LOCUS       sequence-360061-        6596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a K27Q point mutation (AAG/CAG) of Drosophila histone H3 was 
            generated from pAct-GST-H3. Expression in S2 cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6596)
  AUTHORS   Tie F, Banerjee R, Stratton CA, Prasad-Sinha J, Stepanik V, Zlobin 
            A, Diaz MO, Scacheri PC, Harte PJ
  TITLE     CBP-mediated acetylation of histone H3 lysine 27 antagonizes 
            Drosophila Polycomb silencing.
  JOURNAL   Development. 2009 Sep;136(18):3131-41. doi: 10.1242/dev.037127.
  PUBMED    19700617
REFERENCE   2  (bases 1 to 6596)
  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: "10.1242/dev"; 
            journalName: "Development"; date: "2009-09"; volume: "136"; issue: 
            "18"; pages: "3131-41"
COMMENT     Sequence Label: pAct-GST-H3(K27Q)
FEATURES             Location/Qualifiers
     source          1..6596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        321..2830
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2868..3521
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3479..3501
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3528..3545
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     intron          4106..4171
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4301..4321
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4746..4880
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4783..4802)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4837..4856
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             join(4977..5045,5046..5837)
                     /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     complement(5195..5214)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6008..6596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6497..6516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcaggcaa
      241 ctcgtgaaag gtaggcggat cagcggccgc agcatgcaat tctatattct aaaaacacaa
      301 atgatacttc taaaaaaaat catgaatggc atcaactctg aatcaaatct ttgcagatgc
      361 acctacttct catttccact gtcacatcat ttttccagat ctcgctgcct gttatgtggc
      421 ccacaaacca agacacgttt tatggccatt aaagctggct gatcgtcgcc aaacaccaaa
      481 tacataatga atatgtacac attcgagaaa gaagcgatca aagaagcgtc ttcgggcgga
      541 gtaggagaat gcggaggaga aggagaacga gctgatctag tatctctcca caatccaatg
      601 ccaactgacc aactggccat attcggagca atttgaagcc aatttccatc gcctggcgat
      661 cgctccattc ttggctatat gtttttcacc gttacccggg gccattttca aagactcgtc
      721 ggcaagataa gattgtgtca ctcgctgtct ctcttcattt gtcgaagaat gctgaggaat
      781 ttcgcgatga cgtcggcgag tattttgaag aatgagaata atttgtattt atacgaaaat
      841 cagttagtgg aattttctac aaaaacatgt tatctataga taattttgtt gcaaaatatg
      901 ttgactatga caaagattgt atgtatatac ctttaatgta ttctcatttt cttatgtatt
      961 tataatggca atgatgatac tgatgatatt ttaagatgat gccagaccaa aaggcttgaa
     1021 tttctgcgtc ttttgccgaa cgcagtgcat gtgcaattgt tgttttttgg aatattcaat
     1081 tttcggactg tccgctttga tttcagtttc ttggcttatt caaaaagcaa agtaaagcca
     1141 aaaaagcgag atggcaatac caaatgcggc aaaacggtag tggaaggaaa ggggtgcggg
     1201 gcagcggaag gaagggtggg gcggggcgtg gcggggtctg tggctgggcg cgacgtcacc
     1261 gacgttggag ccactccttt gaccatgtgt gcgtgtgtgt attattcgtg tctcgccact
     1321 cgccggttgt ttttttcttt ttatgctgcg ctctctctag cgccatctcg cttacgcatg
     1381 ctcaacgcac cgcatgttgc cgtttccttt tatgcgtcat tttggctcga aataggcaat
     1441 tatttaaaca aagattagtc aacgaaaacg ctaaaataaa taagtctaca atatggttac
     1501 ttattgccat gtgtgtgcag ccaacgatag caacaaaagc aacaacacag gtggctttcc
     1561 ctctttcact ttttgtttgc aagccgcgtg cgagcaagac ggcacgaccg gcaaacgcaa
     1621 ttacgctgac aaagagcaga cgaagttttg gcgaaaaaca tcaaggcgcc tgatacgaat
     1681 gcatttgcaa taacaattgc gatatttaat attgtttatg aagctgtttg acttcaaaac
     1741 acacaaaaaa aaaaataaaa caaattattt gaaagagaat taggaatcgg acgcttatcg
     1801 ttagggtaac aacaagaaat gcttactgag tcacagcctc tggaaaactg ccgcaagcca
     1861 gagagagaga gaaaaagagg gagagcagct tagaccgcat gtgcttgtgt gtgaggcgtc
     1921 tctctcttcg tctctgttgc gcaaacgcat agactgcact gaaaaaatcg attacctatt
     1981 ttttatgaat gaatatttgc actattacta ttcaaaacta ttaagatagc aatcacattc
     2041 aatagccaaa tactatacca cctgagcgat gcaacgaaat gatcaatttg agcaaaaatg
     2101 ctgcatattt aggacggcat cattatagaa atgcttcttg ctgtgtactt ttctctcgtc
     2161 tggcagctgt ttcgccgtta ttgttaaaac cggcttaagt taggtgtgtt ttctacgact
     2221 agtgaatgcc ctactagaag atgtgtgttg cacaaaatgt ccctggaata accaatttga
     2281 agtgcagata gcagtaaacg taagctaata tgaatattat ttaactgtaa tgttttaata
     2341 tcgctggaca ttactaataa acccactata aacacatgta catatgtatg ttttggcata
     2401 caatgagtag ttggggaaaa aatgtgtaaa agcaccgtga ccatcacagc ataaagataa
     2461 ccagctgaag tatcgaatat gagtaacccc caaattgaat cacatgccgc aactgatagg
     2521 acccatggaa gtacactctt catggcgata tacaagacac acacaagcac gaacacccag
     2581 ttgcggagga aattctccgt aaatgaaaac ccaatcggcg aacaattcat acccatatat
     2641 ggtaaaagtt ttgaacgcga cttgagagcg gagagcattg cggctgataa ggttttagcg
     2701 ctaagcgggc tttataaaac gggctgcggg accagttttc atatcactac cgtttgagtt
     2761 cttgtgctgt gtggatactc ctcccgacac aaagccgctc catcagccag cagtcgtcta
     2821 atccagagac cccggatcca gatatcgagg cctgtctaga agagctcatg tcccctatac
     2881 taggttattg gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg
     2941 aagaaaaata tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa
     3001 agtttgaatt gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat
     3061 taacacagtc tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt
     3121 gtccaaaaga gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg
     3181 gtgtttcgag aattgcatat agtaaagact ttgaaactct caaagttgat ttccttagca
     3241 agctacctga aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg
     3301 gtgatcatgt aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca
     3361 tggacccaat gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag
     3421 ctatcccaca aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg
     3481 gctggcaagc cacgtttggt ggtggcgacc atcctccaaa atcggatctg gttccgcgtg
     3541 gatcccatat ggctcgtacc aagcaaactg ctcgcaaatc gactggtgga aaggcgccac
     3601 gcaaacaact ggctactaag gccgctcgcc agagtgctcc agccaccgga ggtgtgaaga
     3661 agccccaccg ctatcgccct ggaaccgtgg ccttgcgtga aattcgtcgc taccaaaaga
     3721 gcaccgagct tctaatccgc aagctgcctt tccagcgtct ggtgcgtgaa atcgctcagg
     3781 actttaagac ggacttgcga ttccagagct cggcggttat ggctctgcag gaagctagcg
     3841 aagcctacct ggttggtctc ttcgaagata ccaacttgtg tgccattcat gccaagcgtg
     3901 tcaccataat gcccaaagac atccagttag cgcgacgcat tcgcggcgag cgtgcttaag
     3961 aattccgaat cactagtgaa ttcgcggccg cctgcaggtc gacgccggct gactgactga
     4021 ctctagcagg atctttgtga aggaacctta cttctgtggt gtgacataat tggacaaact
     4081 acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat aatgtgttaa
     4141 actactgatt ctaattgttt gtgtatttta gattccaacc tatggaactg atgaatggga
     4201 gcagtggtgg aatgccttta atgaggaaaa cctgttttgc tcagaagaaa tgccatctag
     4261 tgatgatgag gctactgctg actctcaaca ttctactcct ccaaaaaaga agagaaaggt
     4321 agaagaccca aaggactttc cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag
     4381 taatagaact cttgcttgct ttgctattta caccacaaag gaaaaagctg cactgctata
     4441 caagaaaatt atggaaaaat attctgtaac ctttataagt aggcataaca gttataatca
     4501 taacatactg ttttttctta ctccacacag gcatagagtg tctgctatta ataactatgc
     4561 tcaaaaattg tgtaccttta gctttttaat ttgtaaaggg gttaataagg aatatttgat
     4621 gtatagtgcc ttgactagag atcataatca gccataccac atttgtagag gttttacttg
     4681 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     4741 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4801 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4861 tatcttatca tgtctggatc ccgtttaaac tacgcgtaat tcaaacaggg ttctggcgtc
     4921 gttctcgtac tgttttcccc aggccagtgc tttagcgtta ttgaaaaagg aagagtatga
     4981 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5041 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5101 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5161 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5221 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5281 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5341 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5401 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5461 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5521 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5581 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5641 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5701 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5761 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5821 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5881 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     5941 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6001 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6061 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6121 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaa
//
