LOCUS       EW499_CMV-TO-NZF-(GGS)x 5977 bp ds-DNA     circular     25-FEB-2025
DEFINITION  .
COMMENT     LOCUS       CMV-TRE+H2B+Ceru        6284 bp    DNA     circular UNA
            02-AUG-2012 DEFINITION  Concatenation of 4 sequences. ACCESSION
            VERSION KEYWORDS    . SOURCE      .   ORGANISM               .
FEATURES             Location/Qualifiers
     primer          63..82
                     /label="EK_CMVFb"
                     /note="sequence: tatgttcccatagtaacgcc"
     primer          139..159
                     /label="EK_CMVFc"
                     /note="sequence: tggcagtacatcaagtgtatc"
     primer          215..233
                     /label="EK_CMVFd"
                     /note="sequence: ttatgcccagtacatgacc"
     primer          291..308
                     /label="EK_CMVFe"
                     /note="sequence: gtgatgcggttttggcag"
     misc_feature    393..393
                     /label="A"
     primer          445..465
                     /label="CMV-Forward Genewiz"
                     /note="sequence: CGCAAATGGGCGGTAGGCGTG"
     TATA_signal     480..486
                     /label="TATA box"
     misc_feature    496..514
                     /label="TetO"
     misc_feature    496..535
                     /label="TetR binding sites"
     misc_feature    517..535
                     /label="TetO"
     primer          596..616
                     /label="CMVF1"
                     /note="sequence: tgacctccatagaagacaccg"
     primer          617..632
                     /label="ToFlpF"
                     /note="sequence: GGACCGATCCAGCCTC"
     primer          639..666
                     /label="EKbF"
                     /note="sequence: ctagcgtttaaacttaagcttcgccacc"
     primer          639..683
                     /label="EWo346.1"
                     /note="sequence: ctagcgtttaaacttaagcttcgccaccatgGAGGGCCAGAGCGA"
     primer          complement(644..658)
                     /label="EKo283.2"
                     /note="sequence: agcttaagtttaaac"
     misc_feature    659..666
                     /label="Kozak"
     misc_feature    667..669
                     /label="start codon"
     CDS             667..1125
                     /translation="MEGQSDERALLDQLHTLLSNTDATGLEEIDRALGIPELVNQGQALEPKQDGGWGSMAEEFVTLKDVGMDFTLGDWEQLGLEQGDTFWDTALDNCQDLFLLDPPGGGDGLGLGHEKFPSDLDLDMFNGSLECDMESIIRSELMDADGLDFNFDS"
     misc_feature    670..817
                     /label="N (NCOA3) Activation Domain"
     misc_feature    670..1125
                     /label="NZF Domain"
     primer          complement(773..787)
                     /label="EW_NZF_R"
                     /note="sequence: GGTTCACCAGCTCGG"
     misc_feature    832..975
                     /label="Z (ZNF473) Activation Domain"
     misc_feature    1000..1125
                     /label="F (FOXO3) Activation Domain"
     primer          complement(1108..1177)
                     /label="EWo488.1"
                     /note="sequence: tgtcgccagaattgccgctcatgccgaggcccagtgttccgccatctccccaGCTGTCGAAGTTGAAGTC"
     CDS             1126..1197
                     /label="Translation 1126-1197"
                     /translation="WGDGGTLGLGMSGNSGDSVGSWGS"
     primer          1163..1215
                     /label="EWo488.2"
                     /note="sequence: gcaattctggcgacagcgtaggctcttggggatctgagatgtggcatgaaggc"
     misc_feature    1198..1464
                     /label="FRB"
     CDS             1198..1464
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETSFNQAYGRDLMEAQEWCRKYMKSGWVKDLLQAWDLYYHVFRRISK"
     primer          complement(1381..1401)
                     /label="EWo272.1"
                     /note="sequence: ccctgatttcatgtacttcct"
     primer          1387..1421
                     /label="EWo272.2"
                     /note="sequence: tacatgaaatcagggtgggtcaaggacctcctcca"
     misc_feature    1417..1419
                     /label="Rapalog Mut"
     primer          complement(1446..1491)
                     /label="EWo488.3"
                     /note="sequence: ttgttcgggccctcctcgagcggcctactttgagattcgtcggaac"
     CDS             1465..1467
                     /label="Translation 1465-1467"
                     /translation="*"
     primer          complement(1470..1491)
                     /label="EKbR"
                     /note="sequence: ttgttcGGGCCctcctcgagcg"
     primer          complement(1481..1502)
                     /label="pNK018"
                     /note="sequence: gagatgagtttttgttcGGGCC"
     CDS             1486..1518
                     /label="c-myc epitope"
     misc_feature    1518..1520
                     /label="New Stop"
     CDS             1531..1548
                     /label="Polyhistidine tag"
     misc_feature    1549..1551
                     /label="cya"
     primer          complement(1552..1570)
                     /label="ToFlpR"
                     /note="sequence: CTGATCAGCGGGTTTAAAC"
     polyA_signal    1570..1784
                     /label="BGH polyadenylation sequence"
     primer          complement(1571..1588)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
     primer          complement(1650..1669)
                     /label="BghR1"
                     /note="sequence: attaggaaaggacagtggga"
     primer          1761..1779
                     /label="bghF"
                     /note="sequence: AGACAATAGCAGGCATGCT"
     polyA_signal    1817..1840
                     /label="BGH pA"
     primer          1841..1859
                     /label="EKo293.3"
                     /note="sequence: cttctgaggcggaaagaac"
     primer          complement(1919..1934)
                     /label="FlpInR"
                     /note="sequence: cgcgtaaccaccacac"
     primer          2052..2070
                     /label="FrtF"
                     /note="sequence: agggttccgatttagtgct"
     misc_feature    2124..2171
                     /label="FRT"
     gene            2179..3199
                     /label="Hygromycin"
     primer          3127..3145
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
     polyA_signal    3331..3461
                     /label="SV40 pA"
     primer          complement(3344..3369)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
     primer          complement(3368..3387)
                     /label="SV40pA-R Genewiz"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
     primer          complement(3501..3517)
                     /label="M13R Genewiz"
                     /note="sequence: CAGGAAACAGCTATGAC"
     primer          complement(3515..3534)
                     /label="M13-48REV Genewiz"
                     /note="sequence: CGGATAACAATTTCACACAG"
     primer          3793..3812
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
     rep_origin      3844..4517
                     /label="pUC ori"
     primer          complement(3964..3983)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
     CDS             complement(4662..5522)
                     /label="Amp (bla)"
     promoter        complement(5523..5621)
                     /label="bla promoter"
     primer          5734..5754
                     /label="FlpInF1"
                     /note="sequence: agtatctgctccctgcttgtg"
     primer          5798..5815
                     /label="FlpInF"
                     /note="sequence: aacaaggcaaggcttgac"
     primer          5869..5885
                     /label="EKo279.1"
                     /note="sequence: cgggccagatatacgcg"
     primer          5870..5916
                     /label="EKo272.1"
                     /note="sequence: gggccagatatacgcgttGACATTGATTATTGACTAGTTATTAATAG"
     promoter        5885..535
                     /label="CMV"
     enhancer        5885..634
                     /label="CMV promoter (with TetOs)"
     primer          5964..9
                     /label="EK_CMVFa"
                     /note="sequence: cgttacataacttacggtaaatg"
ORIGIN
        1 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
       61 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      121 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      181 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      241 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      301 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      361 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      421 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      481 atataagcag agctctccct atcagtgata gagatctccc tatcagtgat agagatcgtc
      541 gacgagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
      601 tccatagaag acaccgggac cgatccagcc tccggactct agcgtttaaa cttaagcttc
      661 gccaccatgG AGGGCCAGAG CGACGAGAGA GCCCTGCTGG ACCAGCTGCA CACCCTGCTG
      721 AGCAACACCG ACGCCACCGG CCTGGAGGAG ATCGACAGAG CCCTGGGCAT CCCCGAGCTG
      781 GTGAACCAGG GCCAGGCCCT GGAGCCCAAG CAGGACGGAG GCTGGGGATC TATGGCCGAG
      841 GAATTCGTGA CCCTGAAGGA CGTGGGCATG GACTTCACCC TGGGCGACTG GGAGCAGCTG
      901 GGCCTGGAGC AGGGCGACAC CTTCTGGGAC ACCGCCCTGG ACAACTGCCA GGACCTGTTC
      961 CTGCTGGACC CCCCCGGCGG AGGAGACGGC CTTGGTTTGG GACACGAGAA GTTCCCCAGC
     1021 GACCTGGACC TGGACATGTT CAACGGCAGC CTGGAGTGCG ACATGGAGAG CATCATCAGA
     1081 AGCGAGCTGA TGGACGCCGA CGGCCTGGAC TTCAACTTCG ACAGCtgggg agatggcgga
     1141 acactgggcc tcggcatgag cggcaattct ggcgacagcg taggctcttg gggatctgag
     1201 atgtggcatg aaggcctgga agaggcatct cgtttgtact ttggggaaag gaacgtgaaa
     1261 ggcatgtttg aggtgctgga gcccttgcat gctatgatgg aacggGGACC Ccagactctg
     1321 aaggaaacat cctttaatca ggcctatggt cgagatttaa tggaggccca agagtggtgc
     1381 aggaagtaca tgaaatcagg gtgggtcaag gacctcctcc aagcctggga cctctattat
     1441 catgtgttcc gacgaatctc aaagtaggcc gctcgaggag ggcccgaaca aaaactcatc
     1501 tcagaagagg atctgaatag cgccgtcgac catcatcata atcatcattg agtttaaacc
     1561 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1621 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1681 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1741 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     1801 gagatctatc gatgttctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
     1861 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
     1921 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     1981 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2041 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2101 tagggtgatg gttcacgtac ctagaagttc ctattccgaa gttcctattc tctagaaagt
     2161 ataggaactt ccttggccaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg
     2221 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt
     2281 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat
     2341 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg
     2401 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca
     2461 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc
     2521 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca
     2581 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct
     2641 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg
     2701 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg
     2761 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt
     2821 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg
     2881 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag
     2941 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat
     3001 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca
     3061 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc
     3121 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact
     3181 cgtccgaggg caaaggaata gcacgtacta cgagatttcg attccaccgc cgccttctat
     3241 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3301 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     3361 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     3421 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     3481 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     3541 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     3601 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     3661 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     3721 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3781 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3841 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3901 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3961 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4021 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4081 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4141 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4201 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4261 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4321 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4381 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4441 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4501 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4561 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4621 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4681 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4741 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4801 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4861 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4921 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4981 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5041 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5101 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5161 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5221 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5281 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5341 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5401 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5461 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5521 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5581 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5641 aaagtgccac ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca
     5701 gtacaatctg ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg
     5761 aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca
     5821 attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgtacg ggccagatat
     5881 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     5941 catagcccat atatggagtt ccgcgttaca taactta
//