LOCUS       sequence-310150-        6023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human histone H2B.
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
FEATURES             Location/Qualifiers
     source          1..6023
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..46
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     130..149
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        130..148
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    149..173
                     /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             188..210
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(654..672)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      668..715
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(890..895,896..912,913..918))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(945..963)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1031..1135
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1136..1204,1205..1996)
                     /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(1354..1373)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2167..2755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2656..2675
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2909..2926
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2941..3080
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3006..3025)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3166..3188
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3182..3373)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5728..5747
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5767..5844)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
       61 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc
      121 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
      181 aaataatttt gtttaacttt aagaaggaga tataccatgg gccctgagcc ggctaaatca
      241 gcgccagctc ctaagaaagg atctaaaaag gccgtcacaa aagcgcagaa gaaggacggc
      301 aaaaaacgta aacgttcccg caaagaatca tatagcattt atgtctataa agttctcaaa
      361 caagtgcacc cagatactgg catttctagt aaggcgatgg gtatcatgaa ttcatttgta
      421 aacgacattt ttgagcgcat tgccggcgaa gcctcccgtc tcgctcacta taataagcgt
      481 tcaacaatca cctcacgtga gattcaaact gcggtccggt tgctcttgcc aggcgaatta
      541 gctaaacatg cagtgtctga gggcactaaa gcggtgacaa aatacacttc ggcgaaatga
      601 taaggatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
      661 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
      721 aggaggaact atatccggat atcccgcaag aggcccggca gtaccggcat aaccaagcct
      781 atgcctacag catccagggt gacggtgccg aggatgacga tgagcgcatt gttagatttc
      841 atacacggtg cctgactgcg ttagcaattt aactgtgata aactaccgca ttaaagctta
      901 tcgatgataa gctgtcaaac atgagaattc ttgaagacga aagggcctcg tgatacgcct
      961 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     1021 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1081 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     1141 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     1201 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     1261 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     1321 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt
     1381 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     1441 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     1501 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     1561 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     1621 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc
     1681 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     1741 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     1801 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     1861 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     1921 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     1981 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2041 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     2101 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     2161 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     2221 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     2281 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     2341 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     2401 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     2461 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     2521 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     2581 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     2641 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     2701 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     2761 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     2821 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     2881 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     2941 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca atggtgcact
     3001 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
     3061 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     3121 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     3181 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag
     3241 cgtggtcgtg aagcgattca cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt
     3301 tctccagaag cgttaatgtc tggcttctga taaagcgggc catgttaagg gcggtttttt
     3361 cctgtttggt cactgatgcc tccgtgtaag ggggatttct gttcatgggg gtaatgatac
     3421 cgatgaaacg agagaggatg ctcacgatac gggttactga tgatgaacat gcccggttac
     3481 tggaacgttg tgagggtaaa caactggcgg tatggatgcg gcgggaccag agaaaaatca
     3541 ctcagggtca atgccagcgc ttcgttaata cagatgtagg tgttccacag ggtagccagc
     3601 agcatcctgc gatgcagatc cggaacataa tggtgcaggg cgctgacttc cgcgtttcca
     3661 gactttacga aacacggaaa ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt
     3721 tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga ttcattctgc taaccagtaa
     3781 ggcaaccccg ccagcctagc cgggtcctca acgacaggag cacgatcatg cgcacccgtg
     3841 gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct gctggagatg gcggacgcga
     3901 tggatatgtt ctgccaaggg ttggtttgcg cattcacagt tctccgcaag aattgattgg
     3961 ctccaattct tggagtggtg aatccgttag cgaggtgccg ccggcttcca ttcaggtcga
     4021 ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg cagacaaggt atagggcggc
     4081 gcctacaatc catgccaacc cgttccatgt gctcgccgag gcggcataaa tcgccgtgac
     4141 gatcagcggt ccaatgatcg aagttaggct ggtaagagcc gcgagcgatc cttgaagctg
     4201 tccctgatgg tcgtcatcta cctgcctgga cagcatggcc tgcaacgcgg gcatcccgat
     4261 gccgccggaa gcgagaagaa tcataatggg gaaggccatc cagcctcgcg tcgcgaacgc
     4321 cagcaagacg tagcccagcg cgtcggccgc catgccggcg ataatggcct gcttctcgcc
     4381 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     4441 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     4501 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     4561 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     4621 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     4681 gcgctcactg cccgctttcc agtcgggaaa ctgtcgtgcc agctgcatta atgaatcggc
     4741 caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag
     4801 tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg
     4861 gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat
     4921 ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg caccaacgcg
     4981 cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag
     5041 catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat
     5101 ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt
     5161 atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc
     5221 gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg
     5281 ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac
     5341 attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat
     5401 cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc
     5461 gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat
     5521 cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag
     5581 caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc
     5641 catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac
     5701 gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg
     5761 tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa
     5821 ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca
     5881 ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca aggaatggtg
     5941 catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc
     6001 cgaaacaagc gctcatgagc ccg
//
