LOCUS       sequence-449673-        5708 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal hexahistidine-tagged human BMI1 (PCGF4) in 
            insect cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5708)
  AUTHORS   Uckelmann M, Levina V, Taveneau C, Han Ng X, Pandey V, Martinez J, 
            Mendiratta S, Houx J, Boudes M, Venugopal H, Trepout S, Zhang Q, 
            Flanigan S, Li M, Sierecki E, Gambin Y, Das PP, Bell O, de Marco A, 
            Davidovich C
  TITLE     Dynamic PRC1-CBX8 stabilizes a porous structure of chromatin 
            condensates.
  JOURNAL   bioRxiv [Preprint]. 2024 Feb 12:2023.05.08.539931. doi: 
            10.1101/2023.05.08.539931.
  PUBMED    38405976
REFERENCE   2  (bases 1 to 5708)
  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"; journalName: "bioRxiv 
            [Preprint]. 2024 Feb 12:2023.05.08.539931. doi: 
            10.1101/2023.05.08.539931."
COMMENT     Sequence Label: pFBOH-MHL.BMI1
FEATURES             Location/Qualifiers
     source          1..5708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  894..1118
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1185..1718)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1888..1893,1894..1910,1911..1916))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        2268..2359
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2284..2301
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2418..2435
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2448..2468
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     polyA_signal    3581..3715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3618..3637)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3672..3691
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3744..3909
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4093..4547
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4180..4199)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4389..4410
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4573..4677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4678..4746,4747..5538)
                     /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(4896..4915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      781 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagacc agccgcgtaa
      841 cctggcaaaa tcggttacgg ttgagtaata aatggatgcc ctgcgtaagc gggtgtgggc
      901 ggacaataaa gtcttaaact gaacaaaata gatctaaact atgacaataa agtcttaaac
      961 tagacagaat agttgtaaac tgaaatcagt ccagttatgc tgtgaaaaag catactggac
     1021 ttttgttatg gctaaagcaa actcttcatt ttctgaagtg caaattgccc gtcgtattaa
     1081 agaggggcgt ggccaagggc atggtaaaga ctatattcgc ggcgttgtga caatttaccg
     1141 aacaactccg cggccgggaa gccgatctcg gcttgaacga attgttaggt ggcggtactt
     1201 gggtcgatat caaagtgcat cacttcttcc cgtatgccca actttgtata gagagccact
     1261 gcgggatcgt caccgtaatc tgcttgcacg tagatcacat aagcaccaag cgcgttggcc
     1321 tcatgcttga gcagattgat gagcgcggtg gcaatgccct gcctccggtg ctcgccggag
     1381 actgcgagat catagatata gatctcacta cgcggctgct caaacttggg cagaacgtaa
     1441 gccgcgagag cgccaacaac cgcttcttgg tcgaaggcag caagcgcgat gaatgtctta
     1501 ctacggagca agttcccgag gtaatcggag tccggctgat gttgggagta ggtggctacg
     1561 tctccgaact cacgaccgaa aagatcaaga gcagcccgca tggatttgac ttggtcaggg
     1621 ccgagcctac atgtgcgaat gatgcccata cttgagccac ctaactttgt tttagggcga
     1681 ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa
     1741 cgcgcttgct gcttggatgc ccgaggcata gactgtacaa aaaaacagtc ataacaagcc
     1801 atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg tcaaggttct ggaccagttg
     1861 cgtgagcgca tacgctactt gcattacagt ttacgaaccg aacaggctta tgtcaactgg
     1921 gttcgtgcct tcatccgttt ccacggtgtg cgtcacccgg caaccttggg cagcagcgaa
     1981 gtcgaggcat ttctgtcctg gctggcgaac gagcgcaagg tttcggtctc cacgcatcgt
     2041 caggcattgg cggccttgct gttcttctac ggcaaggtgc tgtgcacgga tctgccctgg
     2101 cttcaggaga tcggaagacc tcggccgtcg cggcgcttgc cggtggtgct gaccccggat
     2161 gaagtggttc gcatcctcgg ttttctggaa ggcgagcatc gtttgttcgc ccaggactct
     2221 agctatagtt ctagtggttg gctacgtata ctccggaata ttaatagatc atggagataa
     2281 ttaaaatgat aaccatctcg caaataaata agtattttac tgttttcgta acagttttgt
     2341 aataaaaaaa cctataaata ttccggatta ttcataccgt cccaccatcg ggcgcggatc
     2401 tcggtccgaa aaccatgcat catcatcatc atcacagcag cggcagagaa aacttgtatt
     2461 tccagggcca tcgaacaacg agaatcaaga tcactgagct aaatccccac ctgatgtgtg
     2521 tgctttgtgg agggtacttc attgatgcca caaccataat agaatgtcta cattccttct
     2581 gtaaaacgtg tattgttcgt tacctggaga caagcaagta ttgtcctatt tgtgatgtcc
     2641 aagttcacaa gaccagacca ctactgaata taaggtcaga taaaactctc caagatattg
     2701 tatacaaatt agttccaggg cttttcaaaa atgaaatgaa gagaagaagg gatttttatg
     2761 cagctcatcc ttctgctgat gctgccaatg gctctaatga agatagagga gaggttgcag
     2821 atgaagataa gagaattata actgatgatg agataataag cttatccatt gaattctttg
     2881 accagaacag attggatcgg aaagtaaaca aagacaaaga gaaatctaag gaggaggtga
     2941 atgataaaag atacttacga tgcccagcag caatgactgt gatgcactta agaaagtttc
     3001 tcagaagtaa aatggacata cctaatactt tccagattga tgtcatgtat gaggaggaac
     3061 ctttaaagga ttattataca ctaatggata ttgcctacat ttatacctgg agaaggaatg
     3121 gtccacttcc attgaaatac agagttcgac ctacttgtaa aagaatgaag atcagtcacc
     3181 agagagatgg actgacaaat gctggagaac tggaaagtga ctctgggagt gacaaggcca
     3241 acagcccagc aggaggtatt ccctccacct cttcttgttt gcctagcccc agtactccag
     3301 tgcagtctcc tcatccacag tttcctcaca tttccagtac tatgaatgga accagcaaca
     3361 gccccagcgg taaccaccaa tcttcttttg ccaatagacc tcgaaaatca tcagtaaatg
     3421 ggtcatcagc aacttcttct ggttgatgac gaagcttgtc gagaagtact agaggatcat
     3481 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3541 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3601 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3661 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgatc
     3721 actgatatcg cctaggagat ccgaaccaga taagtgaaat ctagttccaa actattttgt
     3781 catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact
     3841 cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca actattttgt
     3901 ccgcccacag cggggcattt ttcttcctgt tatgttttta atcaaacatc ctgccaactc
     3961 catgtgacaa accgtcatct tcggctactt tttctctgtc acagaatgaa aatttttctg
     4021 tcatctcttc gttattaatg tttgtaattg actgaatatc aacgcttatt tgcagcctga
     4081 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4141 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4201 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4261 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4321 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4381 tcttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     4441 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4501 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
     4561 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4621 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4681 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4741 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4801 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4861 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4921 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4981 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5041 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5101 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5161 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5221 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5281 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5341 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5401 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5461 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5521 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5581 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5641 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5701 ggatcttc
//
