LOCUS       EW369_CMV-TO-ZNF35(5-8) 6331 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
     promoter        1..628
                     /label="CMV"
     enhancer        1..727
                     /label="CMV promoter (with TetOs)"
     primer          80..102
                     /label="EK_CMVFa"
                     /note="sequence: cgttacataacttacggtaaatg"
     primer          156..175
                     /label="EK_CMVFb"
                     /note="sequence: tatgttcccatagtaacgcc"
     primer          232..252
                     /label="EK_CMVFc"
                     /note="sequence: tggcagtacatcaagtgtatc"
     primer          308..326
                     /label="EK_CMVFd"
                     /note="sequence: ttatgcccagtacatgacc"
     primer          384..401
                     /label="EK_CMVFe"
                     /note="sequence: gtgatgcggttttggcag"
     misc_feature    486..486
                     /label="A"
     primer          538..558
                     /label="CMV-Forward Genewiz"
                     /note="sequence: CGCAAATGGGCGGTAGGCGTG"
     TATA_signal     573..579
                     /label="TATA box"
     misc_feature    589..607
                     /label="TetO"
     misc_feature    589..628
                     /label="TetR binding sites"
     misc_feature    610..628
                     /label="TetO"
     primer          689..709
                     /label="CMVF1"
                     /note="sequence: tgacctccatagaagacaccg"
     primer          710..725
                     /label="ToFlpF"
                     /note="sequence: GGACCGATCCAGCCTC"
     primer          complement(732..762)
                     /label="EKo272.2"
                     /note="sequence: catggtggcgAAGCTTAAGTTTAAACGCTAG"
     primer          complement(737..751)
                     /label="EKo283.2"
                     /note="sequence: agcttaagtttaaac"
     misc_feature    752..759
                     /label="Kozak"
     misc_feature    760..762
                     /label="start codon"
     primer          760..780
                     /label="EWo50.1"
                     /note="sequence: atgTACGAGTGTAATGAGTGC"
     CDS             760..1419
                     /translation="MYECNECGKTFTRSSNLIVHQRIHTGEKPFACNDCGKAFTQSANLIVHQRSHTGEKPYECKECGKAFSCFSHLIVHQRIHTAEKPYDCSECGKAFSQLSCLIVHQRIHTGERPYMCVECGKCFGRSSHLLQHQRIHTGEKPYVCSVCGKAFSQSSVLSKHRRIHTGEKPYECNECGKAFRVSSDLAQHHKIHTGEKPHECLECRKAFTQLSHLIQHQRIH"
     misc_feature    763..1083
                     /label="ZNF35 ZF Domains 5-8"
     primer          970..987
                     /label="EWo51"
                     /note="sequence: AGCCACCTCATTGTACAC"
     primer          complement(980..996)
                     /label="EWo126"
                     /note="sequence: TATGCGCTGGTGTACAA"
     misc_feature    1084..1098
                     /label="Canonical ZF Linker"
     primer          complement(1086..1100)
                     /label="EWo100.2"
                     /note="sequence: TAGGGGCGTTCGCCC"
     primer          1086..1118
                     /label="EWo100.3"
                     /note="sequence: GGGCGAACGCCCCTATATGTGCGTAGAGTGCGG"
     misc_feature    1099..1419
                     /label="ZNF250 ZF Domains 1-4"
     primer          complement(1414..1428)
                     /label="EWo196.1"
                     /note="sequence: tgcagcagcGTGAAT"
     primer          1414..1469
                     /label="EWo196.2"
                     /note="sequence: ATTCACgctgctgcaTGGGAGCTAggaATGtctggcggatctGAGGGCCAGAGCGA"
     CDS             1420..1455
                     /label="Translation 1420-1455"
                     /translation="AAAWELGMSGGS"
     misc_feature    1456..1603
                     /label="N (NCOA3) Activation Domain"
     misc_feature    1456..1911
                     /label="NZF Domain"
     CDS             1456..1911
                     /label="Translation 1456-1911"
                     /translation="EGQSDERALLDQLHTLLSNTDATGLEEIDRALGIPELVNQGQALEPKQDGGWGSMAEEFVTLKDVGMDFTLGDWEQLGLEQGDTFWDTALDNCQDLFLLDPPGGGDGLGLGHEKFPSDLDLDMFNGSLECDMESIIRSELMDADGLDFNFDS"
     primer          complement(1559..1573)
                     /label="EW_NZF_R"
                     /note="sequence: GGTTCACCAGCTCGG"
     misc_feature    1618..1761
                     /label="Z (ZNF473) Activation Domain"
     misc_feature    1786..1911
                     /label="F (FOXO3) Activation Domain"
     CDS             1912..1914
                     /label="Translation 1912-1914"
                     /translation="*"
     misc_feature    1920..1938
                     /label="EWf10"
     primer          complement(1928..1949)
                     /label="pNK018"
                     /note="sequence: gagatgagtttttgttcGGGCC"
     CDS             1933..1965
                     /label="c-myc epitope"
     misc_feature    1965..1967
                     /label="New Stop"
     CDS             1978..1995
                     /label="Polyhistidine tag"
     misc_feature    1996..1998
                     /label="cya"
     primer          complement(1999..2017)
                     /label="ToFlpR"
                     /note="sequence: CTGATCAGCGGGTTTAAAC"
     polyA_signal    2017..2231
                     /label="BGH polyadenylation sequence"
     primer          complement(2018..2035)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
     primer          complement(2097..2116)
                     /label="BghR1"
                     /note="sequence: attaggaaaggacagtggga"
     primer          2208..2226
                     /label="bghF"
                     /note="sequence: AGACAATAGCAGGCATGCT"
     polyA_signal    2264..2287
                     /label="BGH pA"
     primer          2288..2306
                     /label="EKo293.3"
                     /note="sequence: cttctgaggcggaaagaac"
     primer          complement(2366..2381)
                     /label="FlpInR"
                     /note="sequence: cgcgtaaccaccacac"
     primer          2499..2517
                     /label="FrtF"
                     /note="sequence: agggttccgatttagtgct"
     misc_feature    2571..2618
                     /label="FRT"
     gene            2626..3646
                     /label="Hygromycin"
     primer          3574..3592
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
     polyA_signal    3778..3908
                     /label="SV40 pA"
     primer          complement(3791..3816)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
     primer          complement(3815..3834)
                     /label="SV40pA-R Genewiz"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
     primer          complement(3948..3964)
                     /label="M13R Genewiz"
                     /note="sequence: CAGGAAACAGCTATGAC"
     primer          complement(3962..3981)
                     /label="M13-48REV Genewiz"
                     /note="sequence: CGGATAACAATTTCACACAG"
     primer          4240..4259
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
     rep_origin      4291..4964
                     /label="pUC ori"
     primer          complement(4411..4430)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
     CDS             complement(5109..5969)
                     /label="Amp (bla)"
     promoter        complement(5970..6068)
                     /label="bla promoter"
     primer          6181..6201
                     /label="FlpInF1"
                     /note="sequence: agtatctgctccctgcttgtg"
     primer          6245..6262
                     /label="FlpInF"
                     /note="sequence: aacaaggcaaggcttgac"
     primer          6316..1
                     /label="EKo279.1"
                     /note="sequence: cgggccagatatacgcg"
     primer          6317..32
                     /label="EKo272.1"
                     /note="sequence: gggccagatatacgcgttGACATTGATTATTGACTAGTTATTAATAG"
ORIGIN
        1 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc cctatcagtg
      601 atagagatct ccctatcagt gatagagatc gtcgacgagc tcgtttagtg aaccgtcaga
      661 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca
      721 gcctccggac tctagcgttt aaacttaagc ttcgccacca tgTACGAGTG TAATGAGTGC
      781 GGTAAGACCT TCACACGATC TTCTAACCTT ATTGTGCATC AACGAATCCA TACAGGCGAG
      841 AAGCCCTTTG CCTGCAATGA CTGTGGGAAG GCTTTTACTC AGAGTGCCAA CTTGATCGTG
      901 CATCAGAGAA GCCATACGGG TGAGAAGCCC TATGAATGCA AGGAATGCGG AAAGGCCTTC
      961 TCCTGCTTCA GCCACCTCAT TGTACACCAG CGCATACACA CGGCTGAGAA GCCATACGAT
     1021 TGCTCTGAAT GTGGAAAGGC CTTTAGTCAG CTGTCTTGCT TGATTGTGCA TCAGAGAATT
     1081 CACACGGGCG AACGCCCCTA TATGTGCGTA GAGTGCGGTA AATGTTTTGG GCGGAGTTCA
     1141 CATCTTTTGC AGCACCAACG TATTCACACA GGCGAGAAAC CCTATGTTTG CAGCGTGTGC
     1201 GGTAAGGCGT TTAGCCAGTC CTCAGTCCTT TCCAAACACC GCCGCATACA TACTGGTGAG
     1261 AAGCCATATG AATGTAATGA ATGTGGCAAG GCTTTTAGAG TGAGTTCAGA TCTCGCTCAA
     1321 CATCATAAGA TCCATACAGG AGAGAAGCCA CACGAATGTT TGGAGTGTCG AAAGGCTTTT
     1381 ACTCAGCTCA GTCACCTCAT TCAGCATCAG AGAATTCACg ctgctgcaTG GGAGCTAgga
     1441 ATGtctggcg gatctGAGGG CCAGAGCGAC GAGAGAGCCC TGCTGGACCA GCTGCACACC
     1501 CTGCTGAGCA ACACCGACGC CACCGGCCTG GAGGAGATCG ACAGAGCCCT GGGCATCCCC
     1561 GAGCTGGTGA ACCAGGGCCA GGCCCTGGAG CCCAAGCAGG ACGGAGGCTG GGGATCTATG
     1621 GCCGAGGAAT TCGTGACCCT GAAGGACGTG GGCATGGACT TCACCCTGGG CGACTGGGAG
     1681 CAGCTGGGCC TGGAGCAGGG CGACACCTTC TGGGACACCG CCCTGGACAA CTGCCAGGAC
     1741 CTGTTCCTGC TGGACCCCCC CGGCGGAGGA GACGGCCTTG GTTTGGGACA CGAGAAGTTC
     1801 CCCAGCGACC TGGACCTGGA CATGTTCAAC GGCAGCCTGG AGTGCGACAT GGAGAGCATC
     1861 ATCAGAAGCG AGCTGATGGA CGCCGACGGC CTGGACTTCA ACTTCGACAG Ctaggccgct
     1921 cgaggagggc ccgaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat
     1981 catcataatc atcattgagt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2041 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2101 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2161 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2221 catgctgggg atgcggtggg ctctatggag atctatcgat gttctgggga tgcggtgggc
     2281 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2341 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2401 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2461 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2521 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtaccta gaagttccta
     2581 ttccgaagtt cctattctct agaaagtata ggaacttcct tggccaaaaa gcctgaactc
     2641 accgcgacgt ctgtcgagaa gtttctgatc gaaaagttcg acagcgtctc cgacctgatg
     2701 cagctctcgg agggcgaaga atctcgtgct ttcagcttcg atgtaggagg gcgtggatat
     2761 gtcctgcggg taaatagctg cgccgatggt ttctacaaag atcgttatgt ttatcggcac
     2821 tttgcatcgg ccgcgctccc gattccggaa gtgcttgaca ttggggaatt cagcgagagc
     2881 ctgacctatt gcatctcccg ccgtgcacag ggtgtcacgt tgcaagacct gcctgaaacc
     2941 gaactgcccg ctgttctgca gccggtcgcg gaggccatgg atgcgatcgc tgcggccgat
     3001 cttagccaga cgagcgggtt cggcccattc ggaccgcaag gaatcggtca atacactaca
     3061 tggcgtgatt tcatatgcgc gattgctgat ccccatgtgt atcactggca aactgtgatg
     3121 gacgacaccg tcagtgcgtc cgtcgcgcag gctctcgatg agctgatgct ttgggccgag
     3181 gactgccccg aagtccggca cctcgtgcac gcggatttcg gctccaacaa tgtcctgacg
     3241 gacaatggcc gcataacagc ggtcattgac tggagcgagg cgatgttcgg ggattcccaa
     3301 tacgaggtcg ccaacatctt cttctggagg ccgtggttgg cttgtatgga gcagcagacg
     3361 cgctacttcg agcggaggca tccggagctt gcaggatcgc cgcggctccg ggcgtatatg
     3421 ctccgcattg gtcttgacca actctatcag agcttggttg acggcaattt cgatgatgca
     3481 gcttgggcgc agggtcgatg cgacgcaatc gtccgatccg gagccgggac tgtcgggcgt
     3541 acacaaatcg cccgcagaag cgcggccgtc tggaccgatg gctgtgtaga agtactcgcc
     3601 gatagtggaa accgacgccc cagcactcgt ccgagggcaa aggaatagca cgtactacga
     3661 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     3721 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     3781 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3841 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     3901 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     3961 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4021 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4081 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4141 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4201 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4261 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4321 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4381 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4441 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4501 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4561 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4621 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4681 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4741 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4801 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4861 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4921 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4981 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5041 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5101 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5161 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5221 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5281 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5341 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5401 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5461 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5521 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5581 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5641 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5701 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5761 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5821 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5881 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5941 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6001 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6061 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     6121 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     6181 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     6241 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     6301 ttgcgctgct tcgtacgggc cagatatacg c
//