LOCUS       EW425_CMV-TO-ZNF35(5-8) 5830 bp ds-DNA     circular     26-FEB-2025
DEFINITION  .
COMMENT     Concatenation of 4 sequences.
FEATURES             Location/Qualifiers
     promoter        1..628
                     /label="CMV"
     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          728..762
                     /label="EWo300"
                     /note="sequence: gactctagcgtttaaacttaagcttcgccaccatg"
     primer          732..759
                     /label="EKbF"
                     /note="sequence: ctagcgtttaaacttaagcttcgccacc"
     primer          737..758
                     /label="EWo100.1"
                     /note="sequence: gtttaaacttaagcttcgccac"
     misc_feature    752..759
                     /label="Kozak"
     misc_feature    760..762
                     /label="start codon"
     CDS             760..762
                     /label="Translation 760-762"
                     /translation="M"
     misc_feature    763..1083
                     /label="ZNF35 ZF Domains 5-8"
     CDS             763..1098
                     /label="Translation 763-1098"
                     /translation="YECNECGKTFTRSSNLIVHQRIHTGEKPFACNDCGKAFTQSANLIVHQRSHTGEKPYECKECGKAFSCFSHLIVHQRIHTAEKPYDCSECGKAFSQLSCLIVHQRIHTGERP"
     primer          complement(815..846)
                     /label="EWo153.1"
                     /note="sequence: GGGCTTCTCGCCTGTATGGATTCGTTGATGCA"
     primer          970..987
                     /label="EWo51"
                     /note="sequence: AGCCACCTCATTGTACAC"
     primer          complement(980..996)
                     /label="EWo126"
                     /note="sequence: TATGCGCTGGTGTACAA"
     primer          complement(1064..1083)
                     /label="EWo169.1"
                     /note="sequence: GTGAATTCTCTGATGCACAA"
     misc_feature    1084..1098
                     /label="Canonical ZF Linker"
     primer          complement(1084..1098)
                     /label="EWo120.1"
                     /note="sequence: GGGGCGTTCGCCCGT"
     misc_feature    1099..1251
                     /label="IKZF1 ZF 2-4"
     CDS             1099..1251
                     /label="Translation 1099-1251"
                     /translation="FQCNQCGASFTQKGNLLRHIKLHSGEKPFKCHLCNYACRRRDALTGHLRTH"
     primer          complement(1235..1251)
                     /label="EWo366.1"
                     /note="sequence: ATGGGTTCTGAGGTGGC"
     CDS             1252..1278
                     /label="Translation 1252-1278"
                     /translation="GWSGGDGMS"
     primer          complement(1264..1278)
                     /label="EWo425.1"
                     /note="sequence: gctcatgccATCgcc"
     primer          1264..1295
                     /label="EWo425.2"
                     /note="sequence: ggcGATggcatgagcTGCGTGACCTTTGAAGA"
     misc_feature    1279..1401
                     /label="ZIK1 KRAB Domain"
     CDS             1279..1404
                     /label="Translation 1279-1404"
                     /translation="CVTFEDIAIYFSQDEWGLLDEAQRLLYLEVMLENFALVASL*"
     primer          complement(1387..1437)
                     /label="EWo416.2"
                     /note="sequence: ttgttcgggccctcctcgagcggcctatgatccctaCAGGGATGCCACGAG"
     primer          complement(1416..1437)
                     /label="EKbR"
                     /note="sequence: ttgttcGGGCCctcctcgagcg"
     primer          complement(1420..1437)
                     /label="pNSK053"
                     /note="sequence: ttgttcgggccctcctcg"
     CDS             1432..1464
                     /label="c-myc epitope"
     misc_feature    1464..1466
                     /label="New Stop"
     CDS             1477..1494
                     /label="Polyhistidine tag"
     misc_feature    1495..1497
                     /label="STOP (missing now)"
     primer          complement(1498..1516)
                     /label="ToFlpR"
                     /note="sequence: CTGATCAGCGGGTTTAAAC"
     polyA_signal    1516..1730
                     /label="BGH polyadenylation sequence"
     primer          complement(1517..1534)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
     primer          complement(1596..1615)
                     /label="BghR1"
                     /note="sequence: attaggaaaggacagtggga"
     primer          1707..1725
                     /label="bghF"
                     /note="sequence: AGACAATAGCAGGCATGCT"
     polyA_signal    1763..1786
                     /label="BGH pA"
     primer          complement(1865..1880)
                     /label="FlpInR"
                     /note="sequence: cgcgtaaccaccacac"
     primer          1998..2016
                     /label="FrtF"
                     /note="sequence: agggttccgatttagtgct"
     misc_feature    2070..2117
                     /label="FRT"
     gene            2125..3145
                     /label="Hygromycin"
     primer          3073..3091
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
     polyA_signal    3277..3407
                     /label="SV40 pA"
     primer          complement(3290..3315)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
     primer          complement(3314..3333)
                     /label="SV40pA-R Genewiz"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
     primer          complement(3447..3463)
                     /label="M13R Genewiz"
                     /note="sequence: CAGGAAACAGCTATGAC"
     primer          complement(3461..3480)
                     /label="M13-48REV Genewiz"
                     /note="sequence: CGGATAACAATTTCACACAG"
     primer          3739..3758
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
     rep_origin      3790..4463
                     /label="pUC ori"
     primer          complement(3910..3929)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
     CDS             complement(4608..5468)
                     /label="Amp (bla)"
     promoter        complement(5469..5567)
                     /label="bla promoter"
     primer          5680..5700
                     /label="FlpInF1"
                     /note="sequence: agtatctgctccctgcttgtg"
     primer          5744..5761
                     /label="FlpInF"
                     /note="sequence: aacaaggcaaggcttgac"
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 AACGCCCCTT CCAATGCAAT CAGTGCGGGG CATCTTTCAC TCAGAAGGGG
     1141 AACTTACTGC GCCATATTAA ATTGCACTCT GGTGAGAAAC CGTTTAAATG TCACTTATGC
     1201 AACTACGCCT GCAGGCGCAG AGATGCCTTG ACCGGCCACC TCAGAACCCA TggcTGGagc
     1261 ggaggcGATg gcatgagcTG CGTGACCTTT GAAGACATTG CCATCTATTT TAGTCAAGAC
     1321 GAATGGGGGC TCCTGGACGA AGCCCAGAGA CTTCTGTACC TGGAGGTGAT GCTTGAAAAC
     1381 TTCGCTCTCG TGGCATCCCT Gtagggatca taggccgctc gaggagggcc cgaacaaaaa
     1441 ctcatctcag aagaggatct gaatagcgcc gtcgaccatc atcataatca tcattgagtt
     1501 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1561 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1621 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     1681 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     1741 tctatggaga tctatcgatg ttctggggat gcggtgggct ctatggcttc tgaggcggaa
     1801 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     1861 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     1921 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     1981 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2041 cttgattagg gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta
     2101 gaaagtatag gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag
     2161 tttctgatcg aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa
     2221 tctcgtgctt tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc
     2281 gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg
     2341 attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc
     2401 cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag
     2461 ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc
     2521 ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg
     2581 attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc
     2641 gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac
     2701 ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg
     2761 gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc
     2821 ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat
     2881 ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa
     2941 ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc
     3001 gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc
     3061 gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc
     3121 agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc
     3181 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     3241 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     3301 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3361 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     3421 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3481 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3541 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3601 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3661 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3721 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3781 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3841 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     3901 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     3961 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4021 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4081 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4141 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4201 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4261 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4321 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4381 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4441 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     4501 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     4561 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     4621 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     4681 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     4741 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     4801 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     4861 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     4921 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     4981 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     5041 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     5101 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     5161 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     5221 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     5281 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     5341 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     5401 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     5461 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     5521 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     5581 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     5641 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     5701 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     5761 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgtacgggcc
     5821 agatatacgc
//