LOCUS       sequence-292458-        7651 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human MTF2 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 7651)
  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 7651)
  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.MTF2
FEATURES             Location/Qualifiers
     source          1..7651
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        186..277
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     202..219
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             338..355
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             362..1459
                     /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      1316..1325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1433..1456
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1496..1519
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    3455..3589
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3492..3511)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3546..3565
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3618..3783
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3967..4421
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4054..4073)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4263..4284
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4447..4551
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4552..4620,4621..5412)
                     /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(4770..4789)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5583..6171
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6072..6091
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6314..6331
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6465..6689
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6756..7289)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7459..7464,7465..7481,7482..7487))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 tgtgcacgga tctgccctgg cttcaggaga tcggaagacc tcggccgtcg cggcgcttgc
       61 cggtggtgct gaccccggat gaagtggttc gcatcctcgg ttttctggaa ggcgagcatc
      121 gtttgttcgc ccaggactct agctatagtt ctagtggttg gctacatact ccggaatatt
      181 aatagatcat ggagataatt aaaatgataa ccatctcgca aataaataag tattttactg
      241 ttttcgtaac agttttgtaa taaaaaaacc tataaatatt ccggattatt cataccgtcc
      301 caccatcggg cgcggatctc ggtccgaaac catgggtcat caccatcatc atcacggtac
      361 caaaactgaa gaaggtaaac tggtaatctg gattaacggc gataaaggct ataacggtct
      421 cgctgaagtc ggtaagaaat tcgagaaaga taccggaatt aaagtcaccg ttgagcatcc
      481 ggataaactg gaagagaaat tcccacaggt tgcggcaact ggcgatggcc ctgacattat
      541 cttctgggca cacgaccgct ttggtggcta cgctcaatct ggcctgttgg ctgaaatcac
      601 cccggacaaa gcgttccagg acaagctgta tccgtttacc tgggatgccg tacgttacaa
      661 cggcaagctg attgcttacc cgatcgctgt tgaagcgtta tcgctgattt ataacaaaga
      721 tctgctgccg aacccgccaa aaacctggga agagatcccg gcgctggata aagaactgaa
      781 agcgaaaggt aagagcgcgc tgatgttcaa cctgcaagaa ccgtacttca cctggccgct
      841 gattgctgct gacgggggtt atgcgttcaa gtatgaaaac ggcaagtacg acattaaaga
      901 cgtgggcgtg gataacgctg gcgcgaaagc gggtctgacc ttcctggttg acctgattaa
      961 aaacaaacac atgaatgcag acaccgatta ctccatcgca gaagctgcct ttaataaagg
     1021 cgaaacagcg atgaccatca acggcccgtg ggcatggtcc aacatcgaca ccagcaaagt
     1081 gaattatggt gtaacggtac tgccgacctt caagggtcaa ccatccaaac cgttcgttgg
     1141 cgtgctgagc gcaggtatta acgccgccag tccgaacaaa gagctggcaa aagagttcct
     1201 cgaaaactat ctgctgactg atgaaggtct ggaagcggtt aataaagaca aaccgctggg
     1261 tgccgtagcg ctgaagtctt acgaggaaga gttggcgaaa gatccacgta ttgccgccac
     1321 catggaaaac gcccagaaag gtgaaatcat gccgaacatc ccgcagatgt ccgctttctg
     1381 gtatgccgtg cgtactgcgg tgatcaacgc cgccagcggt cgtcagactg tcgatgaagc
     1441 cctgaaagac gcgcagactg cggccgcagg caccgattac gatatcccaa cgaccctgga
     1501 agttctgttc caggggcccg ggagagactc tacaggggca ggtaattcac tggtccacaa
     1561 gcggtcgcct ttacgtcgaa accaaaagac cccaacatcc ttgaccaagc tgtctttaca
     1621 ggatggacat aaagccaaaa agccagcatg taaatttgaa gagggtcagg atgtcctagc
     1681 tagatggtca gatggcttgt tttatcttgg cactatcaaa aagataaaca tattgaaaca
     1741 gagctgcttc atcatatttg aagacagttc taaatcctgg gttctctgga aggacattca
     1801 aacaggagcc actggaagtg gggaaatggt ctgtacaata tgtcaagaag agtattcaga
     1861 agctcccaat gaaatggtta tatgtgacaa gtgtggccaa ggatatcatc agttgtgtca
     1921 cacacctcat attgattcca gtgtgattga ttcagatgaa aaatggctct gtcggcagtg
     1981 tgtttttgca acaacaacaa agaggggtgg tgcacttaag aaaggaccaa atgccaaagc
     2041 attgcaagtc atgaagcaga cattacccta tagtgtggca gaccttgaat gggatgcagg
     2101 tcataaaacc aatgtccagc agtgttactg ctattgtgga ggccctggag actggtattt
     2161 gaagatgcta cagtgctgca aatgtaagca gtggtttcat gaggcttgtg tgcaatgcct
     2221 tcaaaagcca atgctatttg gagacagatt ttatacgttt atatgctctg tctgcagttc
     2281 tggaccagaa tacctcaaac gtctaccatt acagtgggta gatatagcac acctatgcct
     2341 ttacaaccta agtgttattc ataagaagaa atactttgat tctgaacttg agcttatgac
     2401 atacattaat gaaaactggg atagattgca ccctggagag ctggcagaca caccaaaatc
     2461 tgaaagatat gagcatgttc tggaggcatt aaatgattac aagaccatgt ttatgtctgg
     2521 gaaagaaata aagaagaaga agcatttgtt tgggttgcga attcgtgttc ctcctgtgcc
     2581 accaaatgtg gctttcaaag cagagaaaga acctgaagga acatctcatg aatttaaaat
     2641 taaaggcaga aaggcatcca aacctatatc tgattcaagg gaagtaagca atggcataga
     2701 aaaaaaagga aagaaaaaat ctgtaggtcg tccacctggc ccatatacaa gaaaaatgat
     2761 tcaaaaaact gctgagccac ttttggataa ggaatcaatt tcagagaatc ctactttgga
     2821 tttaccttgt tctataggga gaactgaggg aactgcacat tcatccaata cctcagatgt
     2881 ggatttcacg ggtgcttcca gtgcaaaaga aactacctcg tctagcattt ccaggcatta
     2941 tggattatct gactccagaa aaagaacgcg tacaggaaga tcttggcctg ctgcaatacc
     3001 acatttgcgg agaagaagag gtcgtcttcc aagaagagca ctccagactc agaactcaga
     3061 aattgtaaaa gatgatgaag gcaaagaaga ttatcagttt gatgaactca acacagagat
     3121 tctgaataac ttagcagatc aggagttaca actcaatcat ctaaagaact ccattaccag
     3181 ttattttggt gctgcaggta gaatagcatg tggcgaaaaa taccgagttt tggcacgtcg
     3241 ggtgacactt gatggaaagg tgcagtatct tgtggaatgg gaaggagcaa ctgcatcctg
     3301 ataatagctc gaggcatgcg gtaccaagct tgtcgagaag tactagagga tcataatcag
     3361 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3421 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3481 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3541 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct gatcactgat
     3601 atcgcctagg agatccgaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt
     3661 taattttcgt attagcttac gacgctacac ccagttccca tctattttgt cactcttccc
     3721 taaataatcc ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc
     3781 acagcggggc atttttcttc ctgttatgtt tttaatcaaa catcctgcca actccatgtg
     3841 acaaaccgtc atcttcggct actttttctc tgtcacagaa tgaaaatttt tctgtcatct
     3901 cttcgttatt aatgtttgta attgactgaa tatcaacgct tatttgcagc ctgaatggcg
     3961 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4021 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4081 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4141 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4201 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttaa
     4261 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4321 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4381 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     4441 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4501 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     4561 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     4621 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     4681 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     4741 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     4801 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     4861 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     4921 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     4981 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5041 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5101 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5161 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5221 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5281 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5341 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5401 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5461 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     5521 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     5581 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     5641 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     5701 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     5761 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5821 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5881 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5941 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6001 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6061 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6121 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6181 aacgcggcct ttttacggtt cctggccttt tgctcacatg ttctttcctg cgttatcccc
     6241 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     6301 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     6361 tctccttacg catctgtgcg gtatttcaca ccgcatagac cagccgcgta acctggcaaa
     6421 atcggttacg gttgagtaat aaatggatgc cctgcgtaag cgggtgtggg cggacaataa
     6481 agtcttaaac tgaacaaaat agatctaaac tatgacaata aagtcttaaa ctagacagaa
     6541 tagttgtaaa ctgaaatcag tccagttatg ctgtgaaaaa gcatactgga cttttgttat
     6601 ggctaaagca aactcttcat tttctgaagt gcaaattgcc cgtcgtatta aagaggggcg
     6661 tggccaaggg catggtaaag actatattcg cggcgttgtg acaatttacc gaacaactcc
     6721 gcggccggga agccgatctc ggcttgaacg aattgttagg tggcggtact tgggtcgata
     6781 tcaaagtgca tcacttcttc ccgtatgccc aactttgtat agagagccac tgcgggatcg
     6841 tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg
     6901 aggagattga tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga gactgcgaga
     6961 tcatagatat agatctcact acgcggctgc tcaaacttgg gcagaacgta agccgcgaga
     7021 gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt actacggagc
     7081 aagttcccga ggtaatcgga gtccggctga tgttgggagt aggtggctac gtctccgaac
     7141 tcacgaccga aaagatcaag agcagcccgc atggatttga cttggtcagg gccgagccta
     7201 catgtgcgaa tgatgcccat acttgagcca cctaactttg ttttagggcg actgccctgc
     7261 tgcgtaacat cgttgctgct gcgtaacatc aaacatcgac ccacggcgta acgcgcttgc
     7321 tgcttggatg cccgaggcat agactgtaca aaaaaacagt cataacaagc catgaaaacc
     7381 gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc
     7441 atacgctact tgcattacag tttacgaacc gaacaggctt atgtcaactg ggttcgtgcc
     7501 ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga agtcgaggca
     7561 tttctgtcct ggctggcgaa cgagcgcaag gtttcggtct ccacgcatcg tcaggcattg
     7621 gcggccttgc tgttcttcta cggcaaggtg c
//
