LOCUS       sequence-449674-        7039 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human CBX8 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 7039)
  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 7039)
  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: pFB1.HMBP.PrS.CBX8
FEATURES             Location/Qualifiers
     source          1..7039
                     /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     732..749
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  883..1107
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(1174..1707)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(1877..1882,1883..1899,1900..1905))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        2255..2346
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     2271..2288
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             2407..2424
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2431..3528
                     /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      3385..3394
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3502..3525
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             3565..3588
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     polyA_signal    4912..5046
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4949..4968)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5003..5022
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5075..5240
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      5424..5878
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5511..5530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5720..5741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5904..6008
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6009..6077,6078..6869)
                     /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(6227..6246)
                     /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 ctcacatgtt ctttcctgcg ttatcccctg
      661 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
      721 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
      781 tccttacgca tctgtgcggt atttcacacc gcatagacca gccgcgtaac ctggcaaaat
      841 cggttacggt tgagtaataa atggatgccc tgcgtaagcg ggtgtgggcg gacaataaag
      901 tcttaaactg aacaaaatag atctaaacta tgacaataaa gtcttaaact agacagaata
      961 gttgtaaact gaaatcagtc cagttatgct gtgaaaaagc atactggact tttgttatgg
     1021 ctaaagcaaa ctcttcattt tctgaagtgc aaattgcccg tcgtattaaa gaggggcgtg
     1081 gccaagggca tggtaaagac tatattcgcg gcgttgtgac aatttaccga acaactccgc
     1141 ggccgggaag ccgatctcgg cttgaacgaa ttgttaggtg gcggtacttg ggtcgatatc
     1201 aaagtgcatc acttcttccc gtatgcccaa ctttgtatag agagccactg cgggatcgtc
     1261 accgtaatct gcttgcacgt agatcacata agcaccaagc gcgttggcct catgcttgag
     1321 gagattgatg agcgcggtgg caatgccctg cctccggtgc tcgccggaga ctgcgagatc
     1381 atagatatag atctcactac gcggctgctc aaacttgggc agaacgtaag ccgcgagagc
     1441 gccaacaacc gcttcttggt cgaaggcagc aagcgcgatg aatgtcttac tacggagcaa
     1501 gttcccgagg taatcggagt ccggctgatg ttgggagtag gtggctacgt ctccgaactc
     1561 acgaccgaaa agatcaagag cagcccgcat ggatttgact tggtcagggc cgagcctaca
     1621 tgtgcgaatg atgcccatac ttgagccacc taactttgtt ttagggcgac tgccctgctg
     1681 cgtaacatcg ttgctgctgc gtaacatcaa acatcgaccc acggcgtaac gcgcttgctg
     1741 cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca tgaaaaccgc
     1801 cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgagcgcat
     1861 acgctacttg cattacagtt tacgaaccga acaggcttat gtcaactggg ttcgtgcctt
     1921 catccgtttc cacggtgtgc gtcacccggc aaccttgggc agcagcgaag tcgaggcatt
     1981 tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc acgcatcgtc aggcattggc
     2041 ggccttgctg ttcttctacg gcaaggtgct gtgcacggat ctgccctggc ttcaggagat
     2101 cggaagacct cggccgtcgc ggcgcttgcc ggtggtgctg accccggatg aagtggttcg
     2161 catcctcggt tttctggaag gcgagcatcg tttgttcgcc caggactcta gctatagttc
     2221 tagtggttgg ctacatactc cggaatatta atagatcatg gagataatta aaatgataac
     2281 catctcgcaa ataaataagt attttactgt tttcgtaaca gttttgtaat aaaaaaacct
     2341 ataaatattc cggattattc ataccgtccc accatcgggc gcggatctcg gtccgaaacc
     2401 atgggtcatc accatcatca tcacggtacc aaaactgaag aaggtaaact ggtaatctgg
     2461 attaacggcg ataaaggcta taacggtctc gctgaagtcg gtaagaaatt cgagaaagat
     2521 accggaatta aagtcaccgt tgagcatccg gataaactgg aagagaaatt cccacaggtt
     2581 gcggcaactg gcgatggccc tgacattatc ttctgggcac acgaccgctt tggtggctac
     2641 gctcaatctg gcctgttggc tgaaatcacc ccggacaaag cgttccagga caagctgtat
     2701 ccgtttacct gggatgccgt acgttacaac ggcaagctga ttgcttaccc gatcgctgtt
     2761 gaagcgttat cgctgattta taacaaagat ctgctgccga acccgccaaa aacctgggaa
     2821 gagatcccgg cgctggataa agaactgaaa gcgaaaggta agagcgcgct gatgttcaac
     2881 ctgcaagaac cgtacttcac ctggccgctg attgctgctg acgggggtta tgcgttcaag
     2941 tatgaaaacg gcaagtacga cattaaagac gtgggcgtgg ataacgctgg cgcgaaagcg
     3001 ggtctgacct tcctggttga cctgattaaa aacaaacaca tgaatgcaga caccgattac
     3061 tccatcgcag aagctgcctt taataaaggc gaaacagcga tgaccatcaa cggcccgtgg
     3121 gcatggtcca acatcgacac cagcaaagtg aattatggtg taacggtact gccgaccttc
     3181 aagggtcaac catccaaacc gttcgttggc gtgctgagcg caggtattaa cgccgccagt
     3241 ccgaacaaag agctggcaaa agagttcctc gaaaactatc tgctgactga tgaaggtctg
     3301 gaagcggtta ataaagacaa accgctgggt gccgtagcgc tgaagtctta cgaggaagag
     3361 ttggcgaaag atccacgtat tgccgccacc atggaaaacg cccagaaagg tgaaatcatg
     3421 ccgaacatcc cgcagatgtc cgctttctgg tatgccgtgc gtactgcggt gatcaacgcc
     3481 gccagcggtc gtcagactgt cgatgaagcc ctgaaagacg cgcagactgc ggccgcaggc
     3541 accgattacg atatcccaac gaccctggaa gttctgttcc aggggcccgg ggagctttca
     3601 gcggtggggg agcgggtgtt cgcggccgaa gccctcctga agcggcgcat acggaaagga
     3661 cgcatggaat acctcgtgaa atggaaggga tggtcgcaga agtacagcac atgggaaccg
     3721 gaggaaaaca tcctggatgc tcgcttgctc gcagcctttg aggaaaggga aagagagatg
     3781 gagctctatg gccccaaaaa gcgtggaccc aagcccaaaa ccttcctcct caaagcgcag
     3841 gccaaggcaa aggccaaaac ttacgagttt cgaagtgact cagccagggg catccggatc
     3901 ccctaccctg gccgctcgcc ccaggacctg gcctccactt cccgggcccg ggagggcctt
     3961 cgaaacatgg gtttgtcccc gccagcgagc agcaccagca ccagcagtac ctgccgcgca
     4021 gaggcccctc gggaccggga ccgcgaccgg gatagggacc gggagcggga tcgagaaagg
     4081 gagagggagc gagagaggga gcgggaacgt gagagggaac gagagcgggg taccagcaga
     4141 gtggatgaca agcccagctc accgggggac agctcgaaga agcgaggccc caagccccgg
     4201 aaggagctcc cggacccctc acagaggccc ttaggcgaac ccagcgccgg cctcggagag
     4261 tacctcaagg gcaggaagct ggacgacacc ccttccgggg caggaaagtt tccagccggc
     4321 cacagtgtga tccagctggc ccgaagacag gactcggacc tggtgcagtg tggtgtgacc
     4381 agccctagct cagctgaggc cacgggcaaa ctggctgtgg acaccttccc ggccagggtg
     4441 ataaagcaca gggctgcctt cctggaggcc aaaggccagg gtgccctaga tcccaatggc
     4501 acccgggtcc gacatggctc aggccccccc agctctgggg ggggcctgta ccgggacatg
     4561 ggggcccagg ggggaaggcc ctccctcatc gccaggatcc ctgtggccag aatcctgggg
     4621 gacccggagg aagagtcctg gagcccctcc ctgactaacc tggagaaggt ggtggtcacg
     4681 gacgtgacct caaacttttt gaccgtcacc attaaggaaa gtaacacgga ccaaggcttt
     4741 tttaaagaga aaagatgata atagctcgag gcatgcggta ccaagcttgt cgagaagtac
     4801 tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     4861 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     4921 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4981 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5041 tggatctgat cactgatatc gcctaggaga tccgaaccag ataagtgaaa tctagttcca
     5101 aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct
     5161 attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc
     5221 aactattttg tccgcccaca gcggggcatt tttcttcctg ttatgttttt aatcaaacat
     5281 cctgccaact ccatgtgaca aaccgtcatc ttcggctact ttttctctgt cacagaatga
     5341 aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat caacgcttat
     5401 ttgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5461 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5521 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5581 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5641 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5701 ggagtccacg ttcttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5761 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5821 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     5881 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5941 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6001 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6061 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6121 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6181 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6241 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6301 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6361 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6421 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6481 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6541 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6601 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6661 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6721 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6781 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6841 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6901 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6961 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7021 aaaagatcaa aggatcttc
//
