LOCUS       sequence-348397-        8854 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expression of the human active Cdk1/cyclin B1/Cks1 kinase 
            complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8854)
  AUTHORS   Yu J, Raia P, Ghent CM, Raisch T, Sadian Y, Cavadini S, Sabale PM, 
            Barford D, Raunser S, Morgan DO, Boland A
  TITLE     Structural basis of human separase regulation by securin and 
            CDK1-cyclin B1.
  JOURNAL   Nature. 2021 Aug;596(7870):138-142. doi: 10.1038/s41586-021-03764-0.
            Epub 2021 Jul 21.
  PUBMED    34290405
REFERENCE   2  (bases 1 to 8854)
  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: 
            "10.1038/s41586-021-03764-0"; journalName: "Nature"; date: 
            "2021-08"; volume: "596"; issue: "7870"; pages: "138-142"
COMMENT     Sequence Label: pF1-Hs Cdk1/cyclin B1/Cks1
FEATURES             Location/Qualifiers
     source          1..8854
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    116..250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  279..444
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      628..1082
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(715..734)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     924..945
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1108..1212
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1213..1281,1282..2073)
                     /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(1431..1450)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2244..2832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2733..2752
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2986..3003
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  3137..3361
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3428..3961)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4150..4155,4156..4172,4173..4178))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4695..4743
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(4824..4847)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(4891..4914)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(4951..4974)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(4975..4995)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(6310..6419)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        6464..6555
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             7462..7485
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             7486..7509
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     polyA_signal    7640..7774
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    7963..8011
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(8092..8115)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(8159..8182)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(8219..8242)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(8243..8263)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(8519..8628)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        8673..8764
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             8792..8815
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             8816..8839
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
ORIGIN
        1 gtactagagg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
       61 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
      121 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
      181 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
      241 tgtctggatc tgatcactgc ttgagcctag gagatccgaa ccagataagt gaaatctagt
      301 tccaaactat tttgtcattt ttaattttcg tattagctta cgacgctaca cccagttccc
      361 atctattttg tcactcttcc ctaaataatc cttaaaaact ccatttccac ccctcccagt
      421 tcccaactat tttgtccgcc cacagcgggg catttttctt cctgttatgt ttttaatcaa
      481 acatcctgcc aactccatgt gacaaaccgt catcttcggc tactttttct ctgtcacaga
      541 atgaaaattt ttctgtcatc tcttcgttat taatgtttgt aattgactga atatcaacgc
      601 ttatttgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
      661 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
      721 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
      781 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
      841 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
      901 cgttggagtc cacgttctta atagtggact cttgttccaa actggaacaa cactcaaccc
      961 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1021 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     1081 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     1141 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     1201 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     1261 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     1321 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     1381 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     1441 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     1501 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     1561 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     1621 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     1681 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     1741 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     1801 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     1861 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     1921 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     1981 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2041 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     2101 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     2161 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     2221 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     2281 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     2341 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     2401 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     2461 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     2521 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     2581 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     2641 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     2701 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     2761 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     2821 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     2881 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     2941 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     3001 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     3061 caccgcatag accagccgcg taacctggca aaatcggtta cggttgagta ataaatggat
     3121 gccctgcgta agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa atagatctaa
     3181 actatgacaa taaagtctta aactagacag aatagttgta aactgaaatc agtccagtta
     3241 tgctgtgaaa aagcatactg gacttttgtt atggctaaag caaactcttc attttctgaa
     3301 gtgcaaattg cccgtcgtat taaagagggg cgtggccaag ggcatggtaa agactatatt
     3361 cgcggcgttg tgacaattta ccgaacaact ccgcggccgg gaagccgatc tcggcttgaa
     3421 cgaattgtta ggtggcggta cttgggtcga tatcaaagtg catcacttct tcccgtatgc
     3481 ccaactttgt atagagagcc actgcgggat cgtcaccgta atctgcttgc acgtagatca
     3541 cataagcacc aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc
     3601 cctgcctccg gtgctcgccg gagactgcga gatcatagat atagatctca ctacgcggct
     3661 gctcaaactt gggcagaacg taagccgcga gagcgccaac aaccgcttct tggtcgaagg
     3721 cagcaagcgc gatgaatgtc ttactacgga gcaagttccc gaggtaatcg gagtccggct
     3781 gatgttggga gtaggtggct acgtctccga actcacgacc gaaaagatca agagcagccc
     3841 gcatggattt gacttggtca gggccgagcc tacatgtgcg aatgatgccc atacttgagc
     3901 cacctaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctgcgtaaca
     3961 tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg ctgcttggat
     4021 gcccgaggca tagactgtac aaaaaaacag tcataacaag ccatgaaaac cgccactgcg
     4081 ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg catacgctac
     4141 ttgcattaca gtttacgaac cgaacaggct tatgtcaact gggttcgtgc cttcatccgt
     4201 ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg aagtcgaggc atttctgtcc
     4261 tggctggcga acgagcgcaa ggtttcggtc tccacgcatc gtcaggcatt ggcggccttg
     4321 ctgttcttct acggcaaggt gctgtgcacg gatctgccct tgcttcagga gatcggtaga
     4381 cctcggccgt cgcggcgctt gccggtggtg ctgaccccgg atgaagtggt tcgcatcctc
     4441 ggttttctgg aaggcgagca tcgtttgttc gcccaggact ctagctatag ttctagtggt
     4501 tggctacagc tttgtttaaa caaagctggc tatggcaggg cttgccgccc cgacgttggc
     4561 tgcgagccct gggccttcac ccgaacttgg gggttggggt ggggaaaagg aagaaacgcg
     4621 ggcgtattgg tcccaatggg gtctcggtgg ggtatcgaca gagtgccagc cctgggaccg
     4681 aaccccgcgt ttatgaacaa acgacccaac acccgtgcgt tttattctgt ctttttattg
     4741 ccgtcatagc gcgggttcct tccggtattg tctccttccg tgtttcagtt agcctccccc
     4801 atctcccggt accgcatgct ttatttttca aattgcggat ggctccatgg aatgctggat
     4861 ttggtaccgc atgctatgca tcagctgtta cttctcgaac tgagggtgag accaagaacc
     4921 tccgccgcta cctccgccgg atccgccacc tttttcaaat tgcggatggc tccatgactg
     4981 gaagtacagg ttttcctcag ctagcgatga caccttagcc acagccttag ccaagtcctg
     5041 caccagagca gagttcagct ggggcagagt ggagatcttg gcgtgcttgg aggtggcgta
     5101 tttgttcttc acagtcatgt gcttggtcag gccctggttg accatgacca cgttcttggc
     5161 caggtgctgc atcacaggca gcagggattc ctcggtgtag gacaggtagt gctgcagagt
     5221 aggggtccac tcgccgttgt ccaggatctt gagagccagg cagaaagcac cagcagcgat
     5281 ctgagaaggt gggaagtgca ccatgtcgta gtccagcatg gtcagttcca tcaggtactt
     5341 agccagggtg tgctgttcca cgtccacctc tccgatcttg gaagcacgac gcaggaagtg
     5401 cagaggcaaa ggtctaccca ggccgaagtt cagagcgcgc aggatcttca tttccatctg
     5461 acggatctgg tgcttggtgt aggtgttgtc ggtcacgaaa gcgaagtcgc cgatctcagg
     5521 agggtacatt tcctcgtact tagaggcaat gaacatagcg gtgacaccca cgagctgcag
     5581 cattttcttg ggcacgcagt tgttctgcat gaagcggtcg atgatggaca cggtcatgta
     5641 catagtctct tgcagcagac ggaacttcat ctgcacctgg accagccagt cgatcaggat
     5701 agcacgcatg ttgccggtca cttcacgacc cagcaggtac ttgggacgca cagcctgttc
     5761 ctcttccagt tgacgcaggt aagcgtagat gtccttcacg tactcggagc acaggttagg
     5821 gtcagcaccg tcctcagcgt ccacgtcgtt agcagccagg atcacgtcgg agaaagcctg
     5881 gcacaggtcc tcttcagctg gagcgcaacc ggaggtttcc ataggtgagg gtgaagcggt
     5941 gtccaccagg ataggctcag gggacagctt ttcctctttc acaggaccag gttctggctc
     6001 tggttcaggc tcaggcacag gttcagacac aggcacgggc accaacatgg gcaccttctc
     6061 gagaggcttg ggcagcttct tgtcgatcac tttgccggta gcggagggct tagcctcttt
     6121 cttcatgggc atcttagcct gcagctgctc ggacaccttg tttccgatgt ctcccagagc
     6181 ggtacgagga cgcagtccag gcttagaagt agcagcagga gcggtgggca cacgcttagc
     6241 accagccatg ttgatcttcg ccttgttctc agcgttgatc ttggagttac gggtcacacg
     6301 cagagccatg attgtaaata aaatgtaatt tacagtatag tattttaatt aatatacaaa
     6361 tgatttgata ataattctta tttaactata atatattgtg ttgggttgaa ttaaaggtcc
     6421 gtatactagt atcgattcgc gacctactcc ggaatattaa tagatcatgg agataattaa
     6481 aatgataacc atctcgcaaa taaataagta ttttactgtt ttcgtaacag ttttgtaata
     6541 aaaaaaccta taaatatgga agattacacc aagatcgaga agatcggcga gggcacctac
     6601 ggtgtcgtgt acaagggtcg tcacaagacc accggacagg tggtggctat gaagaagatc
     6661 cgtctggaat ccgaggaaga gggcgtcccc tctaccgcta tccgtgaaat ctccctgctg
     6721 aaagagctgc gtcaccccaa catcgtgtcc ctgcaagacg tgctgatgca ggactcccgt
     6781 ctgtacctga tcttcgagtt cctgtctatg gacctgaaga agtacctgga ctctatccct
     6841 cctggccagt acatggactc cagcctggtc aagtcctacc tgtaccagat cctgcagggt
     6901 atcgtgttct gccactctcg tcgtgtgctg caccgtgacc tgaagcctca gaacctgctg
     6961 atcgacgaca agggcaccat caagctggct gacttcggac tggctcgtgc tttcggtatc
     7021 cccatccgtg tgtacaccca cgaggtggtc actctgtggt atcgttctcc cgaggtgctg
     7081 ctgggttccg ctcgttactc tacccctgtg gacatctggt ccatcggcac catcttcgct
     7141 gagctggcta ccaagaaacc cctgttccac ggcgactccg agatcgacca actgttccgt
     7201 atcttccgtg ctctgggcac ccctaacaac gaagtgtggc ctgaggtgga atctctgcag
     7261 gactacaaga acactttccc caagtggaag cccggttctc tggcttccca cgtgaagaac
     7321 ctggacgaga acggactgga cctgctgtcc aagatgctca tctacgaccc cgctaagcgc
     7381 atctccggca agatggctct gaaccatcct tacttcaacg acctggacaa ccagatcaag
     7441 aagatgatcg cagctgaggc tctggaagtt ctgtttcagg gacctcacca tcatcaccat
     7501 caccatcact aagcttgtcg agaagtacta gaggatcata atcagccata ccacatttgt
     7561 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     7621 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     7681 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     7741 caaactcatc aatgtatctt atcatgtctg gatctgatca ctgcttgagc ctaggcgctc
     7801 tgtccaggtt caatcgcccg acgttggctg cgagccctgg gccttcaccc gaacttgggg
     7861 gttggggtgg ggaaaaggaa gaaacgcggg cgtattggtc ccaatggggt ctcggtgggg
     7921 tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt atgaacaaac gacccaacac
     7981 ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc
     8041 tccttccgtg tttcagttag cctcccccat ctcccggtac cgcatgcttt atttttcaaa
     8101 ttgcggatgg ctccatggaa tgctggattt ggtaccgcat gctatgcatc agctgttact
     8161 tctcgaactg agggtgagac caagaacctc cgccgctacc tccgccggat ccgccacctt
     8221 tttcaaattg cggatggctc catgactgga agtacaggtt ttcctcagct agcgatgatt
     8281 acttcttggg cttcttaggc agaggacgac ggaacagcag gatatgaggc tcaggctcgt
     8341 ggatcatgta gtgcacccag ccttgagact gctggacacc caggttacgc cactcggact
     8401 cggacatcag gtgagtcttg ggcaccagct tagcgatatc cttgggcagc atcacgtgac
     8461 ggtactcgaa ttcctcgtcg tcgtacttat cggagtagta gatctgcttg tgggacatga
     8521 ttgtaaataa aatgtaattt acagtatagt attttaatta atatacaaat gatttgataa
     8581 taattcttat ttaactataa tatattgtgt tgggttgaat taaaggtccg tatactagta
     8641 tcgattcgcg acctactccg gaatattaat agatcatgga gataattaaa atgataacca
     8701 tctcgcaaat aaataagtat tttactgttt tcgtaacagt tttgtaataa aaaaacctat
     8761 aaatatccct ctctccatcg cagctgaggc tctggaagtt ctgtttcagg gacctcacca
     8821 tcatcaccat caccatcact aagcttgtcg agaa
//
