LOCUS       V011877                 7475 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V011877
VERSION     V011877
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."
            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"
                     /label="Protein Tat"
     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
//
