LOCUS       sequence-235185-        7032 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET biosensor. To monitor histone H3 lysine 9 tri-methylation in 
            mammalian cells by FRET.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7032)
  AUTHORS   Peng Q, Lu S, Shi Y, Pan Y, Limsakul P, Chernov AV, Qiu J, Chai X, 
            Shi Y, Wang P, Ji Y, Li YJ, Strongin AY, Verkhusha VV, Izpisua 
            Belmonte JC, Ren B, Wang Y, Chien S, Wang Y
  TITLE     Coordinated histone modifications and chromatin reorganization in a 
            single cell revealed by FRET biosensors.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Nov 26. pii: 1811818115. doi: 
            10.1073/pnas.1811818115.
  PUBMED    30478057
REFERENCE   2  (bases 1 to 7032)
  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. 2018 Nov 26. pii: 1811818115. doi: 
            10.1073/pnas.1811818115."
COMMENT     Sequence Label: H3K9me3 biosensor
FEATURES             Location/Qualifiers
     source          1..7032
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..665
                     /label=chicken beta-actin promoter
     intron          666..1686
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1606..1628
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1694..1713
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(2950..2952,2953..2955,2956..3468)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     primer_bind     complement(2995..3016)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3256..3275)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3603..3624
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      3628..3637
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4054..4073)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4119..4174
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4173..4192)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4535..4551)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4535..4551)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4548..4570)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4559..4575
                     /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)."
     promoter        complement(join(4583..4589,4590..4607,4608..4613))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4628..4649
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4708..4903
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4754..4889
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4909..5043
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4946..4965)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5000..5019
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5111..5128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5282..5870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5362..5381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6041..6832,6833..6901))
                     /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     6664..6683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6902..7006)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggg gcgcgccagg cggggcgggg cggggcgagg
      541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
      721 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
      781 ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc
      841 gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg
      901 cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg
      961 agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg
     1021 ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg
     1081 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc
     1141 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
     1201 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
     1261 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
     1321 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     1381 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
     1441 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
     1501 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
     1561 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1621 gtgtgaccgg cggctctagc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     1681 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattc
     1741 atgtctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga attagatggt
     1801 gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc tacgtacggt
     1861 aaattgacct taaaattact ctgtactact ggtaaattgc cagttccatg gccaacctta
     1921 gtcactactt taggttatgg tgttcaatgt tttgctagat acccagatca tatgaaacaa
     1981 catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac tatttttttc
     2041 aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga taccttagtt
     2101 aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt aggtcacaaa
     2161 ttggaataca actataactc tcacaatgtt tacatcactg ctgacaaaca aaagaatggt
     2221 atcaaagcta acttcaaaat tagacacaac attgaagatg gtggtgttca attagctgac
     2281 cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga caaccattac
     2341 ttatcctatc aatctgcctt attcaaagat ccaaacgaaa agagagatca catggtcttg
     2401 ttagaatttt tgactgctgc tgaggaggag tacgccgtgg aaaagatcat cgacaggcgg
     2461 gtgcgcaagg gaaaggtgga gtactatctg aaatggaagg gctatcccga aactgagaac
     2521 acgtgggagc cggagaacaa tctcgactgc caggatctta tccagcagta cgaggcgagc
     2581 cgcaaggatg ggtctagcgc aggcggatca gctggagggt ctgcaggggg tagtgcaggt
     2641 ggctcagctg gcgggagcgg ctcagctggg ggatctgctg gtggcagtac ctcagcaggc
     2701 ggtagcgccg gaggttctgc tggtggctcc gcaggagggt ctgcaggcgg ttccgggagt
     2761 gcaggtggat ctgcaggtgg gtcaacaagt gctggtggat ccgcaggagg ttcagcaggc
     2821 gggagtgctg gaggctctgc aggcggtagc gggagtgccg gtggcagcgc agggggaagc
     2881 actagtgctg gaggcagtgc aggtgacagc gcaggaggct ctgccggggg aagcgccggg
     2941 ggcgggacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3001 gagctggacg gcgacgtaaa cggccacagg ttcagcgtgt ccggcgaggg cgagggcgat
     3061 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     3121 tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt gcttcagccg ctaccccgac
     3181 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgt
     3241 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     3301 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3361 ctggggcaca agctggagta caactacatc agccacaacg tctatatcac cgccgacaag
     3421 cagaagaacg gcatcaaggc ccacttcaag atccgccaca acatcgagga cggcagcgtg
     3481 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3541 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3601 cacatggtcc tgctggagtt cgtgaccgcc gccatggcac gaaccaagca gactgcccgc
     3661 aaatccaccg gcggtaaggc accacggaag cagctggcta ccaaagctgc tagaaagagt
     3721 gctcctgcaa cagggggagt gaaaaagcct caccgatatc gccctgggac tgtggccctt
     3781 agggagatca gaagatacca aaaaagcaca gagttgctga tccggaagct gccatttcaa
     3841 cgactggtgc gggagatagc ccaggacttt aagaccgatc ttaggttcca atcctcagct
     3901 gttatggcac tccaggaggc cagtgaagct tacctcgttg ggttgtttga agacaccaac
     3961 ctgtgcgcca tccacgccaa aagggtgaca atcatgccga aggatattca gctcgccaga
     4021 agaatcagag gggagagggc ttaggtcgac ggatcttttt ccctctgcca aaaattatgg
     4081 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     4141 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     4201 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     4261 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     4321 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     4381 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     4441 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     4501 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4561 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4621 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4681 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     4741 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4801 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4861 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     4921 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4981 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     5041 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5101 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5161 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5221 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5281 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5341 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5401 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5461 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5521 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5581 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5641 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5701 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5761 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5821 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5881 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5941 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6001 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6061 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6121 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6181 gtgacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6241 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6301 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6361 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6421 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6481 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6541 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6601 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6661 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6721 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6781 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6841 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6901 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6961 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7021 aagtgccacc tg
//
