LOCUS       sequence-449675-        5738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminal hexahistidine-tagged human RING1B (RNF2) in 
            insect cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5738)
  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 5738)
  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.RING1B
FEATURES             Location/Qualifiers
     source          1..5738
                     /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    3611..3745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3648..3667)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3702..3721
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3774..3939
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      4123..4577
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4210..4229)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4419..4440
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4603..4707
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4708..4776,4777..5568)
                     /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(4926..4945)
                     /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 tccagggctc tcaggctgtg cagacaaacg gaactcaacc attaagcaaa acatgggaac
     2521 tcagtttata tgagttacaa cgaacacctc aggaggcaat aacagatggc ttagaaattg
     2581 tggtttcacc tcgaagtcta cacagtgaat taatgtgccc aatttgtttg gatatgttga
     2641 agaacaccat gactacaaag gagtgtttac atcgtttttg tgcagactgc atcatcacag
     2701 cccttagaag tggcaacaaa gaatgtccta cctgtcggaa aaaactagtt tccaaaagat
     2761 cactaaggcc agacccaaac tttgatgcac tcatcagcaa aatttatcca agtcgtgatg
     2821 agtatgaagc tcatcaagag agagtattag ccaggatcaa caagcacaat aatcagcaag
     2881 cactcagtca cagcattgag gaaggactga agatacaggc catgaacaga ctgcagcgag
     2941 gcaagaaaca acagattgaa aatggtagtg gagcagaaga taatggtgac agttcacact
     3001 gcagtaatgc atccacacat agcaatcagg aagcaggccc tagtaacaaa cggaccaaaa
     3061 catctgatga ttctgggcta gagcttgata ataacaatgc agcaatggca attgatccag
     3121 taatggatgg tgctagtgaa attgaattag tattcaggcc tcatcccaca cttatggaaa
     3181 aagatgacag tgcacagacg agatacataa agacttctgg taacgccact gttgatcact
     3241 tatccaagta tctggctgtg aggttagctt tagaagaact tcgaagcaaa ggtgaatcaa
     3301 accagatgaa ccttgataca gccagtgaga agcagtatac catttatata gcaacagcca
     3361 gtggccagtt cactgtatta aatggctctt tttctttgga attggtcagt gagaaatact
     3421 ggaaagtgaa caaacccatg gaactttatt acgcacctac aaaggagcac aaatgatgac
     3481 gaagcttgtc gagaagtact agaggatcat aatcagccat accacatttg tagaggtttt
     3541 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3601 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3661 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3721 caatgtatct tatcatgtct ggatctgatc actgatatcg cctaggagat ccgaaccaga
     3781 taagtgaaat ctagttccaa actattttgt catttttaat tttcgtatta gcttacgacg
     3841 ctacacccag ttcccatcta ttttgtcact cttccctaaa taatccttaa aaactccatt
     3901 tccacccctc ccagttccca actattttgt ccgcccacag cggggcattt ttcttcctgt
     3961 tatgttttta atcaaacatc ctgccaactc catgtgacaa accgtcatct tcggctactt
     4021 tttctctgtc acagaatgaa aatttttctg tcatctcttc gttattaatg tttgtaattg
     4081 actgaatatc aacgcttatt tgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
     4141 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4201 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4261 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4321 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4381 tttttcgccc tttgacgttg gagtccacgt tcttaatagt ggactcttgt tccaaactgg
     4441 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4501 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4561 attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4621 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4681 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4741 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4801 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4861 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4921 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4981 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     5041 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     5101 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     5161 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     5221 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     5281 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     5341 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5401 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5461 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5521 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5581 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5641 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5701 tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
