LOCUS Clover-Geminin(1 7475 bp DNA circular SYN 13-MAY-2021 DEFINITION Fluorescent probe for M/G1 transition. ACCESSION . VERSION . KEYWORDS Clover-Geminin(1-110). SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7475) AUTHORS Bajar BT, Lam AJ, Badiee RK, Oh YH, Chu J, Zhou XX, Kim N, Kim BB, Chung M, Yablonovitch AL, Cruz BF, Kulalert K, Tao JJ, Meyer T, Su XD, Lin MZ TITLE Fluorescent indicators for simultaneous reporting of all four cell cycle phases. JOURNAL Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4045. PUBMED 27798610 REFERENCE 2 (bases 1 to 7475) TITLE Direct Submission REFERENCE 3 (bases 1 to 7475) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Methods. 2016 Oct 31. doi: 10.1038/nmeth.4045." COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..7475 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 63..366 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 368..566 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 584..764 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 811..936 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1435..1668 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1853..1897 /label=gp41 peptide /note="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" CDS 2046..2087 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" misc_feature 2195..2312 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2371..2684 /label=U6 promoter /note="RNA polymerase III promoter for mouse U6 snRNA (Das et al., 1988)" primer_bind 2701..2717 /label=KS primer /note="common sequencing primer, one of multiple similar variants" protein_bind 2759..2792 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." enhancer 2908..3211 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3212..3415 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" CDS 3447..4163 /label=Clover /note="bright green-yellow fluorescent protein derived from GFP (Lam et al., 2012)" CDS 4194..4523 /codon_start=1 /product="degron consisting of residues 1-110 of human Geminin (Zielke and Edgar, 2015)" /label=Gem1 (1-110) /translation="MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELS AGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAE KRRKAL" protein_bind complement(4562..4595) /label=loxP /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." misc_feature 4651..5239 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5242..5258) /label=KS primer /note="common sequencing primer, one of multiple similar variants" LTR 5481..5661 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(5723..6311) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6485..7342) /label=AmpR /note="beta-lactamase" promoter complement(7343..7447) /label=AmpR promoter ORIGIN 1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc 61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca 121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt 181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg 241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag 301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt 361 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg 421 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa 481 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt 541 gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc tctggttaga 601 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 661 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 721 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg 781 gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa 841 gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc 901 ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga 961 tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat 1021 atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca 1081 tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa 1141 gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag 1201 ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc 1261 accgcacagc aagcggccgg ccgcgctgat cttcagacct ggaggaggag atatgaggga 1321 caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat taggagtagc 1381 acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg gaataggagc 1441 tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt caatgacgct 1501 gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca atttgctgag 1561 ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca agcagctcca 1621 ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg ggatttgggg 1681 ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt ggagtaataa 1741 atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag aaattaacaa 1801 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga 1861 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa 1921 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat 1981 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt 2041 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg 2101 tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac tgcgtgcgcc 2161 aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg gggattgggg 2221 ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa actaaagaat 2281 tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac agcagagatc 2341 cagtttggtt agtaccgggc ccgctctaga gatccgacgc cgccatctct aggcccgcgc 2401 cggccccctc gcacagactt gtgggagaag ctcggctact cccctgcccc ggttaatttg 2461 catataatat ttcctagtaa ctatagaggc ttaatgtgcg ataaaagaca gataatctgt 2521 tctttttaat actagctaca ttttacatga taggcttgga tttctataag agatacaaat 2581 actaaattat tattttaaaa aacagcacaa aaggaaactc accctaactg taaagtaatt 2641 gtgtgttttg agactataaa tatcccttgg agaaaagcct tgttaacgcg cggtgaccct 2701 cgaggtcgac ggtatcgata agctcgcttc acgagattcc agcaggtcga gggacctaat 2761 aacttcgtat agcatacatt atacgaagtt atattaaggg ttccaagctt aagcggccgc 2821 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt 2881 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg 2941 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac 3001 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt 3061 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa 3121 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta 3181 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg 3241 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg 3301 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc 3361 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt 3421 agtgaaccgt cagatccgct agcaccatgg tgagcaaggg cgaggagctg ttcaccgggg 3481 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtccgcg 3541 gcgagggcga gggcgatgcc accaacggca agctgaccct gaagttcatc tgcaccaccg 3601 gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc gtggcctgct 3661 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag 3721 gctacgtcca ggagcgcacc atctctttca aggacgacgg tacctacaag acccgcgccg 3781 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca 3841 aggaggacgg caacatcctg gggcacaagc tggagtacaa cttcaacagc cacaacgtct 3901 atatcacggc cgacaagcag aagaacggca tcaaggctaa cttcaagatc cgccacaacg 3961 ttgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg 4021 gccccgtgct gctgcccgac aaccactacc tgagccatca gtccgccctg agcaaagacc 4081 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggattacac 4141 atggcatgga cgagctgtac aaggatatcc atgcacactg gcggccgctc gagatgaatc 4201 ccagtatgaa gcagaaacaa gaagaaatca aagagaatat aaagaatagt tctgtcccaa 4261 gaagaactct gaagatgatt cagccttctg catctggatc tcttgttgga agagaaaatg 4321 agctgtccgc aggcttgtcc aaaaggaaac atcggaatga ccacttaaca tctacaactt 4381 ccagccctgg ggttattgtc ccagaatcta gtgaaaataa aaatcttgga ggagtcaccc 4441 aggagtcatt tgatcttatg attaaagaaa atccatcctc tcagtattgg aaggaagtgg 4501 cagaaaaacg gagaaaggcg ctgtaagttt aaaccctgca gggaattcgt cgagggacct 4561 aataacttcg tatagcatac attatacgaa gttatacatg tttaagggtt ccggttccac 4621 taggtacaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa aatttgtgaa 4681 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta 4741 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa 4801 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg 4861 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc 4921 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc 4981 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg 5041 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg 5101 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg 5161 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc 5221 ctttgggccg cctccccgca tcgataccgt cgacctcgat cgagacctag aaaaacatgg 5281 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca 5341 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta 5401 caaggcagct gacgcgtgat tttaaaagaa aaggggggac tggaagggct aattcactcc 5461 cactgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct 5521 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 5581 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 5641 cagtgtggaa aatctctagc agcatgtgag caaaaggcca gcaaaaggcc aggaaccgta 5701 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 5761 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 5821 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 5881 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 5941 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 6001 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 6061 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 6121 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 6181 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 6241 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 6301 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 6361 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 6421 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 6481 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 6541 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 6601 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa 6661 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc 6721 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca 6781 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat 6841 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag 6901 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac 6961 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt 7021 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt 7081 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc 7141 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat 7201 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca 7261 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga 7321 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg 7381 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 7441 ttccgcgcac atttccccga aaagtgccac ctgac //