LOCUS       EW269_CMV-TO-Gal4-NZF[G 6112 bp ds-DNA     circular     26-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          complement(639..669)
                     /label="EKo272.2"
                     /note="sequence: catggtggcgAAGCTTAAGTTTAAACGCTAG"
     primer          641..687
                     /label="EKo37.1"
                     /note="sequence: agcgtttaaacttaagcttcgccaccatgaagctgctgagcagcatc"
     primer          complement(644..658)
                     /label="EKo283.2"
                     /note="sequence: agcttaagtttaaac"
     primer          644..665
                     /label="EWo100.1"
                     /note="sequence: gtttaaacttaagcttcgccac"
     primer          654..669
                     /label="EKo271.3"
                     /note="sequence: aagcttcgccaccatg"
     misc_feature    659..666
                     /label="Kozak"
     primer          667..691
                     /label="EKoGal4f"
                     /note="sequence: atgaagctgctgagcagcatcgagc"
     CDS             667..1107
                     /label="GAL4DBD"
     CDS             667..1143
                     /label="Translation 667-1143"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTKRSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNKDAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVSAAAGGSGGSGGS"
     primer          complement(676..691)
                     /label="EWo23.2"
                     /note="sequence: gctcgatgctgctcag"
     primer          complement(756..775)
                     /label="pPYK027"
                     /note="sequence: cccagttgttcttcaggcac"
     primer          1128..1142
                     /label="EW112.2"
                     /note="sequence: aggctctggcggatc"
     primer          complement(1128..1142)
                     /label="EW112.1"
                     /note="sequence: gatccgccagagcct"
     primer          1128..1157
                     /label="EWo152.1"
                     /note="sequence: aggctctggcggatctGAGGGCCAGAGCGA"
     misc_feature    1128..1626
                     /label="EWf10"
     misc_feature    1144..1291
                     /label="N (NCOA3) Activation Domain"
     misc_feature    1144..1599
                     /label="NZF Domain"
     CDS             1144..1599
                     /label="Translation 1144-1599"
                     /translation="EGQSDERALLDQLHTLLSNTDATGLEEIDRALGIPELVNQGQALEPKQDGGSGSMAEEFVTLKDVGMDFTLGDWEQLGLEQGDTFWDTALDNCQDLFLLDPPGGGSGGGSGHEKFPSDLDLDMFNGSLECDMESIIRSELMDADGLDFNFDS"
     primer          1201..1213
                     /label="EWo269.1"
                     /note="sequence: ACCGACGCCACCG"
     primer          complement(1284..1302)
                     /label="EWo269.2"
                     /note="sequence: TCCAGAGCCTCCGTCCTGC"
     primer          1288..1315
                     /label="EWo269.3"
                     /note="sequence: GACGGAGGCTCTGGATCTATGGCCGAGG"
     misc_feature    1306..1449
                     /label="Z (ZNF473) Activation Domain"
     primer          complement(1447..1466)
                     /label="EWo269.4"
                     /note="sequence: CCGCCAGATCCTCCGCCGGG"
     primer          1452..1490
                     /label="EWo269.5"
                     /note="sequence: CGGAGGATCTGGCGGTGGTTCTGGACACGAGAAGTTCCC"
     misc_feature    1474..1599
                     /label="F (FOXO3) Activation Domain"
     CDS             1600..1602
                     /label="Translation 1600-1602"
                     /translation="*"
     primer          complement(1605..1626)
                     /label="EKbR"
                     /note="sequence: ttgttcGGGCCctcctcgagcg"
     primer          complement(1616..1637)
                     /label="pNK018"
                     /note="sequence: gagatgagtttttgttcGGGCC"
     CDS             1621..1653
                     /label="c-myc epitope"
     misc_feature    1653..1655
                     /label="New Stop"
     CDS             1666..1683
                     /label="Polyhistidine tag"
     misc_feature    1684..1686
                     /label="cya"
     primer          complement(1687..1705)
                     /label="ToFlpR"
                     /note="sequence: CTGATCAGCGGGTTTAAAC"
     polyA_signal    1705..1919
                     /label="BGH polyadenylation sequence"
     primer          complement(1706..1723)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
     primer          complement(1785..1804)
                     /label="BghR1"
                     /note="sequence: attaggaaaggacagtggga"
     primer          1896..1914
                     /label="bghF"
                     /note="sequence: AGACAATAGCAGGCATGCT"
     polyA_signal    1952..1975
                     /label="BGH pA"
     primer          1976..1994
                     /label="EKo293.3"
                     /note="sequence: cttctgaggcggaaagaac"
     primer          complement(2054..2069)
                     /label="FlpInR"
                     /note="sequence: cgcgtaaccaccacac"
     primer          2187..2205
                     /label="FrtF"
                     /note="sequence: agggttccgatttagtgct"
     misc_feature    2259..2306
                     /label="FRT"
     primer          complement(2280..2307)
                     /label="EKo299.3"
                     /note="sequence: ggaagttcctatactttctagagaatag"
     gene            2314..3334
                     /label="Hygromycin"
     primer          2542..2563
                     /label="EKo152.1"
                     /note="sequence: cttgacattggggaattcagcg"
     primer          3262..3280
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
     polyA_signal    3466..3596
                     /label="SV40 pA"
     primer          complement(3479..3504)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
     primer          complement(3503..3522)
                     /label="SV40pA-R Genewiz"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
     primer          complement(3636..3652)
                     /label="M13R Genewiz"
                     /note="sequence: CAGGAAACAGCTATGAC"
     primer          complement(3650..3669)
                     /label="M13-48REV Genewiz"
                     /note="sequence: CGGATAACAATTTCACACAG"
     primer          3928..3947
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
     rep_origin      3979..4652
                     /label="pUC ori"
     primer          complement(4099..4118)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
     CDS             complement(4797..5657)
                     /label="Amp (bla)"
     primer          complement(4922..4940)
                     /label="EKo152.2"
                     /note="sequence: tctcgcggtatcattgcag"
     promoter        complement(5658..5756)
                     /label="bla promoter"
     primer          5869..5889
                     /label="FlpInF1"
                     /note="sequence: agtatctgctccctgcttgtg"
     primer          5933..5950
                     /label="FlpInF"
                     /note="sequence: aacaaggcaaggcttgac"
     primer_bind     5944..5963
                     /label="primer3"
     primer          5944..5963
                     /label="FlpInF2"
                     /note="sequence: gcttgaccgacaattgcatg"
     primer          complement(5948..5970)
                     /label="EKo152.4"
                     /note="sequence: gattcttcatgcaattgtcggtc"
     primer          6004..6020
                     /label="EKo279.1"
                     /note="sequence: cgggccagatatacgcg"
     primer          6005..6051
                     /label="EKo272.1"
                     /note="sequence: gggccagatatacgcgttGACATTGATTATTGACTAGTTATTAATAG"
     promoter        6020..535
                     /label="CMV"
     enhancer        6020..634
                     /label="CMV promoter (with TetOs)"
     primer          6099..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 gccaccatga agctgctgag cagcatcgag caggcctgtg acatctgccg gctgaagaaa
      721 ctgaagtgca gcaaagaaaa gcccaagtgc gccaagtgcc tgaagaacaa ctgggagtgc
      781 cggtacagcc ccaagaccaa gagaagcccc ctgaccagag cccacctgac cgaggtggaa
      841 agccggctgg aaagactgga acagctgttt ctgctgatct tcccacgcga ggacctggac
      901 atgatcctga agatggacag cctgcaggac atcaaggccc tgctgaccgg cctgttcgtg
      961 caggacaacg tgaacaagga cgccgtgacc gacagactgg ccagcgtgga aaccgacatg
     1021 cccctgaccc tgcggcagca cagaatcagc gccaccagca gcagcgagga aagcagcaac
     1081 aagggccagc ggcagctgac agtgtctgct gctgcaggcg gaagcggagg ctctggcgga
     1141 tctGAGGGCC AGAGCGACGA GAGAGCCCTG CTGGACCAGC TGCACACCCT GCTGAGCAAC
     1201 ACCGACGCCA CCGGCCTGGA GGAGATCGAC AGAGCCCTGG GCATCCCCGA GCTGGTGAAC
     1261 CAGGGCCAGG CCCTGGAGCC CAAGCAGGAC GGAGGCTCTG GATCTATGGC CGAGGAATTC
     1321 GTGACCCTGA AGGACGTGGG CATGGACTTC ACCCTGGGCG ACTGGGAGCA GCTGGGCCTG
     1381 GAGCAGGGCG ACACCTTCTG GGACACCGCC CTGGACAACT GCCAGGACCT GTTCCTGCTG
     1441 GACCCCCCCG GCGGAGGATC TGGCGGTGGT TCTGGACACG AGAAGTTCCC CAGCGACCTG
     1501 GACCTGGACA TGTTCAACGG CAGCCTGGAG TGCGACATGG AGAGCATCAT CAGAAGCGAG
     1561 CTGATGGACG CCGACGGCCT GGACTTCAAC TTCGACAGCt aggccgctcg aggagggccc
     1621 gaacaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca tcataatcat
     1681 cattgagttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     1741 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1801 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     1861 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     1921 gcggtgggct ctatggagat ctatcgatgt tctggggatg cggtgggctc tatggcttct
     1981 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2041 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2101 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2161 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2221 cccaaaaaac ttgattaggg tgatggttca cgtacctaga agttcctatt ccgaagttcc
     2281 tattctctag aaagtatagg aacttccttg gccaaaaagc ctgaactcac cgcgacgtct
     2341 gtcgagaagt ttctgatcga aaagttcgac agcgtctccg acctgatgca gctctcggag
     2401 ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc gtggatatgt cctgcgggta
     2461 aatagctgcg ccgatggttt ctacaaagat cgttatgttt atcggcactt tgcatcggcc
     2521 gcgctcccga ttccggaagt gcttgacatt ggggaattca gcgagagcct gacctattgc
     2581 atctcccgcc gtgcacaggg tgtcacgttg caagacctgc ctgaaaccga actgcccgct
     2641 gttctgcagc cggtcgcgga ggccatggat gcgatcgctg cggccgatct tagccagacg
     2701 agcgggttcg gcccattcgg accgcaagga atcggtcaat acactacatg gcgtgatttc
     2761 atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc
     2821 agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt gggccgagga ctgccccgaa
     2881 gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg tcctgacgga caatggccgc
     2941 ataacagcgg tcattgactg gagcgaggcg atgttcgggg attcccaata cgaggtcgcc
     3001 aacatcttct tctggaggcc gtggttggct tgtatggagc agcagacgcg ctacttcgag
     3061 cggaggcatc cggagcttgc aggatcgccg cggctccggg cgtatatgct ccgcattggt
     3121 cttgaccaac tctatcagag cttggttgac ggcaatttcg atgatgcagc ttgggcgcag
     3181 ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg tcgggcgtac acaaatcgcc
     3241 cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag tactcgccga tagtggaaac
     3301 cgacgcccca gcactcgtcc gagggcaaag gaatagcacg tactacgaga tttcgattcc
     3361 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     3421 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     3481 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3541 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     3601 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     3661 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     3721 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     3781 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3841 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     3901 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     3961 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4021 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4081 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4141 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     4201 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     4261 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     4321 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4381 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4441 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4501 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4561 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4621 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4681 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4741 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4801 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4861 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4921 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4981 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5041 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5101 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     5161 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     5221 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     5281 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     5341 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5401 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5461 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5521 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5581 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5641 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5701 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5761 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc
     5821 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     5881 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     5941 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     6001 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
     6061 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact ta
//