca gacattcctg atttctttaa acagtccttt
     2341 cctgagggtt ttacttggga aagagttacg agatacgaag atggtggagt cgtcaccgtc
     2401 acgcaggaca ccagccttga ggatggcgaa ctcgtttaca acgtcaaagt cagaggggta
     2461 aactttccct ccaatggtcc cgtgatgcag aagaagacca agggttggga gcctaataca
     2521 gagatgatgt atccagcaga tggtggtctg aggggataca ctgatattgc actgaaagtt
     2581 gatggtggtg gccatctgca ttgcaatttc gtaacaactt acaggtcaaa aaagaccgtc
     2641 gggaacatca agatgcccgg tgtccatgcc gttgatcacc gcctggaaag gatagaggaa
     2701 agtgacaatg aaacgtacgt agtacaacgc gaagtcgcag ttgccaagta ctccaatctt
     2761 ggtggtggta tggatgaact ttataagtcc catcaccatc accaccatta agaattcgaa
     2821 gcttgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
     2881 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     2941 aggaggaact atatccggat ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3001 caacagttgc gcagcctgaa tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg
     3061 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     3121 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     3181 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     3241 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     3301 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     3361 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     3421 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     3481 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3541 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3601 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3661 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3721 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3781 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     3841 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3901 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3961 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4021 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4081 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     4141 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     4201 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     4261 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     4321 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     4381 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     4441 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     4501 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4561 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4621 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4681 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4741 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     4801 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     4861 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     4921 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     4981 gcacacagcc cagcttggag cgaacgacct acaccaaact gagataccta cagcgtgagc
     5041 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5101 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5161 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5221 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5281 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5341 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     5401 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     5461 ttcattaatg cag
//
