LOCUS       sequence-469841-        7871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7871)
  AUTHORS   Seibert M, Kruger M, Watson NA, Sen O, Daum JR, Slotman JA, Braun T,
            Houtsmuller AB, Gorbsky GJ, Jacob R, Kracht M, Higgins JMG, Schmitz 
            ML
  TITLE     CDK1-mediated phosphorylation at H2B serine 6 is required for 
            mitotic chromosome segregation.
  JOURNAL   J Cell Biol. 2019 Apr 1;218(4):1164-1181. doi: 
            10.1083/jcb.201806057. Epub 2019 Feb 14.
  PUBMED    30765437
REFERENCE   2  (bases 1 to 7871)
  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: 
            "10.1083/jcb.201806057"; journalName: "J Cell Biol"; date: 
            "2019-04-1- 1"; volume: "218"; issue: "4"; pages: "1164-1181"
COMMENT     Sequence Label: pEGFP-BUBR1
FEATURES             Location/Qualifiers
     source          1..7871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /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(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1379..1388
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4659..4780
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4696..4715)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4750..4769
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4787..5242)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4924..4945)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5136..5155
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5269..5373
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5375..5732
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5375..5566
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5398..5418)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5583..5718
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5767..6561
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5767..6561
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5821..5840)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6431..6450
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6749..6768)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6793..6840
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7169..7757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7658..7677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc
     1381 atggcggcgg tgaagaagga agggggtgct ctgagtgaag ccatgtccct ggagggagat
     1441 gaatgggaac tgagtaaaga aaatgtacaa cctttaaggc aagggcggat catgtccacg
     1501 cttcagggag cactggcaca agaatctgcc tgtaacaata ctcttcagca gcagaaacgg
     1561 gcatttgaat atgaaattcg attttacact ggaaatgacc ctctggatgt ttgggatagg
     1621 tatatcagct ggacagagca gaactatcct caaggtggga aggagagtaa tatgtcaacg
     1681 ttattagaaa gagctgtaga agcactacaa ggagaaaaac gatattatag tgatcctcga
     1741 tttctcaatc tctggcttaa attagggcgt ttatgcaatg agcctttgga tatgtacagt
     1801 tacttgcaca accaagggat tggtgtttca cttgctcagt tctatatctc atgggcagaa
     1861 gaatatgaag ctagagaaaa ctttaggaaa gcagatgcga tatttcagga agggattcaa
     1921 cagaaggctg aaccactaga aagactacag tcccagcacc gacaattcca agctcgagtg
     1981 tctcggcaaa ctctgttggc acttgagaaa gaagaagagg aggaagtttt tgagtcttct
     2041 gtaccacaac gaagcacact agctgaacta aagagcaaag ggaaaaagac agcaagagct
     2101 ccaatcatcc gtgtaggagg tgctctcaag gctccaagcc agaacagagg actccaaaat
     2161 ccatttcctc aacagatgca aaataatagt agaattactg tttttgatga aaatgctgat
     2221 gaggcttcta cagcagagtt gtctaagcct acagtccagc catggatagc accccccatg
     2281 cccagggcca aagagaatga gctgcaagca ggcccttgga acacaggcag gtccttggaa
     2341 cacaggcctc gtggcaatac agcttcactg atagctgtac ccgctgtgct tcccagtttc
     2401 actccatatg tggaagagac tgcacaacag ccagttatga caccatgtaa aattgaacct
     2461 agtataaacc acatcctaag caccagaaag cctggaaagg aagaaggaga ttctctacaa
     2521 agggttcaga gccatcagca agcgtctgag gagaagaaag agaagatgat gtattgtaag
     2581 gagaagattt atgcaggagt aggggaattc tcctttgaag aaattcgggc tgaagttttc
     2641 cggaagaaat taaaagagca aagggaagcc gagctattga ccagtgcaga gaagagagca
     2701 gaaatgcaga aacagattga agagatggag aagaagctaa aagaaatcca aactactcag
     2761 caagaaagaa caggtgatca gcaagaagag acgatgccta caaaggagac aactaaactg
     2821 caaattgctt ccgagtctca gaaaatacca ggaatgactc tatccagttc tgtttgtcaa
     2881 gtaaactgtt gtgccagaga aacttcactt gcggagaaca tttggcagga acaacctcat
     2941 tctaaaggtc ccagtgtacc tttctccatt tttgatgagt ttcttctttc agaaaagaag
     3001 aataaaagtc ctcctgcaga tcccccacga gttttagctc aacgaagacc ccttgcagtt
     3061 ctcaaaacct cagaaagcat cacctcaaat gaagatgtgt ctccagatgt ttgtgatgaa
     3121 tttacaggaa ttgaaccctt gagcgaggat gccattatca caggcttcag aaatgtaaca
     3181 atttgtccta acccagaaga cacttgtgac tttgccagag cagctcgttt tgtatccact
     3241 ccttttcatg agataatgtc cttgaaggat ctcccttctg atcctgagag actgttaccg
     3301 gaagaagatc tagatgtaaa gacctctgag gaccagcaga cagcttgtgg cactatctac
     3361 agtcagactc tcagcatcaa gaagctgagc ccaattattg aagacagtcg tgaagccaca
     3421 cactcctctg gcttctctgg ttcttctgcc tcggttgcaa gcacctcctc catcaaatgt
     3481 cttcaaattc ctgagaaact agaacttact aatgagactt cagaaaaccc tactcagtca
     3541 ccatggtgtt cacagtatcg cagacagcta ctgaagtccc taccagagtt aagtgcctct
     3601 gcagagttgt gtatagaaga cagaccaatg cctaagttgg aaattgagaa ggaaattgaa
     3661 ttaggtaatg aggattactg cattaaacga gaatacctaa tatgtgaaga ttacaagtta
     3721 ttctgggtgg cgccaagaaa ctctgcagaa ttaacagtaa taaaggtatc ttctcaacct
     3781 gtcccatggg acttttatat caacctcaag ttaaaggaac gtttaaatga agattttgat
     3841 catttttgca gctgttatca atatcaagat ggctgtattg tttggcacca atatataaac
     3901 tgcttcaccc ttcaggatct tctccaacac agtgaatata ttacccatga aataacagtg
     3961 ttgattattt ataacctttt gacaatagtg gagatgctac acaaagcaga aatagtccat
     4021 ggtgacttga gtccaaggtg tctgattctc agaaacagaa tccacgatcc ctatgattgt
     4081 aacaagaaca atcaagcttt gaagatagtg gacttttcct acagtgttga ccttagggtg
     4141 cagctggatg tttttaccct cagcggcttt cggactgtac agatcctgga aggacaaaag
     4201 atcctggcta actgttcttc tccctaccag gtagacctgt ttggtatagc agatttagca
     4261 catttactat tgttcaagga acacctacag gtcttctggg atgggtcctt ctggaaactt
     4321 agccaaaata tttctgagct aaaagatggt gaattgtgga ataaattctt tgtgcggatt
     4381 ctgaatgcca atgatgaggc cacagtgtct gttcttgggg agcttgcagc agaaatgaat
     4441 ggggtttttg acactacatt ccaaagtcac ctgaacaaag ccttatggaa ggtagggaag
     4501 ttaactagtc ctggggcttt gctctttcag gatccaccgg atctagataa ctgatcataa
     4561 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     4621 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     4681 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     4741 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     4801 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     4861 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     4921 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     4981 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     5041 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     5101 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     5161 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     5221 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5281 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5341 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     5401 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5461 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5521 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5581 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5641 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     5701 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     5761 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     5821 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     5881 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     5941 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     6001 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     6061 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     6121 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     6181 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     6241 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     6301 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     6361 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     6421 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     6481 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     6541 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     6601 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     6661 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     6721 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     6781 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     6841 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     6901 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     6961 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     7021 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     7081 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7141 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7201 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7261 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7321 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7381 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7441 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7501 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7561 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7621 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7681 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7741 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7801 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     7861 ccgccatgca t
//
