LOCUS       sequence-367772-        5800 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP tagged H3K9me3 recognition motif in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5800)
  AUTHORS   Mendonca A, Sanchez O, Zhao H, Lin L, Min A, Yuan C
  TITLE     Development and application of novel BiFC probes for cell sorting 
            based on epigenetic modification.
  JOURNAL   Cytometry A. 2022 Apr;101(4):339-350. doi: 10.1002/cyto.a.24530. 
            Epub 2022 Jan 9.
  PUBMED    35001539
REFERENCE   2  (bases 1 to 5800)
  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"; doi: 
            "10.1002/cyto.a.24530"; journalName: "Cytometry A"; date: "2022-04";
            volume: "101"; issue: "4"; pages: "339-350"
COMMENT     Sequence Label: H3K9me3Probe_FullLength
FEATURES             Location/Qualifiers
     source          1..5800
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             696..716
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1311..1320
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1317..1319,1320..1322,1323..2036)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1362..1383)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1623..1642)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1970..1991
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      2065..2074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2078..2212
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2115..2134)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2169..2188
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2281..2638
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2281..2472
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2304..2324)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2489..2624
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2658..2675)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2658..2674)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2667..2689)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2887..3342
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2974..2993)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3184..3205
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3359..3378)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3513..3535
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3573..3591)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3659..3763
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3764..3832,3833..4624)
                     /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(3982..4001)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4795..5383
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5284..5303
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5537..5554
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5671..5692
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5707..5712,5713..5730,5731..5737)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5745..5761
                     /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)."
     primer_bind     5750..5772
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5769..5785
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5769..5785
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggcccg ggatgccaaa aaagaagaga aaggtaggta
      721 gcggcagcgg tagcggttcc ggaggcagcg aattcgcttc ccaggagttt gaggttgaag
      781 ctattgttga caaaagacag gataaaaatg ggaatacaca gtatttggtt cggtggaaag
      841 gttatgacaa acaggatgac acttgggaac cagagcagca cctcatgaac tgtgaaaaat
      901 gtgtacatga ttttaataga cgacagactg aaaaacagaa aaaactgaca tggactacaa
      961 ccagtagaat tttttcaaac aatgccggtt ccggaggtgg aggtggttcc caattcgctt
     1021 cccaggagtt tgaggttgaa gctattgttg acaaaagaca ggataaaaat gggaatacac
     1081 agtatttggt tcggtggaaa ggttatgaca aacaggatga cacttgggaa ccagagcagc
     1141 acctcatgaa ctgtgaaaaa tgtgtacatg attttaatag acgacagact gaaaaacaga
     1201 aaaaactgac atggactaca accagtagaa ttttttcaaa caatgccggt tccggaggtg
     1261 gaggtggttc ccaattgggt tccggaggtg tcgacagctg cctaccggtc gccaccatgg
     1321 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1381 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1441 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1501 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     1561 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1621 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1681 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     1741 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     1801 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     1861 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     1921 tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     1981 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaatcct
     2041 ctagagtcga cctgcagaag cttggccgcc atggcccaac ttgtttattg cagcttataa
     2101 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2161 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tcgggaatta
     2221 attcggcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt
     2281 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     2341 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2401 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2461 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     2521 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     2581 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     2641 gctgttaaca gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     2701 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     2761 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     2821 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     2881 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2941 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3001 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3061 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     3121 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3181 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg
     3241 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3301 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     3361 caatctgctc tgatgccgca tagttaagcc wssymcraym csckcyrayr ccssmtrryk
     3421 mrgccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3481 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3541 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3601 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3661 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3721 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3781 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3841 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     3901 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3961 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4021 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4081 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4141 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4201 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4261 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4321 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4381 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4441 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4501 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4561 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4621 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4681 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4741 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4801 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4861 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4921 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4981 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5041 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5101 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5161 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5221 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5281 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5341 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5401 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5461 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5521 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     5581 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     5641 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     5701 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     5761 atttcacaca ggaaacagct atgacatgat tacgaattaa
//
