LOCUS       EW434_4x(ZNF35/ZNF35tar 5784 bp ds-DNA     circular     27-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          10..27
                     /label="FlpInF"
                     /note="sequence: aacaaggcaaggcttgac"
     primer_bind     21..31
                     /label="primer3"
     primer          32..105
                     /label="EWo107.1"
                     /note="sequence: aattgacgattgagtaacactgtatctagtctacctacagggctcATATATAAAGAGATATATAAAGAGctctt"
     primer          complement(36..101)
                     /label="EWo103.2"
                     /note="sequence: gCTCTTTATATATCTCTTTATATATgagccctgtaggtagactagatacagtgttactcaatcgtc"
     misc_feature    77..100
                     /label="ZNF35/ZNF35 site"
     primer          complement(102..201)
                     /label="EWo145.2"
                     /note="sequence: ctagactgataCTCTTTATATATCTCTTTATATATgttctgCTCTTTATATATCTCTTTATATATgacctcCTCTTTATATATCTCTTTATATATgaaga"
     primer          106..197
                     /label="EWo145.1"
                     /note="sequence: cATATATAAAGAGATATATAAAGAGgaggtcATATATAAAGAGATATATAAAGAGcagaacATATATAAAGAGATATATAAAGAGtatcagt"
     misc_feature    107..130
                     /label="ZNF35/ZNF35 site"
     misc_feature    137..160
                     /label="ZNF35/ZNF35 site"
     misc_feature    167..190
                     /label="ZNF35/ZNF35 site"
     misc_feature    227..718
                     /label="SFFV"
     primer          637..653
                     /label="EK_SFFV_F2"
                     /note="sequence: gcttctgcttcccgagc"
     primer          709..734
                     /label="EK_SFFV_F"
                     /note="sequence: gtcgcccgggaagcttcgccaccatg"
     primer          719..734
                     /label="EKo271.3"
                     /note="sequence: aagcttcgccaccatg"
     primer          720..735
                     /label="EKo309.1"
                     /note="sequence: agcttcgccaccatgg"
     primer          complement(721..739)
                     /label="EKo237.2"
                     /note="sequence: gatcccatggtggcgaagc"
     misc_feature    724..731
                     /label="Kozak"
     primer          complement(724..739)
                     /label="EKo309.2"
                     /note="sequence: gatcccatggtggcga"
     CDS             732..1454
                     /label="GFP"
     CDS             732..1463
                     /label="Translation 732-1463"
                     /translation="MGSVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKTG*"
     misc_feature    735..740
                     /label="for cloning"
     primer          741..759
                     /label="EKo147.1"
                     /note="sequence: GTCTCAAAGGGCGAGGAAC"
     primer          complement(774..794)
                     /label="GFP5"
                     /note="sequence: CAATTCGACGAGGATCGGAAC"
     primer          complement(997..1013)
                     /label="EKo224.2"
                     /note="sequence: ACCTTCAGGCATAGCGG"
     primer          1105..1125
                     /label="pPYK028"
                     /note="sequence: GGATCGAACTGAAGGGCATAG"
     primer          1382..1401
                     /label="GFP6"
                     /note="sequence: ACGGGATCACATGGTCCTTC"
     primer          complement(1433..1487)
                     /label="EKo25.2"
                     /note="sequence: ttgttcgggccctcctcgagcggcctaAccggtCTTATACAGTTCGTCCATCCCG"
     misc_feature    1455..1460
                     /label="for cloning"
     primer          1456..1480
                     /label="EKo130.1"
                     /note="sequence: ccggTtaggccgctcgaggagggcc"
     primer          complement(1460..1476)
                     /label="EKo130.2"
                     /note="sequence: ctcctcgagcggcctaA"
     CDS             1461..1462
                     /label="Translation 1461-1462"
                     /translation=""
     primer          complement(1466..1487)
                     /label="EKbR"
                     /note="sequence: ttgttcGGGCCctcctcgagcg"
     primer          complement(1477..1498)
                     /label="pNK018"
                     /note="sequence: gagatgagtttttgttcGGGCC"
     CDS             1482..1514
                     /label="c-myc epitope"
     misc_feature    1514..1516
                     /label="New Stop"
     CDS             1527..1544
                     /label="Polyhistidine tag"
     misc_feature    1545..1547
                     /label="cya"
     primer          complement(1548..1566)
                     /label="ToFlpR"
                     /note="sequence: CTGATCAGCGGGTTTAAAC"
     polyA_signal    1566..1780
                     /label="BGH polyadenylation sequence"
     primer          complement(1567..1584)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
     primer          complement(1646..1665)
                     /label="BghR1"
                     /note="sequence: attaggaaaggacagtggga"
     primer          1757..1775
                     /label="bghF"
                     /note="sequence: AGACAATAGCAGGCATGCT"
     polyA_signal    1813..1836
                     /label="BGH pA"
     primer          complement(1915..1930)
                     /label="FlpInR"
                     /note="sequence: cgcgtaaccaccacac"
     primer          2048..2066
                     /label="FrtF"
                     /note="sequence: agggttccgatttagtgct"
     misc_feature    2120..2167
                     /label="FRT"
     gene            2175..3195
                     /label="Hygromycin"
     primer          2403..2424
                     /label="EKo152.1"
                     /note="sequence: cttgacattggggaattcagcg"
     primer          3123..3141
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
     polyA_signal    3327..3457
                     /label="SV40 pA"
     primer          complement(3340..3365)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
     primer          complement(3364..3383)
                     /label="SV40pA-R Genewiz"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
     primer          complement(3497..3513)
                     /label="M13R Genewiz"
                     /note="sequence: CAGGAAACAGCTATGAC"
     primer          complement(3511..3530)
                     /label="M13-48REV Genewiz"
                     /note="sequence: CGGATAACAATTTCACACAG"
     primer          3789..3808
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
     rep_origin      3840..4513
                     /label="pUC ori"
     primer          complement(3960..3979)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
     CDS             complement(4658..5518)
                     /label="Amp (bla)"
     primer          complement(4783..4801)
                     /label="EKo152.2"
                     /note="sequence: tctcgcggtatcattgcag"
     promoter        complement(5519..5617)
                     /label="bla promoter"
     primer          5730..5750
                     /label="FlpInF1"
                     /note="sequence: agtatctgctccctgcttgtg"
ORIGIN
        1 ttaagctaca acaaggcaag gcttgaccga caattgacga ttgagtaaca ctgtatctag
       61 tctacctaca gggctcATAT ATAAAGAGAT ATATAAAGAG ctcttcATAT ATAAAGAGAT
      121 ATATAAAGAG gaggtcATAT ATAAAGAGAT ATATAAAGAG cagaacATAT ATAAAGAGAT
      181 ATATAAAGAG tatcagtcta gagggtatat aatggatccc aaattgccga taaaataaaa
      241 gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa
      301 gctagctgca gtaacgccat tttgcaaggc atggaaaaat accaaaccaa gaatagagaa
      361 gttcagatca agggcgggta catgaaaata gctaacgttg ggccaaacag gatatctgcg
      421 gtgagcagtt tcggccccgg cccggggcca agaacagatg gtcaccgcag tttcggcccc
      481 ggcccgaggc caagaacaga tggtccccag atatggccca accctcagca gtttcttaag
      541 acccatcaga tgtttccagg ctcccccaag gacctgaaat gaccctgcgc cttatttgaa
      601 ttaaccaatc agcctgcttc tcgcttctgt tcgcgcgctt ctgcttcccg agctctataa
      661 aagagctcac aacccctcac tcggcgcgcc agtcctccga cagactgagt cgcccgggaa
      721 gcttcgccac cATGggatcc GTCTCAAAGG GCGAGGAACT TTTTACGGGC GTTGTTCCGA
      781 TCCTCGTCGA ATTGGACGGG GACGTTAATG GGCATAAATT CTCAGTTAGT GGTGAGGGTG
      841 AGGGGGACGC GACATATGGG AAACTTACGC TGAAGTTCAT ATGTACCACC GGAAAATTGC
      901 CAGTGCCGTG GCCTACTCTC GTCACCACCC TGACATATGG TGTTCAGTGT TTTAGCCGGT
      961 ATCCAGACCA TATGAAACAG CACGATTTTT TTAAATCCGC TATGCCTGAA GGTTACGTAC
     1021 AGGAGCGAAC GATATTTTTT AAAGACGACG GAAACTACAA AACTAGAGCC GAGGTAAAAT
     1081 TCGAGGGTGA CACTCTCGTG AACCGGATCG AACTGAAGGG CATAGATTTT AAGGAGGATG
     1141 GCAATATACT GGGCCACAAG CTCGAATACA ACTATAATTC TCATAATGTC TATATCATGG
     1201 CGGACAAACA GAAAAACGGC ATCAAGGTAA ACTTTAAAAT CCGACACAAT ATTGAGGACG
     1261 GCTCTGTCCA GCTTGCGGAC CATTACCAAC AGAACACGCC GATAGGGGAT GGTCCAGTAC
     1321 TTCTCCCTGA TAATCACTAT TTGTCTACTC AGTCAGCTCT TAGCAAGGAC CCAAACGAAA
     1381 AACGGGATCA CATGGTCCTT CTTGAGTTCG TCACTGCGGC TGGGATAACA CTCGGGATGG
     1441 ACGAACTGTA TAAGaccggT taggccgctc gaggagggcc cgaacaaaaa ctcatctcag
     1501 aagaggatct gaatagcgcc gtcgaccatc atcataatca tcattgagtt taaacccgct
     1561 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     1621 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     1681 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     1741 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggaga
     1801 tctatcgatg ttctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     1861 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     1921 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     1981 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     2041 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     2101 gtgatggttc acgtacctag aagttcctat tccgaagttc ctattctcta gaaagtatag
     2161 gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg
     2221 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt
     2281 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt
     2341 tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag
     2401 tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg
     2461 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg
     2521 aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg
     2581 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc
     2641 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg
     2701 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg
     2761 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact
     2821 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc
     2881 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg
     2941 caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga
     3001 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg
     3061 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct
     3121 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc
     3181 cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc ttctatgaaa
     3241 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3301 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     3361 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3421 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     3481 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3541 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3601 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3661 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3721 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3781 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3841 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3901 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3961 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4021 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4081 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4141 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4201 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4261 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4321 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4381 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4441 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4501 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4561 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4621 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4681 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4741 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4801 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4861 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4921 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4981 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5041 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5101 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5161 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5221 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5281 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5341 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5401 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5461 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5521 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5581 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5641 tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac
     5701 aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt
     5761 cgctgagtag tgcgcgagca aaat
//