LOCUS       sequence-360072-        6835 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express PC-H6 in E. coli. Purified PC-H6 was used for for generating
            antibodies in rabbits and for GST-CBP pull-down assays..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6835)
  AUTHORS   Tie F, Banerjee R, Fu C, Stratton CA, Fang M, Harte PJ
  TITLE     Polycomb inhibits histone acetylation by CBP by binding directly to 
            its catalytic domain.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Feb 9;113(6):E744-53. doi: 
            10.1073/pnas.1515465113. Epub 2016 Jan 22.
  PUBMED    26802126
REFERENCE   2  (bases 1 to 6835)
  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: 
            "10.1073/pnas.1515465113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-02-9- 9"; volume: "113"; issue: "6"; pages: "E744-53"
COMMENT     Sequence Label: pET11-PC-H6
FEATURES             Location/Qualifiers
     source          1..6835
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..15,16..32,33..38)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(213..260)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     256..274
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(331..348)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1021..1044)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             complement(1093..1122)
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     RBS             1529..1551
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1566..1590
                     /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)."
     primer_bind     complement(1590..1609)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1591..1609)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1693..1712)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        1918..1995
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1996..3078
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(2015..2034)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4390..4581
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4575..4597)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4683..4822
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4738..4757
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4837..4854)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5008..5596)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5088..5107)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5767..6558,6559..6627))
                     /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     6390..6409
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6628..6732)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6800..6818
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttctcatgtt tgacagctta tcatcgataa gctttaatgc ggtagtttat cacagttaaa
       61 ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat gcgctcatcg tcatcctcgg
      121 caccgtcacc ctggatgctg taggcatagg cttggttatg ccggtactgc cgggcctctt
      181 gcgggatatc cggatatagt tcctcctttc agcaaaaaac ccctcaagac ccgtttagag
      241 gccccaaggg gttatgctag ttattgctca gcggtggcag cagccaactc agcttccttt
      301 cgggctttgt tagcagccgg atccttatta atggtggtga tggtgatgca tagctactgg
      361 cgacgaatcg cctttcatgt cgcgctcacg aaaaaacccg cgctccgtct tgcactcgcg
      421 tatggtaacg gtctccaggt ttacggtgac gtctgtgatc accacccggt tggatatgtt
      481 gcatttggcc ggcagccaga agcgcggcgg aagggcacgc ggcgaaagag gagtaagcgg
      541 ctgtttctgg ttgatagcca ggttgttgca cggttttggt atgttattgt tctcggaagg
      601 aacctgtggc gcctgttggt tgccttcctc gttggctact tcttcttcct cagctccgga
      661 ctctgcgggc gttgtgttta tggcttctgt ggccagtggg gtggcctgct gctcagattt
      721 cagctgcgtt gcagcctcgc tggcaatctt ttccgcttgc tgctgatcct ggaagggctg
      781 ctgctggctg ggagttacct gctgggtcgg ctgaggctta atagtggggc catctgggga
      841 ggtcttaatc gtaactccaa tcttgcccga ttccttgagc acctcggctt tgcgtttggt
      901 gccgatcagc ggctgatcct cggagctgga ggagttgctg tccggctcgg ggacaaatga
      961 gttgtgcgtg aagctgctgt tcgaggagga gctgtgatca atgcgctgac gcttggactc
     1021 gtggtggtgg tgatgatgat ggtgggtcag cggagattcc gagcgccgtc cactgttgcg
     1081 ttcggacttg atgtggtgat gatggtggtg gtggtgatgg tgcttcttct ccttcttcga
     1141 ctccttatcg tgggtagccg atgaggtggt ggcttgatgc gtgtccacat cgttttcggt
     1201 gaaggtgtac tcatcctcct cggattccgg ctccggatcg ggttctttct ccttcttctt
     1261 aatgcctcgc ttggagggag ttccggagga tttgttcgtt tgttcgtaga tgtcgatgag
     1321 gcggcgatcc aggatgttta cctccggttc ccaggtgttg tagcgctggt tccagccctt
     1381 ccacttgaca cggtactcca cgacgccctt cttaacgcgc ttttggatga ttttctcagc
     1441 cgcgtacact agatcgactg gatcgtcggt cgcattgtcg cgccccaact tccccttact
     1501 ccccttgcct cgaccagtca tatgtatatc tccttcttaa agttaaacaa aattatttct
     1561 agaggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa tttcgcggga
     1621 tcgagatctc gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
     1681 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
     1741 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
     1801 tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc
     1861 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga gatcccggac
     1921 accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
     1981 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
     2041 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
     2101 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
     2161 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
     2221 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
     2281 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     2341 gcgcaacgcg tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     2401 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     2461 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     2521 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
     2581 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
     2641 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
     2701 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
     2761 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat
     2821 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg
     2881 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
     2941 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa
     3001 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     3061 ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc
     3121 gggatctcga ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg
     3181 gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca
     3241 ggtgccggca gcgctctggg tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat
     3301 gatcggcctg tcgcttgcgg tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac
     3361 tggtcccgcc accaaacgtt tcggcgagaa gcaggccatt atcgccggca tggcggccga
     3421 cgcgctgggc tacgtcttgc tggcgttcgc gacgcgaggc tggatggcct tccccattat
     3481 gattcttctc gcttccggcg gcatcgggat gcccgcgttg caggccatgc tgtccaggca
     3541 ggtagatgac gaccatcagg gacagcttca aggatcgctc gcggctctta ccagcctaac
     3601 ttcgatcatt ggaccgctga tcgtcacggc gatttatgcc gcctcggcga gcacatggaa
     3661 cgggttggca tggattgtag gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg
     3721 cggtgcatgg agccgggcca cctcgacctg aatggaagcc ggcggcacct cgctaacgga
     3781 ttcaccactc caagaattgg agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc
     3841 aacccttggc agaacatatc catcgcgtcc gccatctcca gcagccgcac gcggcgcatc
     3901 tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc
     3961 cggctaggct ggcggggttg ccttactggt tagcagaatg aatcaccgat acgcgagcga
     4021 acgtgaagcg actgctgctg caaaacgtct gcgacctgag caacaacatg aatggtcttc
     4081 ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag cgccctgcac cattatgttc
     4141 cggatctgca tcgcaggatg ctgctggcta ccctgtggaa cacctacatc tgtattaacg
     4201 aagcgctggc attgaccctg agtgattttt ctctggtccc gccgcatcca taccgccagt
     4261 tgtttaccct cacaacgttc cagtaaccgg gcatgttcat catcagtaac ccgtatcgtg
     4321 agcatcctct ctcgtttcat cggtatcatt acccccatga acagaaatcc cccttacacg
     4381 gaggcatcag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt tatcagaagc
     4441 cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca ggcagacatc
     4501 tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc gcgtttcggt
     4561 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     4621 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     4681 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     4741 catcagagca gattgtactg agagtgcacc attgcggtgt gaaataccgc acagatgcgt
     4801 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4861 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4921 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4981 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5041 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5101 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5161 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5221 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5281 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5341 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5401 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5461 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5521 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5581 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5641 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5701 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5761 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5821 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5881 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5941 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6001 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6061 gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6121 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6181 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6241 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6301 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6361 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     6421 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6481 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6541 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6601 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6661 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6721 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6781 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaa
//
