LOCUS       sequence-360059-        6597 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A PCR product encoding GST-H3 (Drosophila histone H3) was was 
            inserted into a modified pAct-5c vector by Sac I and Sal I sites. 
            Expression in S2 cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6597)
  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 6597)
  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
FEATURES             Location/Qualifiers
     source          1..6597
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        322..2831
                     /label=Ac5 promoter
                     /note="Drosophila melanogaster actin 5C promoter"
     CDS             2869..3522
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3480..3502
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3529..3546
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     intron          4107..4172
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4302..4322
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    4747..4881
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4784..4803)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4838..4857
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             join(4978..5046,5047..5838)
                     /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(5196..5215)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6009..6597
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6498..6517
                     /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 taaaaaaaaa tcatgaatgg catcaactct gaatcaaatc tttgcagatg
      361 cacctacttc tcatttccac tgtcacatca tttttccaga tctcgctgcc tgttatgtgg
      421 cccacaaacc aagacacgtt ttatggccat taaagctggc tgatcgtcgc caaacaccaa
      481 atacataatg aatatgtaca cattcgagaa agaagcgatc aaagaagcgt cttcgggcgg
      541 agtaggagaa tgcggaggag aaggagaacg agctgatcta gtatctctcc acaatccaat
      601 gccaactgac caactggcca tattcggagc aatttgaagc caatttccat cgcctggcga
      661 tcgctccatt cttggctata tgtttttcac cgttacccgg ggccattttc aaagactcgt
      721 cggcaagata agattgtgtc actcgctgtc tctcttcatt tgtcgaagaa tgctgaggaa
      781 tttcgcgatg acgtcggcga gtattttgaa gaatgagaat aatttgtatt tatacgaaaa
      841 tcagttagtg gaattttcta caaaaacatg ttatctatag ataattttgt tgcaaaatat
      901 gttgactatg acaaagattg tatgtatata cctttaatgt attctcattt tcttatgtat
      961 ttataatggc aatgatgata ctgatgatat tttaagatga tgccagacca aaaggcttga
     1021 atttctgcgt cttttgccga acgcagtgca tgtgcaattg ttgttttttg gaatattcaa
     1081 ttttcggact gtccgctttg atttcagttt cttggcttat tcaaaaagca aagtaaagcc
     1141 aaaaaagcga gatggcaata ccaaatgcgg caaaacggta gtggaaggaa aggggtgcgg
     1201 ggcagcggaa ggaagggtgg ggcggggcgt ggcggggtct gtggctgggc gcgacgtcac
     1261 cgacgttgga gccactcctt tgaccatgtg tgcgtgtgtg tattattcgt gtctcgccac
     1321 tcgccggttg tttttttctt tttatgctgc gctctctcta gcgccatctc gcttacgcat
     1381 gctcaacgca ccgcatgttg ccgtttcctt ttatgcgtca ttttggctcg aaataggcaa
     1441 ttatttaaac aaagattagt caacgaaaac gctaaaataa ataagtctac aatatggtta
     1501 cttattgcca tgtgtgtgca gccaacgata gcaacaaaag caacaacaca ggtggctttc
     1561 cctctttcac tttttgtttg caagccgcgt gcgagcaaga cggcacgacc ggcaaacgca
     1621 attacgctga caaagagcag acgaagtttt ggcgaaaaac atcaaggcgc ctgatacgaa
     1681 tgcatttgca ataacaattg cgatatttaa tattgtttat gaagctgttt gacttcaaaa
     1741 cacacaaaaa aaaaaataaa acaaattatt tgaaagagaa ttaggaatcg gacgcttatc
     1801 gttagggtaa caacaagaaa tgcttactga gtcacagcct ctggaaaact gccgcaagcc
     1861 agagagagag agaaaaagag ggagagcagc ttagaccgca tgtgcttgtg tgtgaggcgt
     1921 ctctctcttc gtctctgttg cgcaaacgca tagactgcac tgaaaaaatc gattacctat
     1981 tttttatgaa tgaatatttg cactattact attcaaaact attaagatag caatcacatt
     2041 caatagccaa atactatacc acctgagcga tgcaacgaaa tgatcaattt gagcaaaaat
     2101 gctgcatatt taggacggca tcattataga aatgcttctt gctgtgtact tttctctcgt
     2161 ctggcagctg tttcgccgtt attgttaaaa ccggcttaag ttaggtgtgt tttctacgac
     2221 tagtgaatgc cctactagaa gatgtgtgtt gcacaaaatg tccctggaat aaccaatttg
     2281 aagtgcagat agcagtaaac gtaagctaat atgaatatta tttaactgta atgttttaat
     2341 atcgctggac attactaata aacccactat aaacacatgt acatatgtat gttttggcat
     2401 acaatgagta gttggggaaa aaatgtgtaa aagcaccgtg accatcacag cataaagata
     2461 accagctgaa gtatcgaata tgagtaaccc ccaaattgaa tcacatgccg caactgatag
     2521 gacccatgga agtacactct tcatggcgat atacaagaca cacacaagca cgaacaccca
     2581 gttgcggagg aaattctccg taaatgaaaa cccaatcggc gaacaattca tacccatata
     2641 tggtaaaagt tttgaacgcg acttgagagc ggagagcatt gcggctgata aggttttagc
     2701 gctaagcggg ctttataaaa cgggctgcgg gaccagtttt catatcacta ccgtttgagt
     2761 tcttgtgctg tgtggatact cctcccgaca caaagccgct ccatcagcca gcagtcgtct
     2821 aatccagaga ccccggatcc agatatcgag gcctgtctag aagagctcat gtcccctata
     2881 ctaggttatt ggaaaattaa gggccttgtg caacccactc gacttctttt ggaatatctt
     2941 gaagaaaaat atgaagagca tttgtatgag cgcgatgaag gtgataaatg gcgaaacaaa
     3001 aagtttgaat tgggtttgga gtttcccaat cttccttatt atattgatgg tgatgttaaa
     3061 ttaacacagt ctatggccat catacgttat atagctgaca agcacaacat gttgggtggt
     3121 tgtccaaaag agcgtgcaga gatttcaatg cttgaaggag cggttttgga tattagatac
     3181 ggtgtttcga gaattgcata tagtaaagac tttgaaactc tcaaagttga tttccttagc
     3241 aagctacctg aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac atatttaaat
     3301 ggtgatcatg taacccatcc tgacttcatg ttgtatgacg ctcttgatgt tgttttatac
     3361 atggacccaa tgtgcctgga tgcgttccca aaattagttt gttttaaaaa acgtattgaa
     3421 gctatcccac aaattgataa gtacttgaaa tccagcaagt atatagcatg gcctttgcag
     3481 ggctggcaag ccacgtttgg tggtggcgac catcctccaa aatcggatct ggttccgcgt
     3541 ggatcccata tggctcgtac caagcaaact gctcgcaaat cgactggtgg aaaggcgcca
     3601 cgcaaacaac tggctactaa ggccgctcgc aagagtgctc cagccaccgg aggtgtgaag
     3661 aagccccacc gctatcgccc tggaaccgtg gccttgcgtg aaattcgtcg ctaccaaaag
     3721 agcaccgagc ttctaatccg caagctgcct ttccagcgtc tggtgcgtga aatcgctcag
     3781 gactttaaga cggacttgcg attccagagc tcggcggtta tggctctgca ggaagctagc
     3841 gaagcctacc tggttggtct cttcgaagat accaacttgt gtgccattca tgccaagcgt
     3901 gtcaccataa tgcccaaaga catccagtta gcgcgacgca ttcgcggcga gcgtgcttaa
     3961 gaattccgaa tcactagtga attcgcggcc gcctgcaggt cgacgccggc tgactgactg
     4021 actctagcag gatctttgtg aaggaacctt acttctgtgg tgtgacataa ttggacaaac
     4081 tacctacaga gatttaaagc tctaaggtaa atataaaatt tttaagtgta taatgtgtta
     4141 aactactgat tctaattgtt tgtgtatttt agattccaac ctatggaact gatgaatggg
     4201 agcagtggtg gaatgccttt aatgaggaaa acctgttttg ctcagaagaa atgccatcta
     4261 gtgatgatga ggctactgct gactctcaac attctactcc tccaaaaaag aagagaaagg
     4321 tagaagaccc aaaggacttt ccttcagaat tgctaagttt tttgagtcat gctgtgttta
     4381 gtaatagaac tcttgcttgc tttgctattt acaccacaaa ggaaaaagct gcactgctat
     4441 acaagaaaat tatggaaaaa tattctgtaa cctttataag taggcataac agttataatc
     4501 ataacatact gttttttctt actccacaca ggcatagagt gtctgctatt aataactatg
     4561 ctcaaaaatt gtgtaccttt agctttttaa tttgtaaagg ggttaataag gaatatttga
     4621 tgtatagtgc cttgactaga gatcataatc agccatacca catttgtaga ggttttactt
     4681 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt
     4741 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4801 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4861 gtatcttatc atgtctggat cccgtttaaa ctacgcgtaa ttcaaacagg gttctggcgt
     4921 cgttctcgta ctgttttccc caggccagtg ctttagcgtt attgaaaaag gaagagtatg
     4981 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5041 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5101 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5161 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5221 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5281 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5341 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5401 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5461 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5521 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5581 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5641 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5701 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5761 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5821 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5881 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5941 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6001 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6061 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6121 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6181 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6241 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6301 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6361 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6421 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6481 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6541 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaa
//
