LOCUS       sequence-160371-        7426 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Fluorescent probe for S/G2 cell cycle transition.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2016 Oct 31. doi: 10.1038/nmeth.4045."
COMMENT     Sequence Label: mKO2-SLBP(18-126)
FEATURES             Location/Qualifiers
     source          1..7426
                     /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
                     /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    complement(2759..2792)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     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"
     CDS             3451..4101
                     /codon_start=1
                     /product="humanized monomeric Kusabira-Orange fluorescent 
                     protein"
                     /label=hmKO
                     /note="mammalian codon-optimized"
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQMKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAH"
     protein_bind    complement(4513..4546)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     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(join(6433..7224,7225..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
//
