LOCUS       sequence-359696-        6801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescently tagged PMLIII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6801)
  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 6801)
  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-hPMLIII
FEATURES             Location/Qualifiers
     source          1..6801
                     /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    3589..3710
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3626..3645)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3680..3699
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3717..4172)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3854..3875)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4066..4085
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4199..4303
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4305..4662
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4305..4496
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4328..4348)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4513..4648
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4697..5491
                     /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             4697..5491
                     /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(4751..4770)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5361..5380
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5679..5698)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5723..5770
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6099..6687
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6588..6607
                     /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 ctgtacaagt ccggactcag atctaaaccg agaatcgaaa ctaagctggg gtccatggag
     1381 cctgcacccg cccgatctcc gaggccccag caggaccccg cccggcccca ggagcccacc
     1441 atgcctcccc ccgagacccc ctctgaaggc cgccagccca gccccagccc cagccctaca
     1501 gagcgagccc ccgcttcgga ggaggagttc cagtttctgc gctgccagca atgccaggcg
     1561 gaagccaagt gcccgaagct gctgccttgt ctgcacacgc tgtgctcagg atgcctggag
     1621 gcgtcgggca tgcagtgccc catctgccag gcgccctggc ccctaggtgc agacacaccc
     1681 gccctggata acgtcttttt cgagagtctg cagcggcgcc tgtcggtgta ccggcagatt
     1741 gtggatgcgc aggctgtgtg cacccgctgc aaagagtcgg ccgacttctg gtgctttgag
     1801 tgcgagcagc tcctctgcgc caagtgcttc gaggcacacc agtggttcct caagcacgag
     1861 gcccggcccc tagcagagct gcgcaaccag tcggtgcgtg agttcctgga cggcacccgc
     1921 aagaccaaca acatcttctg ctccaacccc aaccaccgca cccctacgct gaccagcatc
     1981 tactgccgag gatgttccaa gccgctgtgc tgctcgtgcg cgctccttga cagcagccac
     2041 agtgagctca agtgcgacat cagcgcagag atccagcagc gacaggagga gctggacgcc
     2101 atgacgcagg cgctgcagga gcaggatagt gcctttggcg cggttcacgc gcagatgcac
     2161 gcggccgtcg gccagctggg ccgcgcgcgt gccgagaccg aggagctgat ccgcgagcgc
     2221 gtgcgccagg tggtagctca cgtgcgggct caggagcgcg agctgctgga ggctgtggac
     2281 gcgcggtacc agcgcgacta cgaggagatg gccagtcggc tgggccgcct ggatgctgtg
     2341 ctgcagcgca tccgcacggg cagcgcgctg gtgcagagga tgaagtgcta cgcctcggac
     2401 caggaggtgc tggacatgca cggtttcctg cgccaggcgc tctgccgcct gcgccaggag
     2461 gagccccaga gcctgcaagc tgccgtgcgc accgatggct tcgacgagtt caaggtgcgc
     2521 ctgcaggacc tcagctcttg catcacccag gggaaagatg cagctgtatc caagaaagcc
     2581 agcccagagg ctgccagcac tcccagggac cctattgacg ttgacctgcc cgaggaggca
     2641 gagagagtga aggcccaggt tcaggccctg gggctggctg aagcccagcc tatggctgtg
     2701 gtacagtcag tgcccggggc acaccccgtg ccagtgtacg ccttctccat caaaggccct
     2761 tcctatggag aggatgtctc caatacaacg acagcccaga agaggaagtg cagccagacc
     2821 cagtgcccca ggaaggtcat caagatggag tctgaggagg ggaaggaggc aaggttggct
     2881 cggagctccc cggagcagcc caggcccagc acctccaagg cagtctcacc accccacctg
     2941 gatggaccgc ctagccccag gagccccgtc ataggaagtg aggtcttcct gcccaacagc
     3001 aaccacgtgg ccagtggcgc cggggaggca gaggaacgcg ttgtggtgat cagcagctcg
     3061 gaagactcag atgccgaaaa ctcggtctct tccagccctc agtctgaggt tctgtattgg
     3121 aaagtgcatg gagcccatgg agaccgccga gccacagtcc tcgccagccc actcctcgcc
     3181 agcccactcc tcgccagccc actcctcgcc agtccagtct ctgctgagag cacaaggagc
     3241 ctccagcctg ccctgtggca cataccaccc cccagcttgg cctccccacc agcccgctga
     3301 gcaggctgcc acccccgatg ctgagcctca cagcgagcct cctgatcacc aggagcgccc
     3361 tgccgtccac cgtgggatcc gctacctgtt gtacagagca cagagagcca tccgccttcg
     3421 ccatgccctc cgcttgcacc ctcaattgca tcgggcccgg gatccaccgg atctagataa
     3481 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3541 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3601 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3661 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat
     3721 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3781 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3841 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3901 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3961 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4021 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4081 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4141 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     4201 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4261 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     4321 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4381 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4441 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4501 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4561 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4621 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     4681 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4741 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4801 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4861 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4921 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4981 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5041 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5101 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5161 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5221 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5281 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5341 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5401 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5461 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5521 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5581 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5641 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     5701 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     5761 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5821 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5881 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5941 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6001 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6061 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6121 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6181 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6241 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6301 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6361 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6421 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6481 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6541 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6601 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6661 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6721 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6781 aaccgtatta ccgccatgca t
//
