LOCUS       sequence-293826-        6005 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human RBBP4 in insect cells, under a TEV-cleavable 
            N-terminal polyhistidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6005)
  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 6005)
  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: pFBOH-MHL.RBBP4
FEATURES             Location/Qualifiers
     source          1..6005
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        177..268
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     193..210
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             327..344
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             357..377
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    1787..1921
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1824..1843)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1878..1897
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  1950..2115
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      2299..2753
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2386..2405)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2595..2616
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2779..2883
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2884..2952,2953..3744)
                     /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(3102..3121)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3915..4503
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4404..4423
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4657..4674
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  4808..5032
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(5099..5632)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(5802..5807,5808..5824,5825..5830))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 ctgccctggc ttcaggagat cggaagacct cggccgtcgc ggcgcttgcc ggtggtgctg
       61 accccggatg aagtggttcg catcctcggt tttctggaag gcgagcatcg tttgttcgcc
      121 caggactcta gctatagttc tagtggttgg ctacgtatac tccggaatat taatagatca
      181 tggagataat taaaatgata accatctcgc aaataaataa gtattttact gttttcgtaa
      241 cagttttgta ataaaaaaac ctataaatat tccggattat tcataccgtc ccaccatcgg
      301 gcgcggatct cggtccgaaa accatgcatc atcatcatca tcacagcagc ggcagagaaa
      361 acttgtattt ccagggcgcc gacaaggaag cagccttcga cgacgcagtg gaagaacgag
      421 tgatcaacga ggaatacaaa atatggaaaa agaacacccc ttttctttat gatttggtga
      481 tgacccatgc tctggagtgg cccagcctaa ctgcccagtg gcttccagat gtaaccagac
      541 cagaagggaa agatttcagc attcatcgac ttgtcctggg gacacacaca tcggatgaac
      601 aaaaccatct tgttatagcc agtgtgcagc tccctaatga tgatgctcag tttgatgcgt
      661 cacactacga cagtgagaaa ggagaatttg gaggttttgg ttcagttagt ggaaaaattg
      721 aaatagaaat caagatcaac catgaaggag aagtaaacag ggcccgttat atgccccaga
      781 acccttgtat catcgcaaca aagactcctt ccagtgatgt tcttgtcttt gactatacaa
      841 aacatccttc taaaccagat ccttctggag agtgcaaccc agacttgcgt ctccgtggac
      901 atcagaagga aggctatggg ctttcttgga acccaaatct cagtgggcac ttacttagtg
      961 cttcagatga ccataccatc tgcctgtggg acatcagtgc cgttccaaag gagggaaaag
     1021 tggtagatgc gaagaccatc tttacagggc atacggcagt agtagaagat gtttcctggc
     1081 atctactcca tgagtctctg tttgggtcag ttgctgatga tcagaaactt atgatttggg
     1141 atactcgttc aaacaatact tccaaaccaa gccactcagt tgatgctcac actgctgaag
     1201 tgaactgcct ttctttcaat ccttatagtg agttcattct tgccacagga tcagctgaca
     1261 agactgttgc cttgtgggat ctgagaaatc tgaaacttaa gttgcattcc tttgagtcac
     1321 ataaggatga aatattccag gttcagtggt cacctcacaa tgagactatt ttagcttcca
     1381 gtggtactga tcgcagactg aatgtctggg atttaagtaa aattggagag gaacaatccc
     1441 cagaagatgc agaagacggg ccaccagagt tgttgtttat tcatggtggt catactgcca
     1501 agatatctga tttctcctgg aatcccaatg aaccttgggt gatttgttct gtatcagaag
     1561 acaatatcat gcaagtgtgg caaatggcag agaacattta taatgatgaa gaccctgaag
     1621 gaagcgtgga tccagaagga caagggtcct gatgacgaag cttgtcgaga agtactagag
     1681 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     1741 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     1801 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1861 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat
     1921 ctgatcactg atatcgccta ggagatccga accagataag tgaaatctag ttccaaacta
     1981 ttttgtcatt tttaattttc gtattagctt acgacgctac acccagttcc catctatttt
     2041 gtcactcttc cctaaataat ccttaaaaac tccatttcca cccctcccag ttcccaacta
     2101 ttttgtccgc ccacagcggg gcatttttct tcctgttatg tttttaatca aacatcctgc
     2161 caactccatg tgacaaaccg tcatcttcgg ctactttttc tctgtcacag aatgaaaatt
     2221 tttctgtcat ctcttcgtta ttaatgtttg taattgactg aatatcaacg cttatttgca
     2281 gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2341 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2401 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2461 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2521 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2581 ccacgttctt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     2641 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2701 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     2761 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     2821 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     2881 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     2941 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3001 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3061 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3121 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3181 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3241 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3301 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3361 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3421 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3481 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3541 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3601 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3661 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3721 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     3781 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     3841 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     3901 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     3961 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     4021 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     4081 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     4141 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     4201 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     4261 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     4321 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     4381 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     4441 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     4501 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     4561 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     4621 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     4681 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4741 gaccagccgc gtaacctggc aaaatcggtt acggttgagt aataaatgga tgccctgcgt
     4801 aagcgggtgt gggcggacaa taaagtctta aactgaacaa aatagatcta aactatgaca
     4861 ataaagtctt aaactagaca gaatagttgt aaactgaaat cagtccagtt atgctgtgaa
     4921 aaagcatact ggacttttgt tatggctaaa gcaaactctt cattttctga agtgcaaatt
     4981 gcccgtcgta ttaaagaggg gcgtggccaa gggcatggta aagactatat tcgcggcgtt
     5041 gtgacaattt accgaacaac tccgcggccg ggaagccgat ctcggcttga acgaattgtt
     5101 aggtggcggt acttgggtcg atatcaaagt gcatcacttc ttcccgtatg cccaactttg
     5161 tatagagagc cactgcggga tcgtcaccgt aatctgcttg cacgtagatc acataagcac
     5221 caagcgcgtt ggcctcatgc ttgagcagat tgatgagcgc ggtggcaatg ccctgcctcc
     5281 ggtgctcgcc ggagactgcg agatcataga tatagatctc actacgcggc tgctcaaact
     5341 tgggcagaac gtaagccgcg agagcgccaa caaccgcttc ttggtcgaag gcagcaagcg
     5401 cgatgaatgt cttactacgg agcaagttcc cgaggtaatc ggagtccggc tgatgttggg
     5461 agtaggtggc tacgtctccg aactcacgac cgaaaagatc aagagcagcc cgcatggatt
     5521 tgacttggtc agggccgagc ctacatgtgc gaatgatgcc catacttgag ccacctaact
     5581 ttgttttagg gcgactgccc tgctgcgtaa catcgttgct gctccataac atcaaacatc
     5641 gacccacggc gtaacgcgct tgctgcttgg atgcccgagg catagactgt acaaaaaaac
     5701 agtcataaca agccatgaaa accgccactg cgccgttacc accgctgcgt tcggtcaagg
     5761 ttctggacca gttgcgtgag cgcatacgct acttgcatta cagtttacga accgaacagg
     5821 cttatgtcaa ctgggttcgt gccttcatcc gtttccacgg tgtgcgtcac ccggcaacct
     5881 tgggcagcag cgaagtcgag gcatttctgt cctggctggc gaacgagcgc aaggtttcgg
     5941 tctccacgca tcgtcaggca ttggcggcct tgctgttctt ctacggcaag gtgctgtgca
     6001 cggat
//
