LOCUS Exported 7426 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Fluorescent probe for S/G2 cell cycle transition. ACCESSION . VERSION . KEYWORDS mKO2-SLBP(18-126) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7426) 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 7426) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7426 /organism="synthetic DNA construct" /mol_type="other DNA" 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 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" 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 2603..2623 /label=mU6-F /note="Mouse U6 promoter, forward primer" 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 3451..4101 /codon_start=1 /product="monomeric Kusabira-Orange 2 fluorescent protein" /label=mKO2 /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH" protein_bind 4513..4546 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 4602..5190 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4655..4675) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5073..5084) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(5193..5209) /label=KS primer /note="common sequencing primer, one of multiple similar variants" LTR 5432..5612 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(5674..6262) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5754..5773) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6433..7293) /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 7056..7075 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7294..7398) /gene="bla" /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 agcaccaggt atggtgagcg tgatcaagcc cgagatgaag 3481 atgaggtact acatggacgg ctccgtcaat gggcatgagt tcacaatcga gggtgagggc 3541 acaggcagac cttacgaggg acatcaggag atgacactgc gcgtcacaat ggccgagggc 3601 gggccaatgc ctttcgcctt cgacctggtg tcccacgtgt tctgttacgg ccacagagtt 3661 tttaccaagt acccagaaga gatcccagac tatttcaagc aggcctttcc tgagggcctg 3721 tcctgggaga ggtccctgga gttcgaggac ggcggctccg cctccgtgag cgcccacatc 3781 agcctgaggg gcaacacctt ctaccacaag tccaagttca ccggcgtgaa cttccccgcc 3841 gacggcccca tcatgcagaa ccagagcgtg gactgggagc cctccaccga gaagatcacc 3901 gccagcgacg gcgtgctgaa gggcgacgtg accatgtacc tgaagctgga gggcggcggc 3961 aaccacaagt gccagatgaa gaccacctac aaggccgcca aggagatcct ggagatgccc 4021 ggcgaccact acatcggcca caggctggtg aggaagaccg agggcaacat caccgagcag 4081 gtggaggacg ccgtggccca ctactcaatg ttgccttccc tgggaggata cccatcacac 4141 tggcgaccac tcgaggccag cccgccgtcc cccgcgcgat ggagcctggg acggaagcgc 4201 agagccgacg gcaggcgctg gaggcccgaa gacgccgagg aggcagagca ccgcggcgcc 4261 gagcgcagac ccgagagctt taccactcct gaaggcccta aaccccgttc cagatgctct 4321 gactgggcaa gtgcagttga agaagatgaa atgaggacca gagttaacaa agaaatggca 4381 agatataaaa ggaaactcct catcaatgac tttggaagag agagaaaatc atcatcagga 4441 agttctgatt caaaggagtc tatgtctact gtgccggctg actaagttta aacgaattcg 4501 tcgagggacc taataacttc gtatagcata cattatacga agttatacat gtttaagggt 4561 tccggttcca ctaggtacaa ttcgatatca agcttatcga taatcaacct ctggattaca 4621 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 4681 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 4741 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 4801 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 4861 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca 4921 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 4981 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga 5041 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 5101 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga 5161 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacctcga tcgagaccta 5221 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg 5281 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga 5341 ccaatgactt acaaggcagc tgacgcgtga ttttaaaaga aaagggggga ctggaagggc 5401 taattcactc ccactgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc 5461 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg 5521 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag 5581 acccttttag tcagtgtgga aaatctctag cagcatgtga gcaaaaggcc agcaaaaggc 5641 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 5701 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 5761 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 5821 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 5881 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 5941 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 6001 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 6061 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 6121 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 6181 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 6241 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 6301 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 6361 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 6421 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 6481 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 6541 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 6601 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 6661 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 6721 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 6781 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 6841 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 6901 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 6961 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 7021 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 7081 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 7141 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 7201 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 7261 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 7321 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 7381 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgac //