LOCUS       sequence-359597-        5258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible mammalian expression vector for ZsGreen1-tagged HP1alpha.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5258)
  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 5258)
  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: pTRE3G-ZSGreen1-mHP1alpha
FEATURES             Location/Qualifiers
     source          1..5258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        4..382
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    12..30
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    48..66
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    84..102
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    120..138
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    156..174
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    192..210
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    228..246
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     291..315
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             389..1084
                     /codon_start=1
                     /product="Zoanthus green fluorescent protein (Matz et al., 
                     1999)"
                     /label=ZsGreen
                     /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN
                     LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIADYFKNSCPAGYTWDRSFLFEDGAV
                     CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG
                     DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH
                     AIASGSALP"
     misc_feature    join(1092..1667,1668..1670,1671..1679)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1260..1277)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1487..1506
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     intron          2410..2475
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             2605..2625
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    3050..3171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3087..3106)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3141..3160
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3257..3274)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3428..4016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3508..3527)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4187..4978,4979..5047))
                     /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     4810..4829
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5048..5152)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5220..5238
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagttta ctccctatca gtgatagaga acgtatgaag agtttactcc ctatcagtga
       61 tagagaacgt atgcagactt tactccctat cagtgataga gaacgtataa ggagtttact
      121 ccctatcagt gatagagaac gtatgaccag tttactccct atcagtgata gagaacgtat
      181 ctacagttta ctccctatca gtgatagaga acgtatatcc agtttactcc ctatcagtga
      241 tagagaacgt ataagcttta ggcgtgtacg gtgggcgcct ataaaagcag agctcgttta
      301 gtgaaccgtc agatcgcctg gagcaattcc acaacacttt tgtcttatac caactttccg
      361 taccacttcc taccctcgta aagtcgacat ggctcagtca aagcacggtc taacaaaaga
      421 aatgacaatg aaataccgta tggaagggtg cgtcgatgga cataaatttg tgatcacggg
      481 agagggcatt ggatatccgt tcaaagggaa acaggctatt aatctgtgtg tggtcgaagg
      541 tggaccattg ccatttgccg aagacatatt gtcagctgcc tttatgtacg gaaacagggt
      601 tttcactgaa tatcctcaag acatagctga ctatttcaag aactcgtgtc ctgctggtta
      661 tacatgggac aggtcttttc tctttgagga tggagcagtt tgcatatgta atgcagatat
      721 aacagtgagt gttgaagaaa actgcatgta tcatgagtcc aaattttatg gagtgaattt
      781 tcctgctgat ggacctgtga tgaaaaagat gacagataac tgggagccat cctgcgagaa
      841 gatcatacca gtacctaagc aggggatatt gaaaggggat gtctccatgt acctccttct
      901 gaaggatggt gggcgtttac ggtgccaatt cgacacagtt tacaaagcaa agtctgtgcc
      961 aagaaagatg ccggactggc acttcatcca gcataagctc acccgtgaag accgcagcga
     1021 tgctaagaat cagaaatggc atctgacaga acatgctatt gcatccggat ctgcattgcc
     1081 ctgacggccg gcccctctcc ctcccccccc ccctaacgtt actggccgaa gccgcttgga
     1141 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     1201 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     1261 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     1321 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     1381 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     1441 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     1501 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     1561 ggggcctcgg tacacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     1621 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaaccg
     1681 ggccggatat cacgcgtatg ggaaagaaga ccaagaggac agccgacagc tcttcttcag
     1741 aggatgagga ggaatatgtg gtggaaaagg tgttggacag gcgcatggtt aaggggcaag
     1801 tggaatatct gttgaagtgg aaaggctttt ctgaggagca caatacttgg gaacctgaga
     1861 agaacttgga ttgtcctgaa ctaatttctg agtttatgaa aaagtataag aagatgaagg
     1921 agggtgaaaa caataagccc agggagaaat cagaaggaaa caagaggaaa tccagtttct
     1981 ccaacagcgc tgatgatatt aaatctaaaa aaaagagaga gcaaagcaat gatatcgctc
     2041 ggggctttga gagaggactg gaaccagaaa agatcatcgg agcaacagat tcctgcggtg
     2101 acttaatgtt cttaatgaaa tggaaagaca cagatgaagc tgacctggtt cttgcaaaag
     2161 aagctaacgt gaagtgtcca cagattgtga tagcatttta tgaagagaga ctgacgtggc
     2221 acgcatatcc agaggatgcg gaaaacaaag aaaaagaaag cgcgaagagc taaggatcca
     2281 atgtaactgt attcagcgat gacgaaattc ttagctattg taatactcta gaggatcttt
     2341 gtgaaggaac cttacttctg tggtgtgaca taattggaca aactacctac agagatttaa
     2401 agctctaagg taaatataaa atttttaagt gtataatgtg ttaaactact gattctaatt
     2461 gtttgtgtat tttagattcc aacctatgga actgatgaat gggagcagtg gtggaatgcc
     2521 tttaatgagg aaaacctgtt ttgctcagaa gaaatgccat ctagtgatga tgaggctact
     2581 gctgactctc aacattctac tcctccaaaa aagaagagaa aggtagaaga ccccaaggac
     2641 tttccttcag aattgctaag ttttttgagt catgctgtgt ttagtaatag aactcttgct
     2701 tgctttgcta tttacaccac aaaggaaaaa gctgcactgc tatacaagaa aattatggaa
     2761 aaatattctg taacctttat aagtaggcat aacagttata atcataacat actgtttttt
     2821 cttactccac acaggcatag agtgtctgct attaataact atgctcaaaa attgtgtacc
     2881 tttagctttt taatttgtaa aggggttaat aaggaatatt tgatgtatag tgccttgact
     2941 agagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3001 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3061 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3121 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3181 cggctctaga gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3241 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3301 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     3361 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     3421 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3481 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3541 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3601 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3661 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3721 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3781 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3841 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3901 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3961 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4021 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4081 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4141 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4201 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4261 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4321 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4381 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4441 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4501 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4561 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4621 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4681 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4741 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4801 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4861 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4921 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4981 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5041 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5101 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5161 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     5221 ataggcgtat cacgaggccc tttcgtcttc aagaattc
//
