LOCUS       V012513                 9033 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V012513
VERSION     V012513
KEYWORDS    pLL3.7m-Clover-Geminin(1-110)-IRES-mKO2-Cdt(30-120)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 9033)
  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 9033)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9033)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Methods. 2016 Oct 31. doi: 10.1038/nmeth.4045."
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9033
                     /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    1433..1666
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             1851..1895
                     /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             2044..2085
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2193..2310
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2369..2681
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das
                     et al., 1988)"
     primer_bind     2698..2714
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     protein_bind    2756..2789
                     /label="loxP"
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (GCATACAT)."
     enhancer        2905..3208
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3209..3412
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             3444..4160
                     /label="Clover"
                     /note="bright green-yellow fluorescent protein derived from
                     GFP (Lam et al., 2012)"
     CDS             4191..4520
                     /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"
     misc_feature    4525..5105
                     /label="IRES2"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             5112..5762
                     /label="mKO2"
                     /note="monomeric Kusabira-Orange 2 fluorescent protein"
     CDS             5817..6089
                     /label="Cdt1 (30-120)"
                     /note="degron consisting of residues 30-120 of human Cdt1
                     (Zielke and Edgar, 2015)"
     protein_bind    complement(6120..6153)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6209..6797
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(6800..6816)
                     /label="KS primer"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     LTR             7039..7219
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7281..7869)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(8043..8900)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(8901..9005)
                     /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 ccgctgatct tcagacctgg aggaggagat atgagggaca
     1321 attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac
     1381 ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt
     1441 tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga
     1501 cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg
     1561 ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg
     1621 caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt
     1681 gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat
     1741 ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt
     1801 acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac
     1861 aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt
     1921 ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag
     1981 tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc
     2041 agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg
     2101 gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg cgtgcgccaa
     2161 ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg gattgggggg
     2221 tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac taaagaatta
     2281 caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag cagagatcca
     2341 gtttggttag taccgggccc gctctagaga tccgacgcgc catctctagg cccgcgccgg
     2401 ccccctcgca cagacttgtg ggagaagctc ggctactccc ctgccccggt taatttgcat
     2461 ataatatttc ctagtaacta tagaggctta atgtgcgata aaagacagat aatctgttct
     2521 ttttaatact agctacattt tacatgatag gcttggattt ctataagaga tacaaatact
     2581 aaattattat tttaaaaaac agcacaaaag gaaactcacc ctaactgtaa agtaattgtg
     2641 tgttttgaga ctataaatat cccttggaga aaagccttgt taacgcgcgg tgaccctcga
     2701 ggtcgacggt atcgataagc tcgcttcacg agattccagc aggtcgaggg acctaataac
     2761 ttcgtatagc atacattata cgaagttata ttaagggttc caagcttaag cggccgcgtg
     2821 gataaccgta ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt
     2881 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     2941 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     3001 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     3061 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     3121 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     3181 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     3241 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     3301 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     3361 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
     3421 gaaccgtcag atccgctagc accatggtga gcaagggcga ggagctgttc accggggtgg
     3481 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtccgcggcg
     3541 agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc accaccggca
     3601 agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcgtg gcctgcttca
     3661 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     3721 acgtccagga gcgcaccatc tctttcaagg acgacggtac ctacaagacc cgcgccgagg
     3781 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     3841 aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac aacgtctata
     3901 tcacggccga caagcagaag aacggcatca aggctaactt caagatccgc cacaacgttg
     3961 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     4021 ccgtgctgct gcccgacaac cactacctga gccatcagtc cgccctgagc aaagacccca
     4081 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg attacacatg
     4141 gcatggacga gctgtacaag gatatccatg cacactggcg gccgctcgag atgaatccca
     4201 gtatgaagca gaaacaagaa gaaatcaaag agaatataaa gaatagttct gtcccaagaa
     4261 gaactctgaa gatgattcag ccttctgcat ctggatctct tgttggaaga gaaaatgagc
     4321 tgtccgcagg cttgtccaaa aggaaacatc ggaatgacca cttaacatct acaacttcca
     4381 gccctggggt tattgtccca gaatctagtg aaaataaaaa tcttggagga gtcacccagg
     4441 agtcatttga tcttatgatt aaagaaaatc catcctctca gtattggaag gaagtggcag
     4501 aaaaacggag aaaggcgctg taagcccctc tccctccccc ccccctaacg ttactggccg
     4561 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     4621 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     4681 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     4741 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     4801 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     4861 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     4921 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     4981 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     5041 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatgg
     5101 ccacagaatt catggtgagc gtgatcaagc ccgagatgaa gatgaggtac tacatggacg
     5161 gctccgtcaa tgggcatgag ttcacaatcg agggtgaggg cacaggcaga ccttacgagg
     5221 gacatcagga gatgacactg cgcgtcacaa tggccgaggg cgggccaatg cctttcgcct
     5281 tcgacctggt gtcccacgtg ttctgttacg gccacagagt ttttaccaag tacccagaag
     5341 agatcccaga ctatttcaag caggcctttc ctgagggcct gtcctgggag aggtccctgg
     5401 agttcgagga cggcggctcc gcctccgtga gcgcccacat cagcctgagg ggcaacacct
     5461 tctaccacaa gtccaagttc accggcgtga acttccccgc cgacggcccc atcatgcaga
     5521 accagagcgt ggactgggag ccctccaccg agaagatcac cgccagcgac ggcgtgctga
     5581 agggcgacgt gaccatgtac ctgaagctgg agggcggcgg caaccacaag tgccagatga
     5641 agaccaccta caaggccgcc aaggagatcc tggagatgcc cggcgaccac tacatcggcc
     5701 acaggctggt gaggaagacc gagggcaaca tcaccgagca ggtggaggac gccgtggccc
     5761 actactcaat gttgccttcc ctgggcgata tccatgcaca ctggcggccg ctcgagccca
     5821 gccccgccag gcccgcactc cgcgccccgg cctccgctac cagtggcagc cgcaagcgcg
     5881 cccgcccgcc cgccgccccc ggacgcgacc aggccaggcc accggcccgc aggagactgc
     5941 ggctgtcggt ggacgaggtt tccagcccca gtacccccga ggccccagac atcccagcct
     6001 gcccttctcc gggccagaag ataaagaaat ccaccccggc agcaggtcag ccgccccacc
     6061 tgacatccgc gcaggaccag gacaccatct gagtttaaac gaattcgtcg agggacctaa
     6121 taacttcgta tagcatacat tatacgaagt tatacatgtt taagggttcc ggttccacta
     6181 ggtacaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     6241 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     6301 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     6361 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     6421 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     6481 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     6541 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     6601 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     6661 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     6721 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     6781 ttgggccgcc tccccgcatc gataccgtcg acctcgatcg agacctagaa aaacatggag
     6841 caatcacaag tagcaataca gcagctacca atgctgattg tgcctggcta gaagcacaag
     6901 aggaggagga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca atgacttaca
     6961 aggcagctga cgcgtgattt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     7021 ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
     7081 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
     7141 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
     7201 gtgtggaaaa tctctagcag catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     7261 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     7321 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     7381 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     7441 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     7501 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     7561 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     7621 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     7681 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     7741 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     7801 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     7861 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     7921 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     7981 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     8041 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     8101 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     8161 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     8221 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     8281 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     8341 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     8401 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     8461 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     8521 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     8581 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     8641 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     8701 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     8761 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     8821 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     8881 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     8941 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     9001 ccgcgcacat ttccccgaaa agtgccacct gac
//
