LOCUS       sequence-359692-        7443 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescently tagged HP1alpha fused to PGL-1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7443)
  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 7443)
  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: pGFP-PGL-1-mHP1alpha
FEATURES             Location/Qualifiers
     source          1..7443
                     /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             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4231..4352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4268..4287)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4322..4341
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4359..4814)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4496..4517)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4708..4727
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4841..4945
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4947..5304
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4947..5138
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4970..4990)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5155..5290
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5339..6133
                     /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             5339..6133
                     /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(5393..5412)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6003..6022
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6321..6340)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6365..6412
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6741..7329
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7230..7249
                     /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 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaaga tggaggctaa caagcgagaa attgtggatt tcggtgggct gagatcctac
     1381 tttttcccaa atttggccca ctacatcacg aagaatgacg aagaactgtt caacaataca
     1441 tctcaggcca ataagttggc tgctttcgtg ttgggagctt ccaaagacgc tcctggagac
     1501 gaagacattt tggagatgat cttgccgaac gacgctaacg ctgccgttat tgctgctggc
     1561 atggacgttt gtcttcttct cggagataaa tttcgtccga aatttgatgc tgctgctgag
     1621 aaattgtcag gattgggaca tgctcatgat ctggtttctg tcatcgatga cgacaagaag
     1681 ctcggaatgc tcgcccgtaa ggccaagctg aagaagactg aagacgctaa gattcttcaa
     1741 gctcttctca aagtaatcgc aatcgatgat gccgccgaaa agtttgttga gctcacggaa
     1801 cttgtcagcc aactggatct ggactttgat gtttatgtcc taactaaaat cctcggactt
     1861 atctctgagg agactagcga cgaggtcgac attatccgtg ataacgtcgt caacgcattt
     1921 gatagctgca agccgttgtt gaaacagttg atgttggatg gtccgaaatc tgaacctgcc
     1981 gatccattca ttagtctcct catggatcca ctcgaagagt ctgttgggaa agttgtgaac
     2041 catatcgctc aacttttcga ggaagcgagt aaaaatgaag gagacgagtc acttgtcttg
     2101 agatctcaac tcggatatca gctctttttc cttatcgttc gttcacttgc agatggaaag
     2161 cgcgaggttt ccaagaaaat cctctctggg attcctacga gtgtcagagc ggaagtcttt
     2221 ccaggattgc aaagaagtgt ctacaagagt gcggtgttcc tcggcaatca catcatccaa
     2281 gttcttttag gatctaagaa gtcttttgag gattgggatg ttgttggagt cgcgaaagac
     2341 ttggagagcg cctggaaaag acgagccatt gcggaactta tcaagaagtt ccaagtctca
     2401 attctagagc aatgcttcga taagcccgtt ccactgattc ctcagagtcc actaaacaat
     2461 gatgcggtga tcgataatgt gaataaagct ctacagtttg cactgtggct tacagagttt
     2521 tacggatccg agaatgagac tgaagcgctg ggcgagcttc gtttcttgga ttccacttca
     2581 aaaaatcttc tcgtcgatag cttcaagaaa tttgttcaag gaatcaactc gaagactcat
     2641 gtcactcgaa ttgttgaatc actggagaag tgttgtttgt ccgatacacc aagtggcaga
     2701 aagagcaatg tgcaaccatc cacttcccaa cagcaggatt ccgcctacac gaaagaggaa
     2761 atgacgacgg ttcacaacac atacagcgta aatactaaag ctcaagttct caacggtctc
     2821 agtgatacca attcttccgg gttacttgtc gattcaaagg attcactcag tcttcaggag
     2881 atttcttgtg acgaagtgga ttcttcgact ttgctaagct catcaagaaa cattggagaa
     2941 ggtgttactg taaaagcggt cgatcctgtc ccagaaaagg ttaacgatgc acaacaacag
     3001 caaacagtta acgagatcga gatggcttcg gatgccaatc aagatacttc ttcaagtgct
     3061 tcgccagaag tagcgccaag tttttccacc gatggctggg attctccaac gaaatcagta
     3121 gctctgccac cgggcatgca gcagatagat gaagaggaaa ccacagttgc tgacaaagat
     3181 tctacaccac aaccacaagc tcgtgctgaa actgcatggg ggtcaggcga cgctactcca
     3241 atgcctcttc ctgccccgac taatcaatac aaagtcagtg gatttgggga agccaaagtt
     3301 gcaaaaggat tcggtcaatt tgctcccacc agttcagctt atggaggagg aggtggtcgc
     3361 ggaggatatg gcggtggaga ccgtggaggc agaggtggat acggcggaga tcgtggaggt
     3421 agaggtggat acggcggggg agaccgcgga ggacgaggtg gatacggcgg agatcgtgga
     3481 cgtggtggtt acgggggtcg tggtggacgc ggaggtttct ccggaatggg aaagaagacc
     3541 aagaggacag ccgacagctc ttcttcagag gatgaggagg aatatgtggt ggaaaaggtg
     3601 ttggacaggc gcatggttaa ggggcaagtg gaatatctgt tgaagtggaa aggcttttct
     3661 gaggagcaca atacttggga acctgagaag aacttggatt gtcctgaact aatttctgag
     3721 tttatgaaaa agtataagaa gatgaaggag ggtgaaaaca ataagcccag ggagaaatca
     3781 gaaggaaaca agaggaaatc cagtttctcc aacagcgctg atgatattaa atctaaaaaa
     3841 aagagagagc aaagcaatga tatcgctcgg ggctttgaga gaggactgga accagaaaag
     3901 atcatcggag caacagattc ctgcggtgac ttaatgttct taatgaaatg gaaagacaca
     3961 gatgaagctg acctggttct tgcaaaagaa gctaacgtga agtgtccaca gattgtgata
     4021 gcattttatg aagagagact gacgtggcac gcatatccag aggatgcgga aaacaaagaa
     4081 aaagaaagcg cgaagagcta aggatccacc ggatctagat aactgatcat aatcagccat
     4141 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4201 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4261 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4321 tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg ttaatatttt
     4381 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4441 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4501 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4561 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4621 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4681 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     4741 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     4801 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     4861 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4921 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     4981 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5041 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5101 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5161 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5221 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5281 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     5341 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5401 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5461 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5521 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5581 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5641 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     5701 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     5761 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     5821 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     5881 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     5941 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     6001 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     6061 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     6121 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     6181 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6241 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6301 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     6361 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     6421 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     6481 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     6541 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     6601 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6661 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6721 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6781 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6841 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6901 ccgtagttag gccaccactt caagaactct gtatcaccgc ctacatacct cgctctgcta
     6961 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7021 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7081 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7141 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7201 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7261 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7321 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7381 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     7441 cat
//
