LOCUS       sequence-348398-        7524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Co-expression of the human active Cdk1/cyclin B1 kinase complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7524)
  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 7524)
  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
FEATURES             Location/Qualifiers
     source          1..7524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      95..549
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(182..201)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     391..412
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        575..679
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(680..748,749..1540)
                     /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(898..917)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1711..2299
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2200..2219
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2453..2470
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2604..2828
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2895..3428)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3617..3622,3623..3639,3640..3645))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    4162..4210
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4235..4254
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4291..4314)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(4358..4381)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(4418..4441)
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             complement(4442..4462)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        complement(5777..5886)
                     /label=p10 promoter
                     /note="baculovirus promoter for expression in insect cells"
     promoter        5931..6022
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     5947..5964
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             6929..6952
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6953..6976
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     polyA_signal    7107..7241
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7144..7163)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7198..7217
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  7270..7435
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 tcacagaatg aaaatttttc tgtcatctct tcgttattaa tgtttgtaat tgactgaata
       61 tcaacgctta tttgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
      121 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
      181 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
      241 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
      301 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
      361 cctttgacgt tggagtccac gttcttaata gtggactctt gttccaaact ggaacaacac
      421 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
      481 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
      541 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
      601 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
      661 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
      721 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
      781 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
      841 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
      901 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
      961 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     1021 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     1081 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     1141 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     1201 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     1261 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     1321 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     1381 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     1441 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     1501 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     1561 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     1621 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     1681 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     1741 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     1801 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     1861 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     1921 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     1981 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2041 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2101 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2161 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2221 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2281 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2341 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2401 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2461 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     2521 tatttcacac cgcatagacc agccgcgtaa cctggcaaaa tcggttacgg ttgagtaata
     2581 aatggatgcc ctgcgtaagc gggtgtgggc ggacaataaa gtcttaaact gaacaaaata
     2641 gatctaaact atgacaataa agtcttaaac tagacagaat agttgtaaac tgaaatcagt
     2701 ccagttatgc tgtgaaaaag catactggac ttttgttatg gctaaagcaa actcttcatt
     2761 ttctgaagtg caaattgccc gtcgtattaa agaggggcgt ggccaagggc atggtaaaga
     2821 ctatattcgc ggcgttgtga caatttaccg aacaactccg cggccgggaa gccgatctcg
     2881 gcttgaacga attgttaggt ggcggtactt gggtcgatat caaagtgcat cacttcttcc
     2941 cgtatgccca actttgtata gagagccact gcgggatcgt caccgtaatc tgcttgcacg
     3001 tagatcacat aagcaccaag cgcgttggcc tcatgcttga ggagattgat gagcgcggtg
     3061 gcaatgccct gcctccggtg ctcgccggag actgcgagat catagatata gatctcacta
     3121 cgcggctgct caaacttggg cagaacgtaa gccgcgagag cgccaacaac cgcttcttgg
     3181 tcgaaggcag caagcgcgat gaatgtctta ctacggagca agttcccgag gtaatcggag
     3241 tccggctgat gttgggagta ggtggctacg tctccgaact cacgaccgaa aagatcaaga
     3301 gcagcccgca tggatttgac ttggtcaggg ccgagcctac atgtgcgaat gatgcccata
     3361 cttgagccac ctaactttgt tttagggcga ctgccctgct gcgtaacatc gttgctgctg
     3421 cgtaacatcg ttgctgctcc ataacatcaa acatcgaccc acggcgtaac gcgcttgctg
     3481 cttggatgcc cgaggcatag actgtacaaa aaaacagtca taacaagcca tgaaaaccgc
     3541 cactgcgccg ttaccaccgc tgcgttcggt caaggttctg gaccagttgc gtgagcgcat
     3601 acgctacttg cattacagtt tacgaaccga acaggcttat gtcaactggg ttcgtgcctt
     3661 catccgtttc cacggtgtgc gtcacccggc aaccttgggc agcagcgaag tcgaggcatt
     3721 tctgtcctgg ctggcgaacg agcgcaaggt ttcggtctcc acgcatcgtc aggcattggc
     3781 ggccttgctg ttcttctacg gcaaggtgct gtgcacggat ctgcccttgc ttcaggagat
     3841 cggtagacct cggccgtcgc ggcgcttgcc ggtggtgctg accccggatg aagtggttcg
     3901 catcctcggt tttctggaag gcgagcatcg tttgttcgcc caggactcta gctatagttc
     3961 tagtggttgg ctacagcttt gtttaaacaa agctggctat ggcagggctt gccgccccga
     4021 cgttggctgc gagccctggg ccttcacccg aacttggggg ttggggtggg gaaaaggaag
     4081 aaacgcgggc gtattggtcc caatggggtc tcggtggggt atcgacagag tgccagccct
     4141 gggaccgaac cccgcgttta tgaacaaacg acccaacacc cgtgcgtttt attctgtctt
     4201 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc
     4261 ctcccccatc tcccggtacc gcatgcttta tttttcaaat tgcggatggc tccatggaat
     4321 gctggatttg gtaccgcatg ctatgcatca gctgttactt ctcgaactga gggtgagacc
     4381 aagaacctcc gccgctacct ccgccggatc cgccaccttt ttcaaattgc ggatggctcc
     4441 atgactggaa gtacaggttt tcctcagcta gcgatgacac cttagccaca gccttagcca
     4501 agtcctgcac cagagcagag ttcagctggg gcagagtgga gatcttggcg tgcttggagg
     4561 tggcgtattt gttcttcaca gtcatgtgct tggtcaggcc ctggttgacc atgaccacgt
     4621 tcttggccag gtgctgcatc acaggcagca gggattcctc ggtgtaggac aggtagtgct
     4681 gcagagtagg ggtccactcg ccgttgtcca ggatcttgag agccaggcag aaagcaccag
     4741 cagcgatctg agaaggtggg aagtgcacca tgtcgtagtc cagcatggtc agttccatca
     4801 ggtacttagc cagggtgtgc tgttccacgt ccacctctcc gatcttggaa gcacgacgca
     4861 ggaagtgcag aggcaaaggt ctacccaggc cgaagttcag agcgcgcagg atcttcattt
     4921 ccatctgacg gatctggtgc ttggtgtagg tgttgtcggt cacgaaagcg aagtcgccga
     4981 tctcaggagg gtacatttcc tcgtacttag aggcaatgaa catagcggtg acacccacga
     5041 gctgcagcat tttcttgggc acgcagttgt tctgcatgaa gcggtcgatg atggacacgg
     5101 tcatgtacat agtctcttgc agcagacgga acttcatctg cacctggacc agccagtcga
     5161 tcaggatagc acgcatgttg ccggtcactt cacgacccag caggtacttg ggacgcacag
     5221 cctgttcctc ttccagttga cgcaggtaag cgtagatgtc cttcacgtac tcggagcaca
     5281 ggttagggtc agcaccgtcc tcagcgtcca cgtcgttagc agccaggatc acgtcggaga
     5341 aagcctggca caggtcctct tcagctggag cgcaaccgga ggtttccata ggtgagggtg
     5401 aagcggtgtc caccaggata ggctcagggg acagcttttc ctctttcaca ggaccaggtt
     5461 ctggctctgg ttcaggctca ggcacaggtt cagacacagg cacgggcacc aacatgggca
     5521 ccttctcgag aggcttgggc agcttcttgt cgatcacttt gccggtagcg gagggcttag
     5581 cctctttctt catgggcatc ttagcctgca gctgctcgga caccttgttt ccgatgtctc
     5641 ccagagcggt acgaggacgc agtccaggct tagaagtagc agcaggagcg gtgggcacac
     5701 gcttagcacc agccatgttg atcttcgcct tgttctcagc gttgatcttg gagttacggg
     5761 tcacacgcag agccatgatt gtaaataaaa tgtaatttac agtatagtat tttaattaat
     5821 atacaaatga tttgataata attcttattt aactataata tattgtgttg ggttgaatta
     5881 aaggtccgta tactagtatc gattcgcgac ctactccgga atattaatag atcatggaga
     5941 taattaaaat gataaccatc tcgcaaataa ataagtattt tactgttttc gtaacagttt
     6001 tgtaataaaa aaacctataa atatggaaga ttacaccaag atcgagaaga tcggcgaggg
     6061 cacctacggt gtcgtgtaca agggtcgtca caagaccacc ggacaggtgg tggctatgaa
     6121 gaagatccgt ctggaatccg aggaagaggg cgtcccctct accgctatcc gtgaaatctc
     6181 cctgctgaaa gagctgcgtc accccaacat cgtgtccctg caagacgtgc tgatgcagga
     6241 ctcccgtctg tacctgatct tcgagttcct gtctatggac ctgaagaagt acctggactc
     6301 tatccctcct ggccagtaca tggactccag cctggtcaag tcctacctgt accagatcct
     6361 gcagggtatc gtgttctgcc actctcgtcg tgtgctgcac cgtgacctga agcctcagaa
     6421 cctgctgatc gacgacaagg gcaccatcaa gctggctgac ttcggactgg ctcgtgcttt
     6481 cggtatcccc atccgtgtgt acacccacga ggtggtcact ctgtggtatc gttctcccga
     6541 ggtgctgctg ggttccgctc gttactctac ccctgtggac atctggtcca tcggcaccat
     6601 cttcgctgag ctggctacca agaaacccct gttccacggc gactccgaga tcgaccaact
     6661 gttccgtatc ttccgtgctc tgggcacccc taacaacgaa gtgtggcctg aggtggaatc
     6721 tctgcaggac tacaagaaca ctttccccaa gtggaagccc ggttctctgg cttcccacgt
     6781 gaagaacctg gacgagaacg gactggacct gctgtccaag atgctcatct acgaccccgc
     6841 taagcgcatc tccggcaaga tggctctgaa ccatccttac ttcaacgacc tggacaacca
     6901 gatcaagaag atgatcgcag ctgaggctct ggaagttctg tttcagggac ctcaccatca
     6961 tcaccatcac catcactaag cttgtcgaga agtactagag gatcataatc agccatacca
     7021 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     7081 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     7141 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     7201 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctgatcactg cttgagccta
     7261 ggagatccga accagataag tgaaatctag ttccaaacta ttttgtcatt tttaattttc
     7321 gtattagctt acgacgctac acccagttcc catctatttt gtcactcttc cctaaataat
     7381 ccttaaaaac tccatttcca cccctcccag ttcccaacta ttttgtccgc ccacagcggg
     7441 gcatttttct tcctgttatg tttttaatca aacatcctgc caactccatg tgacaaaccg
     7501 tcatcttcgg ctactttttc tctg
//
