LOCUS       sequence-307924-        6053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of cysteine-free human histone H3 (c111A).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6053)
  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 6053)
  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+H3
FEATURES             Location/Qualifiers
     source          1..6053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     65..84
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        65..83
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    84..108
                     /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             123..145
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(619..637)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      633..680
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(855..860,861..877,878..883))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(910..928)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        996..1100
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1101..1169,1170..1961)
                     /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(1319..1338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2132..2720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2621..2640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2874..2891
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2906..3045
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2971..2990)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3131..3153
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3147..3338)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5693..5712
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5732..5809)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6015..6034
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg
       61 aaattaatac gactcactat aggggaattg tgagcggata acaattcccc tctagaaata
      121 attttgttta actttaagaa ggagatatac catgggcgct cgcacgaagc aaacagcccg
      181 caagagtact ggtggaaaag cgcctcggaa gcagctggcg acgaaagctg cacggaagtc
      241 ggctccagcg acggggggcg tcaagaagcc acaccgttat cggccgggca ccgttgctct
      301 tcgtgaaatc cgtcgctacc agaaaagcac agagttgctt attcggaaac tcccatttca
      361 gcgcctggtt cgtgaaattg cccaagattt caagaccgat ctccggttcc aaagctcagc
      421 cgtgatggcg ctccaagagg cttcggaggc ctacttagta ggtttattcg aggacactaa
      481 cttagccgcg atccatgcta agcgtgtgac gattatgccg aaggatatcc aacttgcgcg
      541 tcggattcgt ggagagcggg cctgataagg atccggctgc taacaaagcc cgaaaggaag
      601 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      661 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatatccc gcaagaggcc
      721 cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg tgccgaggat
      781 gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc aatttaactg
      841 tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag aattcttgaa
      901 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
      961 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1021 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1081 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1141 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1201 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1261 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1321 tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac
     1381 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1441 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1501 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     1561 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     1621 acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg
     1681 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     1741 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     1801 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     1861 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1921 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1981 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2041 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2101 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2161 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2221 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     2281 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2341 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2401 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2461 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2521 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2581 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2641 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2701 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2761 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2821 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2881 cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg
     2941 gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     3001 ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca
     3061 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     3121 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     3181 aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt
     3241 tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag
     3301 cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga
     3361 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     3421 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     3481 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     3541 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     3601 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     3661 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     3721 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     3781 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     3841 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     3901 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     3961 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     4021 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     4081 ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt aggctggtaa
     4141 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
     4201 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
     4261 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     4321 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     4381 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     4441 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     4501 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     4561 aggagctgac tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag
     4621 tgagctaact tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaactgtc
     4681 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4741 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     4801 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     4861 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     4921 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     4981 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     5041 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     5101 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     5161 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     5221 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     5281 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     5341 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     5401 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     5461 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     5521 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     5581 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     5641 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     5701 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     5761 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     5821 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     5881 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     5941 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     6001 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgt
//
