LOCUS       sequence-292455-        7612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human PHF19 in insect cells, , under a C3-cleavable 
            (PreScission Protease) N-terminal hexahistidine-MBP tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7612)
  AUTHORS   Zhang Q, McKenzie NJ, Warneford-Thomson R, Gail EH, Flanigan SF, 
            Owen BM, Lauman R, Levina V, Garcia BA, Schittenhelm RB, Bonasio R, 
            Davidovich C
  TITLE     RNA exploits an exposed regulatory site to inhibit the enzymatic 
            activity of PRC2.
  JOURNAL   Nat Struct Mol Biol. 2019 Mar;26(3):237-247. doi: 
            10.1038/s41594-019-0197-y. Epub 2019 Mar 4.
  PUBMED    30833789
REFERENCE   2  (bases 1 to 7612)
  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: 
            "10.1038/s41594-019-0197-y"; journalName: "Nat Struct Mol Biol"; 
            date: "2019-03"; volume: "26"; issue: "3"; pages: "237-247"
COMMENT     Sequence Label: pFB1.HMBP.PrS.PHF19
FEATURES             Location/Qualifiers
     source          1..7612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        171..262
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     187..204
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             323..340
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             347..1444
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1301..1310
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1418..1441
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1481..1504
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    3401..3535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3438..3457)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3492..3511
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3564..3729
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3913..4367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4000..4019)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4209..4230
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4393..4497
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4498..4566,4567..5358)
                     /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(4716..4735)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5529..6117
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6018..6037
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6260..6277
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6411..6635
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6702..7235)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7405..7410,7411..7427,7428..7433))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 cctggcttca ggagatcgga agacctcggc cgtcgcggcg cttgccggtg gtgctgaccc
       61 cggatgaagt ggttcgcatc ctcggttttc tggaaggcga gcatcgtttg ttcgcccagg
      121 actctagcta tagttctagt ggttggctac atactccgga atattaatag atcatggaga
      181 taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc gtaacagttt
      241 tgtaataaaa aaacctataa atattccgga ttattcatac cgtcccacca tcgggcgcgg
      301 atctcggtcc gaaaccatgg gtcatcacca tcatcatcac ggtaccaaaa ctgaagaagg
      361 taaactggta atctggatta acggcgataa aggctataac ggtctcgctg aagtcggtaa
      421 gaaattcgag aaagataccg gaattaaagt caccgttgag catccggata aactggaaga
      481 gaaattccca caggttgcgg caactggcga tggccctgac attatcttct gggcacacga
      541 ccgctttggt ggctacgctc aatctggcct gttggctgaa atcaccccgg acaaagcgtt
      601 ccaggacaag ctgtatccgt ttacctggga tgccgtacgt tacaacggca agctgattgc
      661 ttacccgatc gctgttgaag cgttatcgct gatttataac aaagatctgc tgccgaaccc
      721 gccaaaaacc tgggaagaga tcccggcgct ggataaagaa ctgaaagcga aaggtaagag
      781 cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg ccgctgattg ctgctgacgg
      841 gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa
      901 cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg attaaaaaca aacacatgaa
      961 tgcagacacc gattactcca tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac
     1021 catcaacggc ccgtgggcat ggtccaacat cgacaccagc aaagtgaatt atggtgtaac
     1081 ggtactgccg accttcaagg gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg
     1141 tattaacgcc gccagtccga acaaagagct ggcaaaagag ttcctcgaaa actatctgct
     1201 gactgatgaa ggtctggaag cggttaataa agacaaaccg ctgggtgccg tagcgctgaa
     1261 gtcttacgag gaagagttgg cgaaagatcc acgtattgcc gccaccatgg aaaacgccca
     1321 gaaaggtgaa atcatgccga acatcccgca gatgtccgct ttctggtatg ccgtgcgtac
     1381 tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat gaagccctga aagacgcgca
     1441 gactgcggcc gcaggcaccg attacgatat cccaacgacc ctggaagttc tgttccaggg
     1501 gcccggggag aatcgagctc tggatccagg gactcgggac tcctatggtg ccaccagcca
     1561 cctccccaac aagggggccc tggcgaaggt caagaacaac ttcaaagact tgatgtccaa
     1621 actgacggag ggccagtatg tgctgtgccg gtggacagat ggcctgtact acctcgggaa
     1681 gatcaagagg gtcagcagct ctaagcaaag ctgcctcgtg actttcgaag ataattccaa
     1741 atactgggtc ctatggaagg acatacagca tgccggtgtt ccaggagagg agcccaagtg
     1801 caacatctgc ctagggaaga catcagggcc gctgaatgag atcctcatct gcgggaagtg
     1861 tggcctgggt taccaccagc agtgccacat ccccatagcg ggcagtgctg accagcccct
     1921 gctcacacct tggttctgcc gacgctgcat cttcgcactg gctgtgcgga aaggcggcgc
     1981 gctgaagaag ggcgccatcg ccaggacgct gcaggccgtg aagatggtgc tgtcctacca
     2041 gcccgaggag ctcgagtggg actcgcccca tcgcaccaac cagcagcaat gctactgcta
     2101 ctgcggcggg cccggagaat ggtacctgcg gatgctgcaa tgttaccggt gcaggcagtg
     2161 gttccacgag gcctgcaccc agtgcctcaa tgagcccatg atgtttggtg accggtttta
     2221 cctgttcttc tgctccgtgt gtaaccaggg cccagagtac atcgagaggc tgcccctgcg
     2281 atgggtggat gtggttcacc tggccctcta taatctgggg gtacagagca agaagaagta
     2341 ctttgacttt gaggagattc tggcctttgt caaccaccac tgggagctcc tgcagcttgg
     2401 caagctcacc agcaccccag tgacagatcg aggaccacat ctcctcaacg ctctgaacag
     2461 ttataaaagc cggttcctct gcggcaagga gatcaagaag aagaagtgca tcttccgcct
     2521 gcgcatccgc gtcccaccca acccgccagg gaagctgctg cctgacaaag gactgctgcc
     2581 aaatgagaac agcgcctcct ctgagctgcg taagagagga aagagcaagc ctggtttgtt
     2641 gcctcacgaa ttccagcagc agaaaaggcg agtttataga agaaaaagat caaagttttt
     2701 gctggaagat gctattccca gtagtgactt cacctcagcc tggagcacca accaccacct
     2761 ggctagcata tttgacttca cgctggatga aattcaaagt ttaaaaagtg ccagctcagg
     2821 ccagaccttc ttctcagatg tcgactccac cgacgctgcc agcacctctg gctctgcctc
     2881 caccagcctc tcctatgact ccagatggac agtgggcagc cgaaagagga agctggcagc
     2941 caaggcatac atgcccctgc gggcaaagcg gtgggcagct gagctggatg gacgctgccc
     3001 ctcggacagc agtgcagagg gggcttcagt ccccgagcgg ccagacgaag gcattgacag
     3061 ccacacattt gagagcatca gtgaagatga ctcatccctg tcccacctca agtcatctat
     3121 caccaactac tttggtgcag ctgggcggtt ggcctgtggg gagaagtacc aggtgttggc
     3181 tcggagggtc acacctgagg gcaaggttca gtacctggtg gagtgggaag ggaccacccc
     3241 ttactgataa tagctcgagg catgcggtac caagcttgtc gagaagtact agaggatcat
     3301 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3361 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3421 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3481 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgatc
     3541 actgatatcg cctaggagat ccgaaccaga taagtgaaat ctagttccaa actattttgt
     3601 catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact
     3661 cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca actattttgt
     3721 ccgcccacag cggggcattt ttcttcctgt tatgttttta atcaaacatc ctgccaactc
     3781 catgtgacaa accgtcatct tcggctactt tttctctgtc acagaatgaa aatttttctg
     3841 tcatctcttc gttattaatg tttgtaattg actgaatatc aacgcttatt tgcagcctga
     3901 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3961 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4021 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4081 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4141 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4201 tcttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4261 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4321 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     4381 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4441 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4501 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4561 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4621 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4681 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4741 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4801 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4861 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4921 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4981 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5041 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5101 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5161 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5221 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5281 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5341 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5401 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5461 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5521 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5581 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5641 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     5701 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5761 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5821 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5881 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5941 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6001 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6061 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6121 gccagcaacg cggccttttt acggttcctg gccttttgct cacatgttct ttcctgcgtt
     6181 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6241 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     6301 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     6361 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     6421 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     6481 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     6541 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     6601 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     6661 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     6721 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     6781 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     6841 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     6901 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     6961 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     7021 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     7081 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     7141 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     7201 ccctgctgcg taacatcgtt gctgctgcgt aacatcaaac atcgacccac ggcgtaacgc
     7261 gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata acaagccatg
     7321 aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga ccagttgcgt
     7381 gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt caactgggtt
     7441 cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag cagcgaagtc
     7501 gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac gcatcgtcag
     7561 gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct gc
//
