LOCUS       sequence-359688-        6318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescently tagged HP1alpha fused to the arginine/glycine-rich RGG
            domain of LAF-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6318)
  AUTHORS   Erdel F, Rademacher A, Vlijm R, Tunnermann J, Frank L, Weinmann R, 
            Schweigert E, Yserentant K, Hummert J, Bauer C, Schumacher S, Al 
            Alwash A, Normand C, Herten DP, Engelhardt J, Rippe K
  TITLE     Mouse Heterochromatin Adopts Digital Compaction States without 
            Showing Hallmarks of HP1-Driven Liquid-Liquid Phase Separation.
  JOURNAL   Mol Cell. 2020 Apr 16;78(2):236-249.e7. doi: 
            10.1016/j.molcel.2020.02.005. Epub 2020 Feb 25.
  PUBMED    32101700
REFERENCE   2  (bases 1 to 6318)
  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: 
            "10.1016/j.molcel.2020.02.005"; journalName: "Mol Cell"; date: 
            "2020-04-16- 16"; volume: "78"; issue: "2"; pages: "236-249.e7"
COMMENT     Sequence Label: pRGG-GFP-RGG-mHP1alpha
FEATURES             Location/Qualifiers
     source          1..6318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1135..1155
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(1156..1158,1159..1161,1162..1872)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1201..1222)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     polyA_signal    3106..3227
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3143..3162)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3197..3216
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3234..3689)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3371..3392)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3583..3602
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3716..3820
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3822..4179
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3822..4013
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3845..3865)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4030..4165
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4214..5008
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4214..5008
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4268..4287)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4878..4897
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5196..5215)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5240..5287
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5616..6204
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6105..6124
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggaatc gaatcaatct aacaacggtg gatcgggaaa cgcagcattg
      661 aatcgtgggg gccgttacgt accaccacac ttgagaggcg gagatggtgg tgccgccgct
      721 gctgcctcag ccggaggcga cgacagacgc ggtggggccg ggggtggtgg ctatcgtaga
      781 ggtggtggaa attcaggggg cggcggcgga ggaggctacg atcgtggata caacgacaac
      841 cgggatgacc gtgacaatcg gggcggctcc ggcggttatg gccgggacag aaattacgag
      901 gatcgcggtt ataacggggg tggtggggga ggaggcaatc gcggctacaa taataaccgc
      961 ggtgggggtg gaggcggcta taatcggcag gaccgtggag acggcggttc tagtaatttc
     1021 tcgcggggag ggtataacaa cagagatgag ggctcagaca acagaggctc aggcagatca
     1081 tacaacaacg atcggcgcga caatggtggt gacgggggcg gaggcggttc cctcgagaat
     1141 ttatatttcc agggcatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1201 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1261 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1321 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgatat
     1381 ccagatcata tgaagcagca cgatttcttt aaaagtgcga tgcccgaggg ttatgttcag
     1441 gagcgcacca tcttcttcaa ggacgacggg aattataaga cccgcgctga ggttaagttt
     1501 gaaggggata cccttgttaa cagaattgag ctgaagggta tagactttaa agaagatggc
     1561 aatatattgg ggcacaagct cgaatataat tacaacagtc ataacgtata tatcatggct
     1621 gacaagcaaa agaatggcat aaaagtgaat tttaagatac gacacaatat agaggatggt
     1681 agcgtccagc ttgcggatca ttaccagcag aatacaccaa ttggtgacgg cccagtactg
     1741 ctgccggata atcactatct cagtacccag agtgcattgt ccaaagaccc gaacgagaaa
     1801 cgcgatcaca tggtactgct ggaatttgta acggctgctg gaattacatt gggaatggac
     1861 gagttgtaca aggaattcgg aggcggtgga agcgagtcta atcagagcaa taacgggggt
     1921 agtggcaatg ccgcgctcaa tcggggaggc cgctacgtgc ctcctcacct cagagggggc
     1981 gatgggggcg ccgctgccgc tgcgagcgcc ggtggcgatg atcggcgggg aggagcgggt
     2041 ggaggtggtt atcgccgagg tggcgggaac agtgggggtg gaggaggcgg tggatatgat
     2101 cgaggttaca acgacaatcg ggacgatcga gataacagag ggggttcagg tggatacgga
     2161 agagacagga attacgagga cagaggatac aatggcggag gcggtggggg aggtaataga
     2221 ggatacaaca ataatcgagg aggcgggggg ggagggtaca atcgccaaga ccgcggtgat
     2281 ggaggaagtt ctaactttag cagaggtgga tacaacaata gggatgaagg ctcagacaac
     2341 agaggttctg gtcgaagcta caataacgac agacgcgaca acggtggcga tgggtccgga
     2401 atgggaaaga agaccaagag gacagccgac agctcttctt cagaggatga ggaggaatat
     2461 gtggtggaaa aggtgttgga caggcgcatg gttaaggggc aagtggaata tctgttgaag
     2521 tggaaaggct tttctgagga gcacaatact tgggaacctg agaagaactt ggattgtcct
     2581 gaactaattt ctgagtttat gaaaaagtat aagaagatga aggagggtga aaacaataag
     2641 cccagggaga aatcagaagg aaacaagagg aaatccagtt tctccaacag cgctgatgat
     2701 attaaatcta aaaaaaagag agagcaaagc aatgatatcg ctcggggctt tgagagagga
     2761 ctggaaccag aaaagatcat cggagcaaca gattcctgcg gtgacttaat gttcttaatg
     2821 aaatggaaag acacagatga agctgacctg gttcttgcaa aagaagctaa cgtgaagtgt
     2881 ccacagattg tgatagcatt ttatgaagag agactgacgt ggcacgcata tccagaggat
     2941 gcggaaaaca aagaaaaaga aagcgcgaag agctaaggat ccaccggatc tagataactg
     3001 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     3061 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     3121 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3181 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     3241 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3301 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3361 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3421 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3481 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3541 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     3601 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     3661 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     3721 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3781 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     3841 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3901 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3961 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     4021 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     4081 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     4141 tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg
     4201 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4261 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4321 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4381 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4441 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4501 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4561 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4621 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4681 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4741 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4801 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4861 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4921 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4981 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa
     5041 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5101 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5161 ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg
     5221 gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg
     5281 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5341 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5401 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt
     5461 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5521 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5581 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5641 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5701 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5761 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtatc accgcctaca
     5821 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5881 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5941 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6001 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6061 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6121 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6181 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6241 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6301 cgtattaccg ccatgcat
//
