LOCUS       sequence-292456-        7389 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human AEBP2 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 7389)
  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 7389)
  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.AEBP2
FEATURES             Location/Qualifiers
     source          1..7389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        142..233
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     158..175
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             294..311
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             318..1415
                     /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      1272..1281
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1389..1412
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1443..1466
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    3147..3281
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3184..3203)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3238..3257
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3310..3475
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3659..4113
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3746..3765)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3955..3976
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4139..4243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4244..4312,4313..5104)
                     /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(4462..4481)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5275..5863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5764..5783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6006..6023
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6157..6381
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6448..6981)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7151..7156,7157..7173,7174..7179))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 gtcgcggcgc ttgccggtgg tgctgacccc ggatgaagtg gttcgcatcc tcggttttct
       61 ggaaggcgag catcgtttgt tcgcccagga ctctagctat agttctagtg gttggctacg
      121 tatactccgg aatattaata gatcatggag ataattaaaa tgataaccat ctcgcaaata
      181 aataagtatt ttactgtttt cgtaacagtt ttgtaataaa aaaacctata aatattccgg
      241 attattcata ccgtcccacc atcgggcgcg gatctcggtc cgaaaccatg ggtcatcacc
      301 atcatcatca cggtaccaaa actgaagaag gtaaactggt aatctggatt aacggcgata
      361 aaggctataa cggtctcgct gaagtcggta agaaattcga gaaagatacc ggaattaaag
      421 tcaccgttga gcatccggat aaactggaag agaaattccc acaggttgcg gcaactggcg
      481 atggccctga cattatcttc tgggcacacg accgctttgg tggctacgct caatctggcc
      541 tgttggctga aatcaccccg gacaaagcgt tccaggacaa gctgtatccg tttacctggg
      601 atgccgtacg ttacaacggc aagctgattg cttacccgat cgctgttgaa gcgttatcgc
      661 tgatttataa caaagatctg ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc
      721 tggataaaga actgaaagcg aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt
      781 acttcacctg gccgctgatt gctgctgacg ggggttatgc gttcaagtat gaaaacggca
      841 agtacgacat taaagacgtg ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc
      901 tggttgacct gattaaaaac aaacacatga atgcagacac cgattactcc atcgcagaag
      961 ctgcctttaa taaaggcgaa acagcgatga ccatcaacgg cccgtgggca tggtccaaca
     1021 tcgacaccag caaagtgaat tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat
     1081 ccaaaccgtt cgttggcgtg ctgagcgcag gtattaacgc cgccagtccg aacaaagagc
     1141 tggcaaaaga gttcctcgaa aactatctgc tgactgatga aggtctggaa gcggttaata
     1201 aagacaaacc gctgggtgcc gtagcgctga agtcttacga ggaagagttg gcgaaagatc
     1261 cacgtattgc cgccaccatg gaaaacgccc agaaaggtga aatcatgccg aacatcccgc
     1321 agatgtccgc tttctggtat gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc
     1381 agactgtcga tgaagccctg aaagacgcgc agactggtac cgattacgat atcccaacga
     1441 ccctggaagt tctgttccag gggcccctgg gatccgccgc cgctatcacc gacatggccg
     1501 acctggagga gctctcccgc ctgagccctc tgccccccgg cagcccgggt tcggcggcgc
     1561 ggggccgggc tgagcccccc gaggaggagg aggaagagga ggaggaggaa gaggaggcgg
     1621 aggccgaggc ggtggcggcg ctgctgctga acggcggcag cggtgggggc ggcggaggcg
     1681 gcggcggagg agtggggggc ggcgaggcag agacgatgtc ggagccgagc cccgagagcg
     1741 ccagccaggc cggggaggac gaagacgagg aggaggacga cgaggaggag gaagatgaga
     1801 gcagcagcag cggcgggggt gaggaggaga gtagcgccga gagcctggtg ggcagcagcg
     1861 gcgggagcag cagcgacgag acccgctcgt tgagccccgg cgccgccagc agcagcagcg
     1921 gggatgggga cggcaaggag ggcctggagg agcccaaggg accgcggggc agccagggcg
     1981 gcggcggggg cggcagcagt agcagcagcg tagtctccag cggcggcgac gagggctacg
     2041 ggactggggg aggcggaagc agcgcgacct ccgggggccg gcggggcagc ttggagatgt
     2101 cgtcggatgg ggaacccctg agccgcatgg actcggagga cagcataagc agtactataa
     2161 tggatgtaga cagcacaatt tccagtgggc gttcaactcc agcaatgatg aatggacaag
     2221 gaagcactac ttcttcaagc aaaaatattg cctataattg ttgttgggac cagtgccagg
     2281 cttgcttcaa ctctagccca gatctggcag atcacatccg ttccatacat gtagatggtc
     2341 agcgaggagg ggtatttgtt tgcttatgga aaggttgtaa agtatataac actccatcta
     2401 ccagtcaaag ttggttacaa aggcatatgc tgacacacag tggagacaaa cctttcaagt
     2461 gtgttgttgg tggctgcaat gccagctttg cttctcaggg agggctagct cgtcatgtac
     2521 ccacacactt cagtcagcag aactcctcaa aagtttctag ccagccaaag gccaaagaag
     2581 aatctccttc taaagctgga atgaacaaaa ggaggaaatt aaagaacaaa agacgacgct
     2641 cattaccacg gccacatgat ttcttcgatg cacaaacact ggatgcgata agacatcgag
     2701 ccatatgctt taacctctca gctcatatag aaagtttagg gaagggacac agtgttgttt
     2761 ttcatagtac tgtaatagct aagagaaaag aagattctgg gaagatcaaa cttttgcttc
     2821 attggatgcc tgaagacatt ctgcctgatg tgtgggtgaa tgaaagtgaa cgacatcagt
     2881 taaaaactaa agtagttcat ttatcaaagc tacccaaaga tactgccttg cttttggacc
     2941 caaacatata cagaacaatg ccgcagaaga ggttgaagag gtgatctaga gcctgcagtc
     3001 tcgaggcatg cggtaccaag cttgtcgaga agtactagag gatcataatc agccatacca
     3061 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     3121 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     3181 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3241 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg atatcgccta
     3301 ggagatccga accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc
     3361 gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc cctaaataat
     3421 ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc ccacagcggg
     3481 gcatttttct tcctgttatg tttttaatca aacatcctgc caactccatg tgacaaaccg
     3541 tcatcttcgg ctactttttc tctgtcacag aatgaaaatt tttctgtcat ctcttcgtta
     3601 ttaatgtttg taattgactg aatatcaacg cttatttgca gcctgaatgg cgaatgggac
     3661 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3721 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3781 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3841 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3901 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt aatagtggac
     3961 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4021 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4081 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     4141 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4201 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4261 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4321 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4381 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4441 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4501 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4561 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4621 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4681 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4741 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4801 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4861 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4921 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4981 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5041 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5101 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5161 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5221 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5281 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5341 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5401 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5461 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5521 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5581 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5641 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5701 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5761 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5821 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5881 ctttttacgg ttcctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     5941 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     6001 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     6061 cgcatctgtg cggtatttca caccgcatag accagccgcg taacctggca aaatcggtta
     6121 cggttgagta ataaatggat gccctgcgta agcgggtgtg ggcggacaat aaagtcttaa
     6181 actgaacaaa atagatctaa actatgacaa taaagtctta aactagacag aatagttgta
     6241 aactgaaatc agtccagtta tgctgtgaaa aagcatactg gacttttgtt atggctaaag
     6301 caaactcttc attttctgaa gtgcaaattg cccgtcgtat taaagagggg cgtggccaag
     6361 ggcatggtaa agactatatt cgcggcgttg tgacaattta ccgaacaact ccgcggccgg
     6421 gaagccgatc tcggcttgaa cgaattgtta ggtggcggta cttgggtcga tatcaaagtg
     6481 catcacttct tcccgtatgc ccaactttgt atagagagcc actgcgggat cgtcaccgta
     6541 atctgcttgc acgtagatca cataagcacc aagcgcgttg gcctcatgct tgaggagatt
     6601 gatgagcgcg gtggcaatgc cctgcctccg gtgctcgccg gagactgcga gatcatagat
     6661 atagatctca ctacgcggct gctcaaactt gggcagaacg taagccgcga gagcgccaac
     6721 aaccgcttct tggtcgaagg cagcaagcgc gatgaatgtc ttactacgga gcaagttccc
     6781 gaggtaatcg gagtccggct gatgttggga gtaggtggct acgtctccga actcacgacc
     6841 gaaaagatca agagcagccc gcatggattt gacttggtca gggccgagcc tacatgtgcg
     6901 aatgatgccc atacttgagc cacctaactt tgttttaggg cgactgccct gctgcgtaac
     6961 atcgttgctg ctgcgtaaca tcaaacatcg acccacggcg taacgcgctt gctgcttgga
     7021 tgcccgaggc atagactgta caaaaaaaca gtcataacaa gccatgaaaa ccgccactgc
     7081 gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc gcatacgcta
     7141 cttgcattac agtttacgaa ccgaacaggc ttatgtcaac tgggttcgtg ccttcatccg
     7201 tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg catttctgtc
     7261 ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat tggcggcctt
     7321 gctgttcttc tacggcaagg tgctgtgcac ggatctgccc tggcttcagg agatcggaag
     7381 acctcggcc
//
