LOCUS       sequence-235136-        7034 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FRET biosensor. Disrupting HP1 binding capability in H3K9me3 
            biosensor to abolish the detection capability.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7034)
  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 7034)
  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: W45A mutant of H3K9me3 biosensor
FEATURES             Location/Qualifiers
     source          1..7034
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          220..1240
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1160..1182
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1248..1267
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             join(2504..2506,2507..2509,2510..3022)
                     /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(2549..2570)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2810..2829)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3157..3178
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      3182..3191
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3608..3627)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3673..3728
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3727..3746)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4089..4105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4089..4105)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4102..4124)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4113..4129
                     /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(4137..4143,4144..4161,4162..4167))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4182..4203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4262..4457
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4308..4443
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4463..4597
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4500..4519)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4554..4573
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4665..4682)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4836..5424)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4916..4935)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5595..6386,6387..6455))
                     /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     6218..6237
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6456..6560)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6595..6974
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(6976..7034,1..219)
                     /label=chicken beta-actin promoter
ORIGIN
        1 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
       61 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt
      121 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg
      181 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt
      241 cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg
      301 ttactcccac aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg
      361 gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag
      421 ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc
      481 gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt
      541 tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg
      601 gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg
      661 ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag
      721 cacggcccgg cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg
      781 ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag
      841 ggctcggggg aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc
      901 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
      961 atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg
     1021 aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc
     1081 cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg
     1141 gggggacggg gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagctagagc
     1201 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1261 tattgtgctg tctcatcatt ttggcaaaga attcatgtct aaaggtgaag aattattcac
     1321 tggtgttgtc ccaattttgg ttgaattaga tggtgatgtt aatggtcaca aattttctgt
     1381 ctccggtgaa ggtgaaggtg atgctacgta cggtaaattg accttaaaat tactctgtac
     1441 tactggtaaa ttgccagttc catggccaac cttagtcact actttaggtt atggtgttca
     1501 atgttttgct agatacccag atcatatgaa acaacatgac tttttcaagt ctgccatgcc
     1561 agaaggttat gttcaagaaa gaactatttt tttcaaagat gacggtaact acaagaccag
     1621 agctgaagtc aagtttgaag gtgatacctt agttaataga atcgaattaa aaggtattga
     1681 ttttaaagaa gatggtaaca ttttaggtca caaattggaa tacaactata actctcacaa
     1741 tgtttacatc actgctgaca aacaaaagaa tggtatcaaa gctaacttca aaattagaca
     1801 caacattgaa gatggtggtg ttcaattagc tgaccattat caacaaaata ctccaattgg
     1861 tgatggtcca gtcttgttac cagacaacca ttacttatcc tatcaatctg ccttattcaa
     1921 agatccaaac gaaaagagag atcacatggt cttgttagaa tttttgactg ctgctgagga
     1981 ggagtacgcc gtggaaaaga tcatcgacag gcgggtgcgc aagggaaagg tggagtacta
     2041 tctgaaagcc aagggctatc ccgaaactga gaacacgtgg gagccggaga acaatctcga
     2101 ctgccaggat cttatccagc agtacgaggc gagccgcaag gatgggtcta gcgcaggcgg
     2161 atcagctgga gggtctgcag ggggtagtgc aggtggctca gctggcggga gcggctcagc
     2221 tgggggatct gctggtggca gtacctcagc aggcggtagc gccggaggtt ctgctggtgg
     2281 ctccgcagga gggtctgcag gcggttccgg gagtgcaggt ggatctgcag gtgggtcaac
     2341 aagtgctggt ggatccgcag gaggttcagc aggcgggagt gctggaggct ctgcaggcgg
     2401 tagcgggagt gccggtggca gcgcaggggg aagcactagt gctggaggca gtgcaggtga
     2461 cagcgcagga ggctctgccg ggggaagcgc cgggggcggg accatggtga gcaagggcga
     2521 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2581 caggttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     2641 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2701 ctggggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2761 gtccgccatg cccgaaggct acgtccagga gcgtaccatc ttcttcaagg acgacggcaa
     2821 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2881 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     2941 catcagccac aacgtctata tcaccgccga caagcagaag aacggcatca aggcccactt
     3001 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     3061 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     3121 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     3181 cgccgccatg gcacgaacca agcagactgc ccgcaaatcc accggcggta aggcaccacg
     3241 gaagcagctg gctaccaaag ctgctagaaa gagtgctcct gcaacagggg gagtgaaaaa
     3301 gcctcaccga tatcgccctg ggactgtggc ccttagggag atcagaagat accaaaaaag
     3361 cacagagttg ctgatccgga agctgccatt tcaacgactg gtgcgggaga tagcccagga
     3421 ctttaagacc gatcttaggt tccaatcctc agctgttatg gcactccagg aggccagtga
     3481 agcttacctc gttgggttgt ttgaagacac caacctgtgc gccatccacg ccaaaagggt
     3541 gacaatcatg ccgaaggata ttcagctcgc cagaagaatc agaggggaga gggcttaggt
     3601 cgacggatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
     3661 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
     3721 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
     3781 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     3841 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     3901 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     3961 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     4021 cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg
     4081 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4141 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4201 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     4261 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4321 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4381 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4441 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     4501 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     4561 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     4621 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4681 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4741 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4801 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4861 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4921 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4981 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5041 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5101 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5161 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5221 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5281 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5341 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5401 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5461 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5521 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5581 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5641 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5701 ttaccatctg gccccagtgc tgcaatgata ccgcgtgacc cacgctcacc ggctccagat
     5761 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5821 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5881 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5941 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     6001 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     6061 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     6121 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     6181 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     6241 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     6301 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6361 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6421 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6481 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6541 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc gatcgacatt
     6601 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
     6661 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
     6721 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
     6781 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
     6841 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
     6901 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     6961 tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc
     7021 cctccccacc ccca
//
