LOCUS       sequence-307934-        6023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human histone H2B with T116C mutation to facilitate 
            maleimide chemistry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6023)
  AUTHORS   Gibson BA, Doolittle LK, Schneider MWG, Jensen LE, Gamarra N, Henry 
            L, Gerlich DW, Redding S, Rosen MK
  TITLE     Organization of Chromatin by Intrinsic and Regulated Phase 
            Separation.
  JOURNAL   Cell. 2019 Oct 3;179(2):470-484.e21. doi: 
            10.1016/j.cell.2019.08.037. Epub 2019 Sep 19.
  PUBMED    31543265
REFERENCE   2  (bases 1 to 6023)
  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.cell.2019.08.037"; journalName: "Cell"; date: "2019-10-3-
            3"; volume: "179"; issue: "2"; pages: "470-484.e21"
COMMENT     Sequence Label: pET19b+H2B(T116C)
FEATURES             Location/Qualifiers
     source          1..6023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     80..99
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        80..98
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    99..123
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             138..160
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(604..622)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      618..665
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(840..845,846..862,863..868))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(895..913)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        981..1085
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1086..1154,1155..1946)
                     /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(1304..1323)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2117..2705
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2606..2625
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2859..2876
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2891..3030
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2956..2975)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3116..3138
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3132..3323)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5678..5697
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5717..5794)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6000..6019
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
       61 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
      121 tcccctctag aaataatttt gtttaacttt aagaaggaga tataccatgg gccctgagcc
      181 ggctaaatca gcgccagctc ctaagaaagg atctaaaaag gccgtcacaa aagcgcagaa
      241 gaaggacggc aaaaaacgta aacgttcccg caaagaatca tatagcattt atgtctataa
      301 agttctcaaa caagtgcacc cagatactgg catttctagt aaggcgatgg gtatcatgaa
      361 ttcatttgta aacgacattt ttgagcgcat tgccggcgaa gcctcccgtc tcgctcacta
      421 taataagcgt tcaacaatca cctcacgtga gattcaaact gcggtccggt tgctcttgcc
      481 aggcgaatta gctaaacatg cagtgtctga gggctgtaaa gcggtgacaa aatacacttc
      541 ggcgaaatga taaggatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
      601 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
      661 ttttgctgaa aggaggaact atatccggat atcccgcaag aggcccggca gtaccggcat
      721 aaccaagcct atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt
      781 gttagatttc atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca
      841 ttaaagctta tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg
      901 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
      961 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     1021 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     1081 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     1141 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     1201 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     1261 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     1321 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     1381 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     1441 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     1501 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     1561 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     1621 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     1681 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     1741 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     1801 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     1861 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     1921 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     1981 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     2041 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     2101 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     2161 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     2221 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     2281 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     2341 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     2401 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     2461 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     2521 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     2581 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     2641 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     2701 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     2761 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     2821 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     2881 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     2941 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg
     3001 ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg
     3061 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     3121 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa
     3181 agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc
     3241 tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg
     3301 gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg
     3361 gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat
     3421 gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag
     3481 agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag
     3541 ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc
     3601 cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc
     3661 gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc
     3721 taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg
     3781 cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct gctggagatg
     3841 gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag
     3901 aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca
     3961 ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt
     4021 atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa
     4081 tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc
     4141 cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg
     4201 gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg
     4261 tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct
     4321 gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca
     4381 agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct
     4441 cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga
     4501 cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt
     4561 tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat
     4621 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa ctgtcgtgcc agctgcatta
     4681 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     4741 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     4801 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     4861 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     4921 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     4981 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     5041 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     5101 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     5161 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     5221 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     5281 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     5341 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     5401 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     5461 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     5521 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     5581 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     5641 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     5701 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     5761 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     5821 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     5881 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     5941 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     6001 gtgatgtcgg cgatataggc gcc
//
