LOCUS       EW368_CMV-TO-UtroUpZF-d 6211 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          complement(644..658)
                     /label="EKo283.2"
                     /note="sequence: agcttaagtttaaac"
     misc_feature    659..666
                     /label="Kozak"
     misc_feature    667..669
                     /label="start codon"
     CDS             667..669
                     /label="Translation 667-669"
                     /translation="M"
     misc_feature    670..1206
                     /label="UtroUp"
     CDS             670..1206
                     /translation="ILDRPYACPVESCDRRFSRSDNLVRHIRIHTGQKPFQCRICMRNFSRSDHLTTHNRTHTGEKPFACDICGRKFADPGHLVRHNRIHTGEKPFACPVESCDRRFSRSDELTRHIRIHTGQKPFQCRICMRNFSSRDVLRRHNRTHTGEKPFACDICGRKFASRDVLRRHNRIHLRQNDLE"
     CDS             1207..1242
                     /label="Translation 1207-1242"
                     /translation="AAAWELGMSGGS"
     primer          1243..1256
                     /label="EWo211"
                     /note="sequence: GAGGGCCAGAGCGA"
     misc_feature    1243..1390
                     /label="N (NCOA3) Activation Domain"
     misc_feature    1243..1698
                     /label="NZF Domain"
     CDS             1243..1698
                     /label="Translation 1243-1698"
                     /translation="EGQSDERALLDQLHTLLSNTDATGLEEIDRALGIPELVNQGQALEPKQDGGWGSMAEEFVTLKDVGMDFTLGDWEQLGLEQGDTFWDTALDNCQDLFLLDPPGGGDGLGLGHEKFPSDLDLDMFNGSLECDMESIIRSELMDADGLDFNFDS"
     primer          complement(1346..1360)
                     /label="EW_NZF_R"
                     /note="sequence: GGTTCACCAGCTCGG"
     misc_feature    1405..1548
                     /label="Z (ZNF473) Activation Domain"
     misc_feature    1573..1698
                     /label="F (FOXO3) Activation Domain"
     CDS             1699..1701
                     /label="Translation 1699-1701"
                     /translation="*"
     primer          complement(1715..1736)
                     /label="pNK018"
                     /note="sequence: gagatgagtttttgttcGGGCC"
     CDS             1720..1752
                     /label="c-myc epitope"
     misc_feature    1752..1754
                     /label="New Stop"
     CDS             1765..1782
                     /label="Polyhistidine tag"
     misc_feature    1783..1785
                     /label="cya"
     primer          complement(1786..1804)
                     /label="ToFlpR"
                     /note="sequence: CTGATCAGCGGGTTTAAAC"
     polyA_signal    1804..2018
                     /label="BGH polyadenylation sequence"
     primer          complement(1805..1822)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
     primer          complement(1884..1903)
                     /label="BghR1"
                     /note="sequence: attaggaaaggacagtggga"
     primer          1995..2013
                     /label="bghF"
                     /note="sequence: AGACAATAGCAGGCATGCT"
     polyA_signal    2051..2074
                     /label="BGH pA"
     primer          2075..2093
                     /label="EKo293.3"
                     /note="sequence: cttctgaggcggaaagaac"
     primer          complement(2153..2168)
                     /label="FlpInR"
                     /note="sequence: cgcgtaaccaccacac"
     primer          2286..2304
                     /label="FrtF"
                     /note="sequence: agggttccgatttagtgct"
     misc_feature    2358..2405
                     /label="FRT"
     gene            2413..3433
                     /label="Hygromycin"
     primer          3361..3379
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
     polyA_signal    3565..3695
                     /label="SV40 pA"
     primer          complement(3578..3603)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
     primer          complement(3602..3621)
                     /label="SV40pA-R Genewiz"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
     primer          complement(3735..3751)
                     /label="M13R Genewiz"
                     /note="sequence: CAGGAAACAGCTATGAC"
     primer          complement(3749..3768)
                     /label="M13-48REV Genewiz"
                     /note="sequence: CGGATAACAATTTCACACAG"
     primer          4027..4046
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
     rep_origin      4078..4751
                     /label="pUC ori"
     primer          complement(4198..4217)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
     CDS             complement(4896..5756)
                     /label="Amp (bla)"
     promoter        complement(5757..5855)
                     /label="bla promoter"
     primer          5968..5988
                     /label="FlpInF1"
                     /note="sequence: agtatctgctccctgcttgtg"
     primer          6032..6049
                     /label="FlpInF"
                     /note="sequence: aacaaggcaaggcttgac"
     primer          6103..6119
                     /label="EKo279.1"
                     /note="sequence: cgggccagatatacgcg"
     primer          6104..6150
                     /label="EKo272.1"
                     /note="sequence: gggccagatatacgcgttGACATTGATTATTGACTAGTTATTAATAG"
     promoter        6119..535
                     /label="CMV"
     enhancer        6119..634
                     /label="CMV promoter (with TetOs)"
     primer          6198..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 TATTAGACCG GCCCTATGCG TGCCCAGTCG AATCTTGCGA TCGTCGCTTC
      721 AGCAGAAGTG ATAACTTAGT CAGACACATA AGGATCCACA CGGGTCAAAA ACCGTTCCAA
      781 TGCAGGATTT GTATGAGGAA TTTCTCGCGG AGTGACCACC TCACGACACA CAACCGCACT
      841 CACACCGGAG AGAAACCATT TGCATGCGAT ATATGCGGCA GGAAGTTCGC TGATCCGGGA
      901 CATCTAGTCC GTCACAATCG GATACACACC GGGGAAAAGC CGTTTGCGTG CCCTGTGGAA
      961 AGCTGCGATC GAAGATTCTC GCGCTCCGAC GAACTAACTC GCCATATTAG AATACATACC
     1021 GGACAGAAGC CCTTTCAGTG TCGGATCTGC ATGCGTAACT TTTCGTCAAG GGACGTACTG
     1081 AGAAGACATA ACAGGACACA TACCGGGGAA AAGCCATTTG CGTGCGACAT ATGTGGGAGA
     1141 AAGTTCGCTT CCCGGGATGT ACTTAGGAGG CATAATCGGA TTCATCTACG CCAAAATGAC
     1201 TTGGAAgctg ctgcaTGGGA GCTAggaATG tctggcggat ctGAGGGCCA GAGCGACGAG
     1261 AGAGCCCTGC TGGACCAGCT GCACACCCTG CTGAGCAACA CCGACGCCAC CGGCCTGGAG
     1321 GAGATCGACA GAGCCCTGGG CATCCCCGAG CTGGTGAACC AGGGCCAGGC CCTGGAGCCC
     1381 AAGCAGGACG GAGGCTGGGG ATCTATGGCC GAGGAATTCG TGACCCTGAA GGACGTGGGC
     1441 ATGGACTTCA CCCTGGGCGA CTGGGAGCAG CTGGGCCTGG AGCAGGGCGA CACCTTCTGG
     1501 GACACCGCCC TGGACAACTG CCAGGACCTG TTCCTGCTGG ACCCCCCCGG CGGAGGAGAC
     1561 GGCCTTGGTT TGGGACACGA GAAGTTCCCC AGCGACCTGG ACCTGGACAT GTTCAACGGC
     1621 AGCCTGGAGT GCGACATGGA GAGCATCATC AGAAGCGAGC TGATGGACGC CGACGGCCTG
     1681 GACTTCAACT TCGACAGCta ggccgctcga ggagggcccg aacaaaaact catctcagaa
     1741 gaggatctga atagcgccgt cgaccatcat cataatcatc attgagttta aacccgctga
     1801 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1861 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1921 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1981 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggagatc
     2041 tatcgatgtt ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2101 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2161 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2221 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2281 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2341 gatggttcac gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga
     2401 acttccttgg ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     2461 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     2521 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     2581 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     2641 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     2701 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     2761 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     2821 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     2881 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     2941 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     3001 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     3061 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     3121 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     3181 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     3241 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     3301 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     3361 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     3421 agggcaaagg aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3481 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3541 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     3601 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3661 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     3721 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3781 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3841 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3901 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3961 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4021 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4081 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4141 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4201 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4261 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4321 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4381 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4441 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4501 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4561 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4621 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4681 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4741 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4801 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4861 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4921 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4981 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5041 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5101 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5161 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5221 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5281 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5341 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5401 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5461 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5521 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5581 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5641 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5701 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5761 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5821 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5881 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     5941 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     6001 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     6061 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg tacgggccag atatacgcgt
     6121 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     6181 ccatatatgg agttccgcgt tacataactt a
//