LOCUS       sequence-449672-        4982 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Human H2B K120C mutant in bacterial cells for fluorescent 
            labelling of histone octamers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4982)
  AUTHORS   Uckelmann M, Levina V, Taveneau C, Han Ng X, Pandey V, Martinez J, 
            Mendiratta S, Houx J, Boudes M, Venugopal H, Trepout S, Zhang Q, 
            Flanigan S, Li M, Sierecki E, Gambin Y, Das PP, Bell O, de Marco A, 
            Davidovich C
  TITLE     Dynamic PRC1-CBX8 stabilizes a porous structure of chromatin 
            condensates.
  JOURNAL   bioRxiv [Preprint]. 2024 Feb 12:2023.05.08.539931. doi: 
            10.1101/2023.05.08.539931.
  PUBMED    38405976
REFERENCE   2  (bases 1 to 4982)
  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"; journalName: "bioRxiv 
            [Preprint]. 2024 Feb 12:2023.05.08.539931. doi: 
            10.1101/2023.05.08.539931."
COMMENT     Sequence Label: pET3a.H2B-K120C
FEATURES             Location/Qualifiers
     source          1..4982
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    775..914
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(840..859)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1000..1022
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1016..1207)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     2665..2684
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2768..2787
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2768..2786
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2818..2840
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(3279..3297)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3293..3340
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(3706..3711,3712..3728,3729..3734))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(3761..3779)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3847..3951
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3952..4020,4021..4812)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4170..4189)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
      841 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      901 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
      961 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1021 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     1081 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     1141 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     1201 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     1261 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     1321 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     1381 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     1441 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     1501 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     1561 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     1621 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg
     1681 acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata
     1741 tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa
     1801 ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc
     1861 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac
     1921 aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag
     1981 cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg
     2041 atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc
     2101 ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa
     2161 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     2221 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     2281 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     2341 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     2401 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     2461 catcggtcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     2521 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     2581 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     2641 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     2701 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     2761 gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag aaataatttt
     2821 gtttaacttt aagaaggaga tatacatatg ccagaaccag cgaaatcggc acctgcaccg
     2881 aaaaaaggta gcaaaaaagc agttaccaaa gcccagaaaa aagacgggaa aaaacgtaaa
     2941 cgtagccgca aagaaagcta tagcgtgtat gtttacaaag tgctgaaaca ggttcatccg
     3001 gataccggca ttagcagcaa agcaatgggt attatgaata gcttcgtgaa cgatatcttt
     3061 gaacgtattg ccggtgaagc aagccgtctg gcacattata acaaacgtag caccattacc
     3121 agccgtgaaa ttcagaccgc agttcgtctg ctgctgcctg gtgaactggc aaaacatgca
     3181 gttagcgaag gcaccaaagc cgtgacctgc tataccagca gcaaataagg atccggctgc
     3241 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     3301 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     3361 cggatatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta
     3421 gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg agaacgggtg
     3481 cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga ctggcgatgc
     3541 tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc
     3601 ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac
     3661 acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa agcttatcga
     3721 tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat acgcctattt
     3781 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     3841 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     3901 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     3961 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4021 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4081 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4141 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     4201 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4261 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4321 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4381 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     4441 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     4501 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     4561 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     4621 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     4681 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     4741 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     4801 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     4861 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4921 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4981 tc
//
