LOCUS       EW415_PGK-ZNF35(5-8)-IK 5927 bp ds-DNA     circular     25-FEB-2025
DEFINITION  .
COMMENT     Concatenation of 4 sequences.
FEATURES             Location/Qualifiers
     CDS             complement(171..1031)
                     /label="Amp (bla)"
     promoter        complement(1032..1130)
                     /label="bla promoter"
     primer          1243..1263
                     /label="FlpInF1"
                     /note="sequence: agtatctgctccctgcttgtg"
     primer          1307..1324
                     /label="FlpInF"
                     /note="sequence: aacaaggcaaggcttgac"
     misc_feature    1396..1909
                     /label="PGK promoter"
     misc_feature    1515..1515
                     /label="change to A?"
     primer          1834..1850
                     /label="PGK2"
                     /note="sequence: CATTCTGCACGCTTCAA"
     primer          1903..1928
                     /label="EWo209"
                     /note="sequence: cctgcagcccaagcttcgccaccatg"
     primer          1914..1993
                     /label="EWo153.3"
                     /note="sequence: agcttcgccaccatgTACGAGTGTAATGAGTGCGGTAAGACCTTCACACGATCTTCTAACCTTATTGTGCATCAACGAAT"
     misc_feature    1918..1925
                     /label="Kozak"
     primer          complement(1918..1997)
                     /label="EWo153.4"
                     /note="sequence: ATGGATTCGTTGATGCACAATAAGGTTAGAAGATCGTGTGAAGGTCTTACCGCACTCATTACACTCGTAcatggtggcga"
     misc_feature    1926..1928
                     /label="start codon"
     CDS             1926..1928
                     /label="Translation 1926-1928"
                     /translation="M"
     misc_feature    1929..2249
                     /label="ZNF35 ZF Domains 5-8"
     CDS             1929..2264
                     /label="Translation 1929-2264"
                     /translation="YECNECGKTFTRSSNLIVHQRIHTGEKPFACNDCGKAFTQSANLIVHQRSHTGEKPYECKECGKAFSCFSHLIVHQRIHTAEKPYDCSECGKAFSQLSCLIVHQRIHTGERP"
     primer          complement(1981..2012)
                     /label="EWo153.1"
                     /note="sequence: GGGCTTCTCGCCTGTATGGATTCGTTGATGCA"
     primer          2136..2153
                     /label="EWo51"
                     /note="sequence: AGCCACCTCATTGTACAC"
     primer          complement(2146..2162)
                     /label="EWo126"
                     /note="sequence: TATGCGCTGGTGTACAA"
     primer          complement(2230..2249)
                     /label="EWo169.1"
                     /note="sequence: GTGAATTCTCTGATGCACAA"
     misc_feature    2250..2264
                     /label="Canonical ZF Linker"
     primer          complement(2250..2264)
                     /label="EWo120.1"
                     /note="sequence: GGGGCGTTCGCCCGT"
     misc_feature    2265..2417
                     /label="IKZF1 ZF 2-4"
     CDS             2265..2417
                     /label="Translation 2265-2417"
                     /translation="FQCNQCGASFTQKGNLLRHIKLHSGEKPFKCHLCNYACRRRDALTGHLRTH"
     primer          complement(2401..2417)
                     /label="EWo366.1"
                     /note="sequence: ATGGGTTCTGAGGTGGC"
     primer          complement(2401..2459)
                     /label="EWo415"
                     /note="sequence: GTCGCTCTGGCCCTCgctcatgccATCgcctccgctCCAgccATGGGTTCTGAGGTGGC"
     CDS             2418..2444
                     /label="Translation 2418-2444"
                     /translation="GWSGGDGMS"
     primer          2445..2458
                     /label="EWo211"
                     /note="sequence: GAGGGCCAGAGCGA"
     misc_feature    2445..2592
                     /label="N (NCOA3) Activation Domain"
     misc_feature    2445..2900
                     /label="NZF Domain"
     CDS             2445..2900
                     /label="Translation 2445-2900"
                     /translation="EGQSDERALLDQLHTLLSNTDATGLEEIDRALGIPELVNQGQALEPKQDGGWGSMAEEFVTLKDVGMDFTLGDWEQLGLEQGDTFWDTALDNCQDLFLLDPPGGGDGLGLGHEKFPSDLDLDMFNGSLECDMESIIRSELMDADGLDFNFDS"
     primer          complement(2548..2562)
                     /label="EW_NZF_R"
                     /note="sequence: GGTTCACCAGCTCGG"
     misc_feature    2607..2750
                     /label="Z (ZNF473) Activation Domain"
     misc_feature    2775..2900
                     /label="F (FOXO3) Activation Domain"
     CDS             2901..2903
                     /label="Translation 2901-2903"
                     /translation="*"
     primer          complement(2906..2927)
                     /label="EKbR"
                     /note="sequence: ttgttcGGGCCctcctcgagcg"
     CDS             2928..2954
                     /label="c-myc epitope"
     misc_feature    2954..2956
                     /label="New Stop"
     CDS             2967..2984
                     /label="Polyhistidine tag"
     misc_feature    2985..2987
                     /label="STOP (missing now)"
     primer          complement(2988..3006)
                     /label="ToFlpR"
                     /note="sequence: CTGATCAGCGGGTTTAAAC"
     polyA_signal    3006..3220
                     /label="BGH polyadenylation sequence"
     primer          complement(3007..3024)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
     primer          complement(3086..3105)
                     /label="BghR1"
                     /note="sequence: attaggaaaggacagtggga"
     primer          3197..3215
                     /label="bghF"
                     /note="sequence: AGACAATAGCAGGCATGCT"
     polyA_signal    3253..3276
                     /label="BGH pA"
     primer          complement(3355..3370)
                     /label="FlpInR"
                     /note="sequence: cgcgtaaccaccacac"
     primer          3488..3506
                     /label="FrtF"
                     /note="sequence: agggttccgatttagtgct"
     misc_feature    3560..3607
                     /label="FRT"
     gene            3615..4635
                     /label="Hygromycin"
     primer          4563..4581
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
     polyA_signal    4767..4897
                     /label="SV40 pA"
     primer          complement(4780..4805)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
     primer          complement(4804..4823)
                     /label="SV40pA-R Genewiz"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
     primer          complement(4937..4953)
                     /label="M13R Genewiz"
                     /note="sequence: CAGGAAACAGCTATGAC"
     primer          complement(4951..4970)
                     /label="M13-48REV Genewiz"
                     /note="sequence: CGGATAACAATTTCACACAG"
     primer          5229..5248
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
     rep_origin      5280..26
                     /label="pUC ori"
     primer          complement(5400..5419)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
      181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
      241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      301 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     1141 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     1201 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     1261 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     1321 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgtacgg
     1381 gccagatata cgcgttctac cgggtagggg aggcgctttt cccaaggcag tctggagcat
     1441 gcgctttagc agccccgctg ggcacttggc gctacacaag tggcctctgg cctcgcacac
     1501 attccacatc caccggtagg cgccaaccgg ctccgttctt tggtggcccc ttcgcgccac
     1561 cttctactcc tcccctagtc aggaagttcc cccccgcccc gcagctcgcg tcgtgcagga
     1621 cgtgacaaat ggaagtagca cgtctcatta gtctcgtgca gatggacagc accgctgagc
     1681 aatggaagcg ggtaggcctt tggggcagcg gccaatagca gctttgctcc ttcgctttct
     1741 gggctcagag gctgggaagg ggtgggtccg ggggcgggct caggggcggg ctcaggggcg
     1801 gggcgggcgc ccgaaggtcc tccggaggcc cggcattctg cacgcttcaa aagcgcacgt
     1861 ctgccgcgct gttctcctct tcctcatctc cgggcctttc gacctgcagc ccaagcttcg
     1921 ccaccatgTA CGAGTGTAAT GAGTGCGGTA AGACCTTCAC ACGATCTTCT AACCTTATTG
     1981 TGCATCAACG AATCCATACA GGCGAGAAGC CCTTTGCCTG CAATGACTGT GGGAAGGCTT
     2041 TTACTCAGAG TGCCAACTTG ATCGTGCATC AGAGAAGCCA TACGGGTGAG AAGCCCTATG
     2101 AATGCAAGGA ATGCGGAAAG GCCTTCTCCT GCTTCAGCCA CCTCATTGTA CACCAGCGCA
     2161 TACACACGGC TGAGAAGCCA TACGATTGCT CTGAATGTGG AAAGGCCTTT AGTCAGCTGT
     2221 CTTGCTTGAT TGTGCATCAG AGAATTCACA CGGGCGAACG CCCCTTCCAA TGCAATCAGT
     2281 GCGGGGCATC TTTCACTCAG AAGGGGAACT TACTGCGCCA TATTAAATTG CACTCTGGTG
     2341 AGAAACCGTT TAAATGTCAC TTATGCAACT ACGCCTGCAG GCGCAGAGAT GCCTTGACCG
     2401 GCCACCTCAG AACCCATggc TGGagcggag gcGATggcat gagcGAGGGC CAGAGCGACG
     2461 AGAGAGCCCT GCTGGACCAG CTGCACACCC TGCTGAGCAA CACCGACGCC ACCGGCCTGG
     2521 AGGAGATCGA CAGAGCCCTG GGCATCCCCG AGCTGGTGAA CCAGGGCCAG GCCCTGGAGC
     2581 CCAAGCAGGA CGGAGGCTGG GGATCTATGG CCGAGGAATT CGTGACCCTG AAGGACGTGG
     2641 GCATGGACTT CACCCTGGGC GACTGGGAGC AGCTGGGCCT GGAGCAGGGC GACACCTTCT
     2701 GGGACACCGC CCTGGACAAC TGCCAGGACC TGTTCCTGCT GGACCCCCCt GGCGGAGGAG
     2761 ACGGCCTTGG TTTGGGACAC GAGAAGTTCC CCAGCGACCT GGACCTGGAC ATGTTCAACG
     2821 GCAGCCTGGA GTGCGACATG GAGAGCATCA TCAGAAGCGA GCTGATGGAC GCCGACGGCC
     2881 TGGACTTCAA CTTCGACAGC taggccgctc gaggagggcc cgaacaaaaa ctcatctcag
     2941 aagaggatct gaatagcgcc gtcgaccatc atcataatca tcattgagtt taaacccgct
     3001 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3061 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3121 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3181 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggaga
     3241 tctatcgatg ttctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3301 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3361 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3421 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3481 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3541 gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta gaaagtatag
     3601 gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg
     3661 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt
     3721 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt
     3781 tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag
     3841 tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg
     3901 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg
     3961 aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg
     4021 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc
     4081 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg
     4141 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg
     4201 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact
     4261 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc
     4321 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg
     4381 caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga
     4441 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg
     4501 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct
     4561 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc
     4621 cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc ttctatgaaa
     4681 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4741 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     4801 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4861 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     4921 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     4981 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5041 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5101 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5161 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5221 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5281 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5341 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5401 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5461 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5521 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5581 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5641 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5701 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5761 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5821 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5881 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaa
//