LOCUS       sequence-292442-        8283 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human SUZ12 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 8283)
  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 8283)
  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.SUZ12
FEATURES             Location/Qualifiers
     source          1..8283
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(153..158,159..175,176..181))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        533..624
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     549..566
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             685..702
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             709..1806
                     /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      1663..1672
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1780..1803
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1834..1857
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    4432..4566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4469..4488)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4523..4542
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4595..4760
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4944..5398
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5031..5050)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5240..5261
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5424..5528
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5529..5597,5598..6389)
                     /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(5747..5766)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6560..7148
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7049..7068
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7291..7308
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  7442..7666
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(7733..8266)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg tacaaaaaaa cagtcataac
       61 aagccatgaa aaccgccact gcgccgttac caccgctgcg ttcggtcaag gttctggacc
      121 agttgcgtga gcgcatacgc tacttgcatt acagtttacg aaccgaacag gcttatgtca
      181 actgggttcg tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc ttgggcagca
      241 gcgaagtcga ggcatttctg tcctggctgg cgaacgagcg caaggtttcg gtctccacgc
      301 atcgtcaggc attggcggcc ttgctgttct tctacggcaa ggtgctgtgc acggatctgc
      361 cctggcttca ggagatcgga agacctcggc cgtcgcggcg cttgccggtg gtgctgaccc
      421 cggatgaagt ggttcgcatc ctcggttttc tggaaggcga gcatcgtttg ttcgcccagg
      481 actctagcta tagttctagt ggttggctac gtatactccg gaatattaat agatcatgga
      541 gataattaaa atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt
      601 tttgtaataa aaaaacctat aaatattccg gattattcat accgtcccac catcgggcgc
      661 ggatctcggt ccgaaaccat gggtcatcac catcatcatc acggtaccaa aactgaagaa
      721 ggtaaactgg taatctggat taacggcgat aaaggctata acggtctcgc tgaagtcggt
      781 aagaaattcg agaaagatac cggaattaaa gtcaccgttg agcatccgga taaactggaa
      841 gagaaattcc cacaggttgc ggcaactggc gatggccctg acattatctt ctgggcacac
      901 gaccgctttg gtggctacgc tcaatctggc ctgttggctg aaatcacccc ggacaaagcg
      961 ttccaggaca agctgtatcc gtttacctgg gatgccgtac gttacaacgg caagctgatt
     1021 gcttacccga tcgctgttga agcgttatcg ctgatttata acaaagatct gctgccgaac
     1081 ccgccaaaaa cctgggaaga gatcccggcg ctggataaag aactgaaagc gaaaggtaag
     1141 agcgcgctga tgttcaacct gcaagaaccg tacttcacct ggccgctgat tgctgctgac
     1201 gggggttatg cgttcaagta tgaaaacggc aagtacgaca ttaaagacgt gggcgtggat
     1261 aacgctggcg cgaaagcggg tctgaccttc ctggttgacc tgattaaaaa caaacacatg
     1321 aatgcagaca ccgattactc catcgcagaa gctgccttta ataaaggcga aacagcgatg
     1381 accatcaacg gcccgtgggc atggtccaac atcgacacca gcaaagtgaa ttatggtgta
     1441 acggtactgc cgaccttcaa gggtcaacca tccaaaccgt tcgttggcgt gctgagcgca
     1501 ggtattaacg ccgccagtcc gaacaaagag ctggcaaaag agttcctcga aaactatctg
     1561 ctgactgatg aaggtctgga agcggttaat aaagacaaac cgctgggtgc cgtagcgctg
     1621 aagtcttacg aggaagagtt ggcgaaagat ccacgtattg ccgccaccat ggaaaacgcc
     1681 cagaaaggtg aaatcatgcc gaacatcccg cagatgtccg ctttctggta tgccgtgcgt
     1741 actgcggtga tcaacgccgc cagcggtcgt cagactgtcg atgaagccct gaaagacgcg
     1801 cagactggta ccgattacga tatcccaacg accctggaag ttctgttcca ggggcccctg
     1861 ggatccatgg cgcctcagaa gcacggcggt gggggagggg gcggctcggg gcccagcgcg
     1921 gggtccgggg gaggcggctt cgggggttcg gcggcggtgg cggcggcgac ggcttcgggc
     1981 ggcaaatccg gcggcgggag ctgtggaggg ggtggcagtt actcggcctc ctcctcctcc
     2041 tccgcggcgg cagcggcggg ggctgcggtg ttaccggtga agaagccgaa aatggagcac
     2101 gtccaggctg accacgagct tttcctccag gcctttgaga agccaacaca gatctataga
     2161 tttcttcgaa ctcggaatct catagcacca atatttttgc acagaactct tacttacatg
     2221 tctcatcgaa actccagaac aaacatcaaa aggaaaacat ttaaagttga tgatatgtta
     2281 tcaaaagtag agaaaatgaa aggagagcaa gaatctcata gcttgtcagc tcatttgcag
     2341 cttacgttta ctggtttctt ccacaaaaat gataagccat caccaaactc agaaaatgaa
     2401 caaaattctg ttaccctgga agtcctgctt gtgaaagttt gccacaaaaa aagaaaggat
     2461 gtaagttgtc caataaggca agttcccaca ggtaaaaagc aggtgccttt gaatcctgac
     2521 ctcaatcaaa caaaacccgg aaatttcccg tcccttgcag tttccagtaa tgaatttgaa
     2581 cctagtaaca gccatatggt gaagtcttac tcgttgctat ttagagtgac tcgtccagga
     2641 agaagagagt ttaatggaat gattaatgga gaaaccaatg aaaatattga tgtcaatgaa
     2701 gagcttccag ccagaagaaa acgaaatcgt gaggatgggg aaaagacatt tgttgcacaa
     2761 atgacagtat ttgataaaaa caggcgctta cagcttttag atggggaata tgaagtagcc
     2821 atgcaggaaa tggaagaatg tccaataagc aagaaaagag caacatggga gactattctt
     2881 gatgggaaga ggctgcctcc attcgaaaca ttttctcagg gacctacgtt gcagttcact
     2941 cttcgttgga caggagagac gaatgataaa tctacggctc ctattgccaa acctcttgcc
     3001 actagaaatt cagagagtct ccatcaggaa aacaagcctg gttcagttaa acctactcaa
     3061 actattgctg ttaaagaatc attgactaca gatctacaaa caagaaaaga aaaggatact
     3121 ccaaatgaaa accgacaaaa attaagaata ttttatcagt ttctctataa caacaataca
     3181 aggcaacaaa ctgaagcaag agatgacctg cattgccctt ggtgtactct gaactgccgc
     3241 aaactttata gtttactcaa gcatcttaaa ctctgccata gcagatttat cttcaactat
     3301 gtttatcatc caaaaggtgc taggatagat gtttctatca atgagtgtta tgatggctcc
     3361 tatgcaggaa atcctcagga tattcatcgc caacctggat ttgcttttag tcgcaacgga
     3421 ccagttaaga gaacacctat cacacatatt cttgtgtgca ggccaaaacg aacaaaagca
     3481 agcatgtctg aatttcttga atctgaagat ggggaagtag aacagcaaag aacatatagt
     3541 agtggccaca atcgtctgta tttccatagt gatacctgct tacctctccg tccacaagaa
     3601 atggaagtag atagtgaaga tgaaaaggat cctgaatggc taagagaaaa aaccattaca
     3661 caaattgaag agttttctga tgttaatgaa ggagagaaag aagtgatgaa actctggaat
     3721 ctccatgtca tgaagcatgg gtttattgct gacaatcaaa tgaatcatgc ctgtatgctg
     3781 tttgtagaaa attatggaca gaaaataatt aagaagaatt tatgtcgaaa cttcatgctt
     3841 catctagtca gcatgcatga ctttaatctt attagcataa tgtcaataga taaagctgtt
     3901 accaagctcc gtgaaatgca gcaaaaatta gaaaaggggg aatctgcttc ccctgcaaac
     3961 gaagaaataa ctgaagaaca aaatgggaca gcaaatggat ttagtgaaat taactcaaaa
     4021 gagaaagctt tggaaacaga tagtgtctca ggggtttcaa aacagagcaa aaaacaaaaa
     4081 ctctgaaaag ctctaacccc atgttatgga caaacactga aattacattt tagggaattc
     4141 aaaggcctac gtcgacgagc tcactagtcg cggccgcttt cgaatctaga gcctgcagtc
     4201 tcgaggcatg cggtaccaag cttgtcgaga agtactagag gatcataatc agccatacca
     4261 catttgtgat ctagagcctg cagtctcgag gcatgcggta ccaagcttgt cgagaagtac
     4321 tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     4381 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     4441 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4501 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4561 tggatctgat cactgatatc gcctaggaga tccgaaccag ataagtgaaa tctagttcca
     4621 aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct
     4681 attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc
     4741 aactattttg tccgcccaca gcggggcatt tttcttcctg ttatgttttt aatcaaacat
     4801 cctgccaact ccatgtgaca aaccgtcatc ttcggctact ttttctctgt cacagaatga
     4861 aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat caacgcttat
     4921 ttgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4981 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5041 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5101 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5161 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5221 ggagtccacg ttcttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5281 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5341 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     5401 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5461 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5521 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5581 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5641 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5701 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5761 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5821 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5881 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5941 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6001 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6061 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6121 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6181 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6241 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6301 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6361 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6421 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6481 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6541 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6601 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6661 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6721 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6781 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6841 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6901 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6961 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7021 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7081 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7141 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tcacatgttc
     7201 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7261 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7321 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagaccag
     7381 ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa tggatgccct gcgtaagcgg
     7441 gtgtgggcgg acaataaagt cttaaactga acaaaataga tctaaactat gacaataaag
     7501 tcttaaacta gacagaatag ttgtaaactg aaatcagtcc agttatgctg tgaaaaagca
     7561 tactggactt ttgttatggc taaagcaaac tcttcatttt ctgaagtgca aattgcccgt
     7621 cgtattaaag aggggcgtgg ccaagggcat ggtaaagact atattcgcgg cgttgtgaca
     7681 atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg
     7741 cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac tttgtataga
     7801 gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa gcaccaagcg
     7861 cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc ctccggtgct
     7921 cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca aacttgggca
     7981 gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga
     8041 atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg
     8101 tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt
     8161 ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt
     8221 tagggcgact gccctgctgc gtaacatcgt tgctgctgcg taacatcaaa catcgaccca
     8281 cgg
//
