LOCUS       sequence-292452-        7573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human PHF1 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 7573)
  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 7573)
  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.PHF1
FEATURES             Location/Qualifiers
     source          1..7573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        312..403
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     328..345
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             464..481
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             488..1585
                     /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      1442..1451
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1559..1582
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1622..1645
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    2093..2107
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     polyA_signal    3503..3637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3540..3559)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3594..3613
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3666..3831
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4015..4469
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4102..4121)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4311..4332
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4495..4599
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4600..4668,4669..5460)
                     /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(4818..4837)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5631..6219
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6120..6139
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6362..6379
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6513..6737
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6804..7337)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7507..7512,7513..7529,7530..7535))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 acccggcaac cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc
       61 gcaaggtttc ggtctccacg catcgtcagg cattggcggc cttgctgttc ttctacggca
      121 aggtgctgtg cacggatctg ccctggcttc aggagatcgg aagacctcgg ccgtcgcggc
      181 gcttgccggt ggtgctgacc ccggatgaag tggttcgcat cctcggtttt ctggaaggcg
      241 agcatcgttt gttcgcccag gactctagct atagttctag tggttggcta catactccgg
      301 aatattaata gatcatggag ataattaaaa tgataaccat ctcgcaaata aataagtatt
      361 ttactgtttt cgtaacagtt ttgtaataaa aaaacctata aatattccgg attattcata
      421 ccgtcccacc atcgggcgcg gatctcggtc cgaaaccatg ggtcatcacc atcatcatca
      481 cggtaccaaa actgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa
      541 cggtctcgct gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga
      601 gcatccggat aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga
      661 cattatcttc tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga
      721 aatcaccccg gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg
      781 ttacaacggc aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa
      841 caaagatctg ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga
      901 actgaaagcg aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg
      961 gccgctgatt gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat
     1021 taaagacgtg ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct
     1081 gattaaaaac aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa
     1141 taaaggcgaa acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag
     1201 caaagtgaat tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt
     1261 cgttggcgtg ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga
     1321 gttcctcgaa aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc
     1381 gctgggtgcc gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc
     1441 cgccaccatg gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc
     1501 tttctggtat gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga
     1561 tgaagccctg aaagacgcgc agactgcggc cgcaggcacc gattacgata tcccaacgac
     1621 cctggaagtt ctgttccagg ggcccggggc gcagcccccc cggctgagcc gctctggtgc
     1681 ctcctcactt tgggacccag cttctcctgc tcccacctct ggccccaggc ctcggctttg
     1741 ggagggtcaa gatgtgctgg ccagatggac tgatgggctg ctatacttgg gtaccatcaa
     1801 aaaggtggac agtgctaggg aggtgtgtct ggtccagttt gaggatgatt cgcagtttct
     1861 ggttctatgg aaagacatta gccctgctgc cctccctgga gaggaactcc tctgttgtgt
     1921 ctgtcgctct gagactgtgg tccctgggaa ccggctggtc agctgtgaga agtgtcgcca
     1981 tgcttatcac caggactgcc atgttcccag ggctccagcc cctggagagg gagagggcac
     2041 atcctgggta tgccgccagt gtgtctttgc gatcgccacc aagaggggag gtgccctgaa
     2101 gaagggcccc tatgcccggg ccatgctggg tatgaagctt tctctgccat atggactgaa
     2161 ggggctggac tgggatgctg gacatctgag caaccgacag cagagttact gttactgtgg
     2221 tggccctggg gagtggaacc tgaaaatgct gcagtgccgg agctgcctgc agtggttcca
     2281 tgaggcctgc acccagtgtc tgagcaagcc cctcctctat ggggacaggt tctatgaatt
     2341 tgaatgctgt gtgtgtcgcg ggggccctga gaaagtccgg agactacagc ttcgctgggt
     2401 ggatgtggcc catcttgtcc tgtatcacct cagtgtttgc tgtaagaaga aatactttga
     2461 ttttgatcgt gagatcctcc ccttcacttc tgagaattgg gacagtttgc tcctggggga
     2521 gctttcagac acccccaaag gagaacgttc ttccaggctc ctctctgctc ttaacagcca
     2581 caaggaccgt ttcatttcag ggagagagat taagaagagg aaatgtttgt ttggtctaca
     2641 tgctcggatg cctccccctg tggagccccc tactggagat ggagcactca ccagcttccc
     2701 ttcagggcag ggccctgggg gaggtgtctc acgtcccctg gggaagcgcc ggaggccgga
     2761 gccagagccc ctgaggagga ggcagaaggg gaaagtggag gagctggggc caccctcagc
     2821 agtgcgcaat cagcccgagc cccaggagca gagggagcgg gctcatctgc agagggcact
     2881 gcaggcctca gtgtctccac catcccccag ccctaaccag agttaccagg gcagcagcgg
     2941 ctacaacttc cggcccacag atgcccgctg cctgcccagc agccccatcc ggatgtttgc
     3001 ttccttccac ccttctgcca gcaccgcagg gacctctggg gacagtggac ccccagacag
     3061 gtcacccctg gaacttcaca ttggtttccc cacagacatc cctaaaagtg ccccccactc
     3121 gatgactgcc tcatcttcct cagtttcatc cccatcccca ggtcttccta gacgctcagc
     3181 acccccttct cccctgtgcc gtagtttgtc tcctgggact gggggaggag tccgaggtgg
     3241 ggttggttac ctgtcccgag gggaccctgt ccgggtcctt gctcggagag tacggcctga
     3301 tggctctgtg cagtacctgg ttgagtgggg aggagggggc atcttctgat aatagctcga
     3361 ggcatgcggt accaagcttg tcgagaagta ctagaggatc ataatcagcc ataccacatt
     3421 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3481 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3541 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3601 gtccaaactc atcaatgtat cttatcatgt ctggatctga tcactgatat cgcctaggag
     3661 atccgaacca gataagtgaa atctagttcc aaactatttt gtcattttta attttcgtat
     3721 tagcttacga cgctacaccc agttcccatc tattttgtca ctcttcccta aataatcctt
     3781 aaaaactcca tttccacccc tcccagttcc caactatttt gtccgcccac agcggggcat
     3841 ttttcttcct gttatgtttt taatcaaaca tcctgccaac tccatgtgac aaaccgtcat
     3901 cttcggctac tttttctctg tcacagaatg aaaatttttc tgtcatctct tcgttattaa
     3961 tgtttgtaat tgactgaata tcaacgctta tttgcagcct gaatggcgaa tgggacgcgc
     4021 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4081 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4141 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4201 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4261 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttcttaata gtggactctt
     4321 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4381 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4441 ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga
     4501 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4561 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4621 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4681 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4741 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4801 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4861 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4921 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4981 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5041 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5101 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     5161 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5221 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     5281 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     5341 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5401 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5461 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5521 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5581 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5641 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5701 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5761 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     5821 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5881 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5941 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6001 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6061 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6121 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6181 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6241 ttacggttcc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6301 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6361 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     6421 tctgtgcggt atttcacacc gcatagacca gccgcgtaac ctggcaaaat cggttacggt
     6481 tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg
     6541 aacaaaatag atctaaacta tgacaataaa gtcttaaact agacagaata gttgtaaact
     6601 gaaatcagtc cagttatgct gtgaaaaagc atactggact tttgttatgg ctaaagcaaa
     6661 ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg gccaagggca
     6721 tggtaaagac tatattcgcg gcgttgtgac aatttaccga acaactccgc ggccgggaag
     6781 ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc
     6841 acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc accgtaatct
     6901 gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag gagattgatg
     6961 agcgcggtgg caatgccctg cctccggtgc tcgccggaga ctgcgagatc atagatatag
     7021 atctcactac gcggctgctc aaacttgggc agaacgtaag ccgcgagagc gccaacaacc
     7081 gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa gttcccgagg
     7141 taatcggagt ccggctgatg ttgggagtag gtggctacgt ctccgaactc acgaccgaaa
     7201 agatcaagag cagcccgcat ggatttgact tggtcagggc cgagcctaca tgtgcgaatg
     7261 atgcccatac ttgagccacc taactttgtt ttagggcgac tgccctgctg cgtaacatcg
     7321 ttgctgctgc gtaacatcaa acatcgaccc acggcgtaac gcgcttgctg cttggatgcc
     7381 cgaggcatag actgtacaaa aaaacagtca taacaagcca tgaaaaccgc cactgcgccg
     7441 ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgagcgcat acgctacttg
     7501 cattacagtt tacgaaccga acaggcttat gtcaactggg ttcgtgcctt catccgtttc
     7561 cacggtgtgc gtc
//
