LOCUS       sequence-313552-        6016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6016)
  AUTHORS   Gavet O, Pines J
  TITLE     Progressive activation of CyclinB1-Cdk1 coordinates entry to 
            mitosis.
  JOURNAL   Dev Cell. 2010 Apr 20. 18(4):533-43.
  PUBMED    20412769
REFERENCE   2  (bases 1 to 6016)
  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: "Dev Cell. 
            2010 Apr 20. 18(4):533-43."
COMMENT     Sequence Label: p-cyclinB1-mcherry (1958)
FEATURES             Location/Qualifiers
     source          1..6016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1906..1915
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1912..2622
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2057..2075)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2325..2344
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2744..2865
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2781..2800)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2835..2854
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2872..3327)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3009..3030)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3221..3240
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3354..3458
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3460..3817
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3460..3651
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3483..3503)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3668..3803
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3852..4646
                     /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             3852..4646
                     /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(3906..3925)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4516..4535
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4834..4853)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4878..4925
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5254..5842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5743..5762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttata tctgaggagg acctttccat ggcgctccga
      601 gccaccagga actcgaaaat taatgctgaa aataaggcga agatcaacat ggcaggcgca
      661 aagcgcgttc ctacggcccc tgctgcaacc tccaagcccg gactgaggcc aagaacagct
      721 cttggggaca ttggtaacaa agtcagtgaa caactgcagg ccaaaatgcc tatgaagaag
      781 gaagcaaaac cttcagctac tggaaaagtc attgataaaa aactaccaaa acctcttgaa
      841 aaggtaccta tgctggtgcc agtgccagtg tctgagccag tgccagagcc agaacctgag
      901 ccagaacctg agcctgttaa agaagaaaaa ctttcgcctg agcctatttt ggttgatact
      961 gcctctccaa gcccaatgga aacatctgga tgtgcccctg cagaagaaga cctgtgtcag
     1021 gctttctctg atgtaattct tgcagtaaat gatgtggatg cagaagatgg agctgatcca
     1081 aacctttgta gtgaatatgt gaaagatatt tatgcttatc tgagacaact tgaggaagag
     1141 caagcagtca gaccaaaata cctactgggt cgggaagtca ctggaaacat gagagccatc
     1201 ctaattgact ggctagtaca ggttcaaatg aaattcaggt tgttgcagga gaccatgtac
     1261 atgactgtct ccattattga tcggttcatg cagaataatt gtgtgcccaa gaagatgctg
     1321 cagctggttg gtgtcactgc catgtttatt gcaagcaaat atgaagaaat gtaccctcca
     1381 gaaattggtg actttgcttt tgtgactgac aacacttata ctaagcacca aatcagacag
     1441 atggaaatga agattctaag agctttaaac tttggtctgg gtcggcctct acctttgcac
     1501 ttccttcgga gagcatctaa gattggagag gttgatgtcg agcaacatac tttggccaaa
     1561 tacctgatgg aactaactat gttggactat gacatggtgc actttcctcc ttctcaaatt
     1621 gcagcaggag ctttttgctt agcactgaaa attctggata atggtgaatg gacaccaact
     1681 ctacaacatt acctgtcata tactgaagaa tctcttcttc cagttatgca gcacctggct
     1741 aagaatgtag tcatggtaaa tcaaggactt acaaagcaca tgactgtcaa gaacaagtat
     1801 gccacatcga agcatgctaa gatcagcact ctaccacagc tgaattctgc actagttcaa
     1861 gatttagcca aggctgtggc aaaggttcta gaggatccac cggtcgccac catggtgagc
     1921 aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa ggtgcacatg
     1981 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     2041 gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc cttcgcctgg
     2101 gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca ccccgccgac
     2161 atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     2221 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcgagttc
     2281 atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt aatgcagaag
     2341 aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg cgccctgaag
     2401 ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc tgaggtcaag
     2461 accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt caacatcaag
     2521 ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga acgcgccgag
     2581 ggccgccact ccaccggcgg catggacgag ctgtacaagt aagcggccgc gactctagat
     2641 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2701 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2761 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2821 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa
     2881 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2941 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3001 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3061 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3121 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3181 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3241 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3301 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3361 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3421 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3481 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3541 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3601 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3661 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3721 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3781 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3841 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3901 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3961 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4021 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4081 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4141 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4201 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4261 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4321 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4381 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4441 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4501 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4561 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4621 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4681 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4741 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4801 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4861 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     4921 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4981 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     5041 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     5101 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5161 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5221 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5281 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5341 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5401 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5461 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5521 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5581 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5641 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5701 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5761 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5821 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5881 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5941 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     6001 catatatgga gttccg
//
