LOCUS       sequence-487004-        5647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of green glucose sensor (high affinity) with 
            inert HaloTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5647)
  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 5647)
  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.H348H.HaloTag
FEATURES             Location/Qualifiers
     source          1..5647
                     /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             2965..2982
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3048..3066)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3062..3109
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3206..3661
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3293..3312)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3503..3524
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3687..3791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3792..3860,3861..4652)
                     /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(4010..4029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4823..5411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5312..5331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5565..5582
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gatcacaacg 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 ggtacacgga gcagtggccc ctgaaagctc catgtcacaa
     1921 ttcggaacag taatggaaat tttcctgcaa acacgcaacc cacaagcagc agcaaatgca
     1981 gcacaagcta ttgctgacca agttggcctg ggtcgccttg gtcagctgca gggcggaggc
     2041 tcgggcggag gtgggtcggg tggcggcgga tcaatggccg agatcggcac cggcttcccc
     2101 ttcgaccccc actacgtgga ggtgctgggc gagcgcatgc actacgttga cgtgggcccc
     2161 cgcgacggca cccccgtgct gttcctgcac ggcaacccca ccagcagcta cgtgtggcgc
     2221 aacatcatcc cccacgtggc ccccacccac cgcactatcg cccccgacct gatcggcatg
     2281 ggcaagagcg acaagcccga cctgggctac ttcttcgacg accacgtgcg cttcatggac
     2341 gccttcatcg aggccctggg cctggaggag gtggtgctgg tgatccacga ctggggcagc
     2401 gccctgggct tccactgggc caagcgcaac cccgagcgcg tgaagggcat cgccttcatg
     2461 gagttcatcc gccccatccc cacctgggac gagtggcccg agttcgcccg cgagaccttc
     2521 caggccttcc gcaccaccga cgtgggccgc aagctgatca tcgaccagaa cgtgttcatc
     2581 gagggcaccc tgcccatggg cgtggtgcgc cccctgaccg aggtggagat ggaccactac
     2641 cgcgagccct tcctgaaccc cgtggaccgc gagcccctgt ggcgcttccc caacgagctg
     2701 cccatcgccg gcgagcccgc caacatcgtg gccctggtgg aggagtacat ggactggctg
     2761 caccagagcc ccgtgcccaa gctgctgttc tggggcaccc ccggcgtgct gatccccccc
     2821 gccgaggccg cccgcctggc caagagcctg cccaacgtta aggccgtgga catcggcccc
     2881 ggcctgaacc tgctccagga ggacaacccc gacctgatcg gcagcgagat cgcccgctgg
     2941 ctgagcaccc tcgagatcag cggccatcac catcaccacc attaagaatt cgaagcttga
     3001 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
     3061 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     3121 aactatatcc ggatctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3181 ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     3241 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     3301 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     3361 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     3421 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     3481 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     3541 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     3601 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     3661 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3721 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3781 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     3841 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     3901 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     3961 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     4021 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     4081 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     4141 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     4201 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     4261 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     4321 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     4381 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     4441 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     4501 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     4561 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     4621 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     4681 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4741 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5161 agcccagctt ggagcgaacg acctacacca aactgagata cctacagcgt gagctatgag
     5221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5461 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     5521 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     5581 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5641 aatgcag
//
