LOCUS       sequence-474718-        6008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses a non-degradable cyclin B1 in fusion with Venus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6008)
  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 6008)
  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: Cyclin B1-Venus ND
FEATURES             Location/Qualifiers
     source          1..6008
                     /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      1948..1957
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1954..1956,1957..1959,1960..2673)
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1999..2020)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2260..2279)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2607..2628
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2796..2917
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2833..2852)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2887..2906
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2924..3379)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3061..3082)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3273..3292
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3406..3510
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3512..3869
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3512..3703
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3535..3555)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3720..3855
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3904..4698
                     /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             3904..4698
                     /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(3958..3977)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4568..4587
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4886..4905)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4930..4977
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5306..5894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5795..5814
                     /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 ccggactcag atctcgagct caagcttgga atggcgctcc gagtcaccag gaactcgaaa
      661 attaatgctg aaaataaggc gaagatcaac atggcaggcg caaagcgcgt tcctacggcc
      721 cctgctgcaa cctccaagcc cggactgagg ccagcaacag ctgctgggga cattggtaac
      781 aaagtcagtg aacaactgca ggccaaaatg cctatgaaga aggaagcaaa accttcagct
      841 actggaaaag tcattgataa aaaactacca aaacctcttg aaaaggtacc tatgctggtg
      901 ccagtgccag tgtctgagcc agtgccagag ccagaacctg agccagaacc tgagcctgtt
      961 aaagaagaaa aactttcgcc tgagcctatt ttggttgata ctgcctctcc aagcccaatg
     1021 gaaacatctg gatgtgcccc tgcagaagaa gacctgtgtc aggctttctc tgatgtaatt
     1081 cttgcagtaa atgatgtgga tgcagaagat ggagctgatc caaacctttg tagtgaatat
     1141 gtgaaagata tttatgctta tctgagacaa cttgaggaag agcaagcagt cagaccaaaa
     1201 tacctactgg gtcgggaagt cactggaaac atgagagcca tcctaattga ctggctagta
     1261 caggttcaaa tgaaattcag gttgttgcag gagaccatgt acatgactgt ctccattatt
     1321 gatcggttca tgcagaataa ttgtgtgccc aagaagatgc tgcagctggt tggtgtcact
     1381 gccatgttta ttgcaagcaa atatgaagaa atgtaccctc cagaaattgg tgactttgct
     1441 tttgtgactg acaacactta tactaagcac caaatcagac agatggaaat gaagattcta
     1501 agagctttaa actttggtct gggtcggcct ctacctttgc acttccttcg gagagcatct
     1561 aagattggag aggttgatgt cgagcaacat actttggcca aatacctgat ggaactaact
     1621 atgttggact atgacatggt gcactttcct ccttctcaaa ttgcagcagg agctttttgc
     1681 ttagcactga aaattctgga taatggtgaa tggacaccaa ctctacaaca ttacctgtca
     1741 tatactgaag aatctcttct tccagttatg cagcacctgg ctaagaatgt agtcatggta
     1801 aatcaaggac ttacaaagca catgactgtc aagaacaagt atgccacatc gaagcatgct
     1861 aagatcagca ctctaccaca gctgaattct gcactagttc aagatttagc caaggctgtg
     1921 gcaaaggttc tagaggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc
     1981 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     2041 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gctgatctgc
     2101 accaccggca agctgcccgt gccctggccc accctcgtga ccactctggg ctacggccta
     2161 cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     2221 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     2281 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     2341 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     2401 aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc
     2461 cacaacatcg aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc
     2521 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgagc
     2581 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     2641 atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag atcataatca
     2701 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2761 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2821 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2881 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     2941 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3001 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3061 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3121 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3181 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3241 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3301 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3361 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3421 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3481 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3541 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3601 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3661 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3721 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3781 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3841 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3901 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3961 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4021 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4081 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4141 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4201 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4261 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4321 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4381 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4441 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4501 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4561 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4621 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4681 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4741 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4801 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4861 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4921 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4981 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     5041 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     5101 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     5161 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5221 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5281 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5341 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5401 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5461 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5521 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5581 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5641 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5701 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5761 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5821 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5881 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5941 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     6001 ccatgcat
//
