LOCUS       sequence-449677-        6626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal hexahistidine-tagged human CBX8 in insect 
            cells, with a monomeric variant EGFP tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6626)
  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 6626)
  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.mEGFP-CBX8
FEATURES             Location/Qualifiers
     source          1..6626
                     /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"
     CDS             join(2469..2471,2472..2474,2475..3185)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2469..2471,2472..2474,2475..3185)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2514..2535)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2775..2794)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3122..3143
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4499..4633
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4536..4555)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4590..4609
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  4662..4827
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      5011..5465
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5098..5117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5307..5328
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5491..5595
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5596..5664,5665..6456)
                     /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(5814..5833)
                     /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 tccagggcat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     2521 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     2581 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     2641 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     2701 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2761 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2821 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2881 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     2941 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     3001 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3061 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     3121 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     3181 acaagagtgg aagtgagctt tcagcggtgg gggagcgggt gttcgcggcc gaagccctcc
     3241 tgaagcggcg catacggaaa ggacgcatgg aatacctcgt gaaatggaag ggatggtcgc
     3301 agaagtacag cacatgggaa ccggaggaaa acatcctgga tgctcgcttg ctcgcagcct
     3361 ttgaggaaag ggaaagagag atggagctct atggccccaa aaagcgtgga cccaagccca
     3421 aaaccttcct cctcaaagcg caggccaagg caaaggccaa aacttacgag tttcgaagtg
     3481 actcagccag gggcatccgg atcccctacc ctggccgctc gccccaggac ctggcctcca
     3541 cttcccgggc ccgggagggc cttcgaaaca tgggtttgtc cccgccagcg agcagcacca
     3601 gcaccagcag tacctgccgc gcagaggccc ctcgggaccg ggaccgcgac cgggataggg
     3661 accgggagcg ggatcgagaa agggagaggg agcgagagag ggagcgggaa cgtgagaggg
     3721 aacgagagcg gggtaccagc agagtggatg acaagcccag ctcaccgggg gacagctcga
     3781 agaagcgagg ccccaagccc cggaaggagc tcccggaccc ctcacagagg cccttaggcg
     3841 aacccagcgc cggcctcgga gagtacctca agggcaggaa gctggacgac accccttccg
     3901 gggcaggaaa gtttccagcc ggccacagtg tgatccagct ggcccgaaga caggactcgg
     3961 acctggtgca gtgtggtgtg accagcccta gctcagctga ggccacgggc aaactggctg
     4021 tggacacctt cccggccagg gtgataaagc acagggctgc cttcctggag gccaaaggcc
     4081 agggtgccct agatcccaat ggcacccggg tccgacatgg ctcaggcccc cccagctctg
     4141 gggggggcct gtaccgggac atgggggccc aggggggaag gccctccctc atcgccagga
     4201 tccctgtggc cagaatcctg ggggacccgg aggaagagtc ctggagcccc tccctgacta
     4261 acctggagaa ggtggtggtc acggacgtga cctcaaactt tttgaccgtc accattaagg
     4321 aaagtaacac ggaccaaggc ttttttaaag agaaaagatg atgatgacga agcttgtcga
     4381 gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     4441 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     4501 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4561 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4621 tcatgtctgg atctgatcac tgatatcgcc taggagatcc gaaccagata agtgaaatct
     4681 agttccaaac tattttgtca tttttaattt tcgtattagc ttacgacgct acacccagtt
     4741 cccatctatt ttgtcactct tccctaaata atccttaaaa actccatttc cacccctccc
     4801 agttcccaac tattttgtcc gcccacagcg gggcattttt cttcctgtta tgtttttaat
     4861 caaacatcct gccaactcca tgtgacaaac cgtcatcttc ggctactttt tctctgtcac
     4921 agaatgaaaa tttttctgtc atctcttcgt tattaatgtt tgtaattgac tgaatatcaa
     4981 cgcttatttg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     5041 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     5101 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     5161 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     5221 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     5281 tgacgttgga gtccacgttc ttaatagtgg actcttgttc caaactggaa caacactcaa
     5341 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     5401 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     5461 aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5521 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5581 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5641 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5701 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5761 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5821 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5881 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5941 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6001 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6061 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6121 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6181 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6241 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6301 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6361 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6421 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6481 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6541 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6601 cccgtagaaa agatcaaagg atcttc
//
