LOCUS       sequence-378448-        4718 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A binding-dead fluorescent biosensor for fructose 1,6-bisphosphate 
            (FBP) for negative control experiments (mammalian expression 
            vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4718)
  AUTHORS   Koberstein JN, Stewart ML, Smith CB, Tarasov AI, Ashcroft FM, Stork 
            PJS, Goodman RH
  TITLE     Monitoring glycolytic dynamics in single cells using a fluorescent 
            biosensor for fructose 1,6-bisphosphate.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Aug 2;119(31):e2204407119. doi: 
            10.1073/pnas.2204407119. Epub 2022 Jul 26.
  PUBMED    35881794
REFERENCE   2  (bases 1 to 4718)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2022-08-2"; volume: "119(31):e2204407119. 
            doi"; pages: " 10.1073/pnas.2204407119. Epub 2022 Jul 26"
COMMENT     Sequence Label: CMV-Hylight-T152E
FEATURES             Location/Qualifiers
     source          1..4718
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     94..113
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(332..436)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(511..530)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        702..1081
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1082..1285
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1236..1256
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1330..1349
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1330..1348
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1377..1386
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1695..1946
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGPVLLPDNH
                     YLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     1883..1904
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2010..2031)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(2961..2979)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2962..2979)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2999..3016)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3005..3229
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3430..4018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3510..3529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4189..4718,1..262,263..331))
                     /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"
ORIGIN
        1 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
       61 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
      121 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
      181 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
      241 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
      301 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
      361 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
      421 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
      481 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
      541 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
      601 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
      661 ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag
      721 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
      781 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      841 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      901 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      961 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
     1021 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1081 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1141 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1201 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1261 gtgggaggtc tatataagca gagctctctg gctaactaga gaacccactg cttactggct
     1321 tatcgaaatt aatacgactc actataggga gacccaagct tggtaccgag ctcggagcca
     1381 ccatgggcag caaggatgtt ttgggtttga ccttgttgga aaagactttg aaagaaagat
     1441 tgaacttgaa ggacgccatc atcgtttctg gtgattctga tcaatctcca tgggtcaaaa
     1501 aagaaatggg tagagctgct gttgcttgca tgaagaaaag attttctggt aagaacatcg
     1561 ttgctgttac tggtggtact gagattgaag ctgttgctga aatgatgacc ccagattcta
     1621 agaacagaga attgttgttt gttccagcta gaggtggttt gggtgaaccg ccttataacg
     1681 tctttatcat ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca
     1741 acatcgagga cggcggcgtg cagctcgcct atcactacca gcagaacacc cccatcggcg
     1801 acggccccgt gctgctgccc gacaaccact acctgagcgt gcagtccaaa ctgagcaaag
     1861 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1921 ctctcggcat ggacgagctg tacaagggcg gtaccggagg gagcatggtg agcaagggcg
     1981 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2041 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2101 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2161 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2221 agtccgccat gcccgaaggc tacattcagg agcgcaccat cttcttcaag gacgacggca
     2281 actataagac acgcgctgag gttaagttcg agggcgacac tctggttaac cgcatcgagc
     2341 tgaagggcat cgacttcaag gaggacggca acatcctggg ccataagctt gaatataact
     2401 tcaacaagga ggatgttaag aatcaagcta acaccatttg cgctcatatg gctgaaaaag
     2461 cttcaggtac ttacagattg ttattcgtcc caggtcaatt gtctcaaggt gcttactctt
     2521 ccattatcga agaaccatct gtcaaagaag tcttgaacac catcaaatcc gcttccatgt
     2581 tggttcacgg tattggtgaa gctaaaacta tggctcaacg cagaaacacc ccattggaag
     2641 atttgaaaaa gatcgatgat aacgatgctg ttaccgaagc tttcggttac tactttaatg
     2701 ctgatggtga agttgttcac aaggttcatt cagttggtat gcaattggat gatattgatg
     2761 ccatcccaga tattattgca gttgctggtg gttcttctaa ggctgaagca attgaagctt
     2821 acttcaagaa gccaagaaac actgttttgg ttactgatga aggtgctgcc aaaaagttgt
     2881 tgagagatga atcaggctaa gaattctgca gatatccatc acactggcgg ccgctcgagc
     2941 atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg ctgatcagcc
     3001 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3061 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3121 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3181 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3241 gaaagaacca gctggggctc tagtccggaa ctcgctgcgc tcggtcgttc ggctgcggcg
     3301 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3361 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3421 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3481 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3541 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3601 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3661 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3721 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3781 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3841 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3901 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3961 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4021 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4081 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4141 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4201 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4261 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4321 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4381 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4441 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4501 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4561 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4621 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4681 agcactgcat aattctctta ctgtcatgcc atccgtaa
//
