LOCUS       sequence-445645-        7932 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of p300 HAT domain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7932)
  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 7932)
  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: pETduet+p300HAT
FEATURES             Location/Qualifiers
     source          1..7932
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        106..124
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    125..149
                     /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             164..186
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             199..225
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     CDS             226..246
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     regulatory      1147..1156
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        1464..1482
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1483..1507
                     /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)."
     CDS             3003..3047
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3085..3103)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3099..3146
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3183..3638
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3270..3289)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3480..3501
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3753..4544,4545..4613))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4376..4395
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4614..4706)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4787..5375
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5276..5295
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5529..5546
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5561..5700
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5626..5645)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5786..5808
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5802..5993)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6565..7647)
                     /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     7609..7628
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7648..7725)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(7931..7932,1..18)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
       61 tgcgtccggc gtagaggatc gagatcgatc tcgatcccgc gaaattaata cgactcacta
      121 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
      181 aggagatata ccatgggaca tcaccatcat caccaccatc atcacgaaaa cctgtatttt
      241 cagggatcca agttttctgc taaaaggttg ccatctacca gacttggcac ctttctagag
      301 aatcgtgtga atgactttct gaggcgacag aatcaccctg agtcaggaga ggtcactgtt
      361 agagtagttc atgcttctga caaaaccgtg gaagtaaaac caggcatgaa agcaaggttt
      421 gtggacagtg gagagatggc agaatccttt ccataccgaa ccaaagccct ctttgccttt
      481 gaagaaattg atggtgttga cctgtgcttc tttggcatgc atgttcaaga gtatggctct
      541 gactgccctc cacccaacca gaggagagta tacatatctt acctcgatag tgttcatttc
      601 ttccgtccta aatgcttgag gactgcagtc tatcatgaaa tcctaattgg atatttagaa
      661 tatgtcaaga aattaggtta cacaacaggg catatttggg catgtccacc aagtgaggga
      721 gatgattata tcttccattg ccatcctcct gaccagaaga tacccaagcc caagcgactg
      781 caggaatggt acaaaaaaat gcttgacaag gctgtatcag agcgtattgt ccatgactac
      841 aaggatattt ttaaacaagc tactgaagat agattaacaa gtgcaaagga attgccttat
      901 ttcgagggtg atttctggcc caatgttctg gaagaaagca ttaaggaact ggaacaggag
      961 gaagaagaga gaaaacgaga ggaaaacacc agcaatgaaa gcacagatgt gaccaaggga
     1021 gacagcaaaa atgctaaaaa gaagaataat aagaaaacca gcaaaaataa gagcagcctg
     1081 agtaggggca acaagaagaa acccgggatg cccaatgtat ctaacgacct ctcacagaaa
     1141 ctatatgcca ccatggagaa gcataaagag gtcttctttg tgatccgcct cattgctggc
     1201 cctgctgcca actccctgcc tcccattgtt gatcctgatc ctctcatccc ctgcgatctg
     1261 atggatggtc gggatgcgtt tctcacgctg gcaagggaca agcacctgga gttctcttca
     1321 ctccgaagag cccagtggtc caccatgtgc atgctggtgg agctgcacac gcagagccag
     1381 gacggctcga gctgataagc ggccgcataa tgcttaagtc gaacagaaag taatcgtatt
     1441 gtacacggcc gcataatcga aattaatacg actcactata ggggaattgt gagcggataa
     1501 caattcccca tcttagtata ttagttaagt ataagaagga gatatacata gttcatatgg
     1561 accccttggc agtgtcagca gcttcagtag tgtcaatgtc caatgacgtt ttgaaaccag
     1621 agacgcccaa ggggccaatc ataatcagta aaaacccatc aaatggtatt ttctatggtc
     1681 cctccttcac taaacgagag tctctcaatg ctcgaatgtt tctgaaatac tatggtgcac
     1741 acaaattttt agacacttac ctccccgagg atttgaactc gttatacatt tactatctta
     1801 tcaagttgct aggctttgaa gttaaagatc aagcgcttat cggcaccatc aacagtattg
     1861 tccatatcaa ctcgcaagag cgtgttcaag atttgggaag tgcaatatct gtcacaaatg
     1921 ttgaagaccc attggcaaaa aagcaaacag ttcgtctaat caaagatttg caaagagcaa
     1981 ttaacaaagt tctatgtaca agattaagat tatccaattt tttcactatt gatcatttta
     2041 ttcgaaaatt acataccgct agaaaaattt tggtcctgac tggtgcaggt gtttcaactt
     2101 cattaggcat cccggacttc agatcttctg aggggttcta ttcaaagatc aaacatttgg
     2161 ggctcgatga tccccaagac gttttcaatt acaatatatt tatgcacgac ccctctgttt
     2221 tctataatat tgccaatatg gttttacctc cagaaaaaat ttattctcca ttgcatagtt
     2281 tcattaagat gctacaaatg aaagggaaat tattgagaaa ttatactcaa aacattgata
     2341 atttggaatc ttatgcggga ataagcacag ataaactggt gcagtgccat ggctcttttg
     2401 ctactgccac ctgcgttacc tgccattgga acctacccgg tgagaggata tttaataaaa
     2461 ttagaaacct cgaacttcca ctatgcccgt actgttacaa aaaaagaaga gaatatttcc
     2521 cagagggata taataataaa gtaggtgttg ctgcatcaca gggttcaatg tcggaaaggc
     2581 ctccatatat ccttaactca tatggcgttc tcaaaccaga tatcacattc tttggcgaag
     2641 cactgccaaa taaatttcat aagagcattc gcgaagatat cttagaatgt gatttgttga
     2701 tttgcattgg gacaagttta aaagtagcgc cagtgtctga aatcgtaaac atggttcctt
     2761 cccacgttcc ccaagtcctg attaatcgtg atcccgtcaa gcacgcagaa tttgatttat
     2821 ctcttttggg gtactgtgat gacattgcag ctatggtagc ccaaaaatgt ggctggacga
     2881 ttccgcataa gaaatggaac gatttgaaga acaagaactt taaatgccaa gagaaggata
     2941 agggcgtgta tgtcgttaca tcagatgaac atcccaaaac cctctaatga ctcgagtctg
     3001 gtaaagaaac cgctgctgcg aaatttgaac gccagcacat ggactcgtct actagcgcag
     3061 cttaattaac ctaggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     3121 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
     3181 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     3241 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     3301 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     3361 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     3421 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     3481 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     3541 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     3601 taacgcgaat tttaacaaaa tattaacgtt tacaatttct ggcggcacga tggcatgaga
     3661 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3721 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3781 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3841 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3901 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3961 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4021 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4081 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4141 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4201 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4261 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4321 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4381 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4441 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4501 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4561 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4621 ctttttcaat catgattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4681 tgaatgtatt tagaaaaata aacaaatagg tcatgaccaa aatcccttaa cgtgagtttt
     4741 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4801 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4861 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4921 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4981 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5041 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5101 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5161 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5221 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5281 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5341 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5401 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5461 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5521 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     5581 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     5641 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     5701 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5761 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5821 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     5881 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     5941 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     6001 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     6061 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     6121 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     6181 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     6241 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     6301 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     6361 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     6421 cgggtcctca acgacaggag cacgatcatg ctagtcatgc cccgcgccca ccggaaggag
     6481 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     6541 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6601 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     6661 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     6721 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     6781 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     6841 cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     6901 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     6961 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     7021 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     7081 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     7141 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     7201 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     7261 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     7321 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     7381 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     7441 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     7501 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     7561 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     7621 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     7681 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     7741 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     7801 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     7861 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     7921 cttccccatc gg
//
