LOCUS       EW172_CMV-TO-UTRN-14ZF- 6040 bp ds-DNA     circular     26-FEB-2025
DEFINITION  .
COMMENT     Concatenation of 4 sequences.
FEATURES             Location/Qualifiers
     promoter        4..631
                     /label="CMV"
     primer          541..561
                     /label="CMV-Forward Genewiz"
                     /note="sequence: CGCAAATGGGCGGTAGGCGTG"
     TATA_signal     576..582
                     /label="TATA box"
     misc_feature    592..610
                     /label="TetO"
     misc_feature    592..631
                     /label="TetR binding sites"
     misc_feature    613..631
                     /label="TetO"
     primer          692..712
                     /label="CMVF1"
                     /note="sequence: tgacctccatagaagacaccg"
     primer          713..728
                     /label="ToFlpF"
                     /note="sequence: GGACCGATCCAGCCTC"
     primer          735..762
                     /label="EKbF"
                     /note="sequence: ctagcgtttaaacttaagcttcgccacc"
     primer          740..761
                     /label="EWo100.1"
                     /note="sequence: gtttaaacttaagcttcgccac"
     misc_feature    755..762
                     /label="Kozak"
     misc_feature    763..765
                     /label="start codon"
     CDS             763..765
                     /label="Translation 763-765"
                     /translation="M"
     CDS             763..1431
                     /translation="MHECARCGKNFSWHSDLILHEQIHTGERPFKCYKCGRGFAEHGTLNRHLRTKGGCTGERPYNCEECGRAFSQASHLQDHQRLHTGERPHQCIECGKSFNRHCNLIRHQKIHTGERPFECSKCGKACTRRCNLIQHQKVHTGERPFACRFCAKPFRRSSDMRDHERVHTGERPFACRFCAKPFRRSSDMRDHERVHTGERPFQCELCSYTCPRRSNLDRHMKSH"
     CDS             763..1623
                     /translation="MHECARCGKNFSWHSDLILHEQIHTGERPFKCYKCGRGFAEHGTLNRHLRTKGGCTGERPYNCEECGRAFSQASHLQDHQRLHTGERPHQCIECGKSFNRHCNLIRHQKIHTGERPFECSKCGKACTRRCNLIQHQKVHTGERPFACRFCAKPFRRSSDMRDHERVHTGERPFACRFCAKPFRRSSDMRDHERVHTGERPFQCELCSYTCPRRSNLDRHMKSHAAAGGSGGSGGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGS"
     misc_feature    766..1431
                     /label="UTRN Promoter Pos(-14)-Binding ZF (Q5JNZ3 finger 1-Q66K89 finger 9-Q9NYT6 finger 19-O15535 finger 5-Q13398 finger 21-Q8NCA9 finger 4-Q8NCA9 finger 4-P49711 finger 1)"
     primer          969..985
                     /label="EWo231.1"
                     /note="sequence: TTTCTCCCAAGCTAGCC"
     primer          complement(977..994)
                     /label="EWo229.3"
                     /note="sequence: CTTGGAGATGGCTAGCTT"
     primer          1203..1220
                     /label="EWo233.1"
                     /note="sequence: TAGGTTCTGTGCTAAGCC"
     primer          complement(1213..1229)
                     /label="EWo231.4"
                     /note="sequence: CGTCTGAAAGGCTTAGC"
     primer          1430..1444
                     /label="EWo100.5"
                     /note="sequence: acgctgctgcaggcg"
     CDS             1432..1623
                     /label="Translation 1432-1623"
                     /translation="AAAGGSGGSGGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGS"
     primer          1452..1466
                     /label="EW112.2"
                     /note="sequence: aggctctggcggatc"
     primer          complement(1452..1466)
                     /label="EW112.1"
                     /note="sequence: gatccgccagagcct"
     CDS             1468..1617
                     /label="VP64"
     CDS             1624..1626
                     /label="Translation 1624-1626"
                     /translation="*"
     primer          complement(1629..1650)
                     /label="EKbR"
                     /note="sequence: ttgttcGGGCCctcctcgagcg"
     CDS             1645..1677
                     /label="c-myc epitope"
     misc_feature    1677..1679
                     /label="New Stop"
     CDS             1690..1707
                     /label="Polyhistidine tag"
     misc_feature    1708..1710
                     /label="STOP (missing now)"
     primer          complement(1711..1729)
                     /label="ToFlpR"
                     /note="sequence: CTGATCAGCGGGTTTAAAC"
     polyA_signal    1729..1943
                     /label="BGH polyadenylation sequence"
     primer          complement(1730..1747)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
     primer          complement(1809..1828)
                     /label="BghR1"
                     /note="sequence: attaggaaaggacagtggga"
     primer          1920..1938
                     /label="bghF"
                     /note="sequence: AGACAATAGCAGGCATGCT"
     polyA_signal    1976..1999
                     /label="BGH pA"
     primer          complement(2078..2093)
                     /label="FlpInR"
                     /note="sequence: cgcgtaaccaccacac"
     primer          2211..2229
                     /label="FrtF"
                     /note="sequence: agggttccgatttagtgct"
     misc_feature    2283..2330
                     /label="FRT"
     gene            2338..3358
                     /label="Hygromycin"
     primer          3286..3304
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
     polyA_signal    3490..3620
                     /label="SV40 pA"
     primer          complement(3503..3528)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
     primer          complement(3527..3546)
                     /label="SV40pA-R Genewiz"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
     primer          complement(3660..3676)
                     /label="M13R Genewiz"
                     /note="sequence: CAGGAAACAGCTATGAC"
     primer          complement(3674..3693)
                     /label="M13-48REV Genewiz"
                     /note="sequence: CGGATAACAATTTCACACAG"
     primer          3952..3971
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
     rep_origin      4003..4676
                     /label="pUC ori"
     primer          complement(4123..4142)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
     CDS             complement(4821..5681)
                     /label="Amp (bla)"
     promoter        complement(5682..5780)
                     /label="bla promoter"
     primer          5893..5913
                     /label="FlpInF1"
                     /note="sequence: agtatctgctccctgcttgtg"
     primer          5957..5974
                     /label="FlpInF"
                     /note="sequence: aacaaggcaaggcttgac"
ORIGIN
        1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctccctatca
      601 gtgatagaga tctccctatc agtgatagag atcgtcgacg agctcgttta gtgaaccgtc
      661 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      721 ccagcctccg gactctagcg tttaaactta agcttcgcca ccatgCACGA GTGTGCCCGA
      781 TGTGGGAAGA ATTTCTCTTG GCACTCAGAC CTCATCTTGC ACGAGCAAAT CCACACAGGA
      841 GAGCGACCTT TCAAGTGTTA CAAGTGTGGA CGTGGATTCG CAGAGCACGG GACACTCAAT
      901 CGACACCTTA GAACTAAGGG CGGATGTACT GGGGAGCGGC CTTACAATTG TGAGGAGTGT
      961 GGAAGAGCTT TCTCCCAAGC TAGCCATCTC CAAGACCACC AAAGGCTTCA CACTGGGGAG
     1021 AGACCCCACC AATGTATCGA GTGTGGAAAG TCCTTCAATA GACACTGTAA TCTTATAAGG
     1081 CACCAAAAGA TACACACGGG TGAGCGGCCA TTCGAGTGTT CTAAGTGTGG GAAGGCTTGT
     1141 ACTAGAAGGT GTAATTTGAT ACAACACCAA AAGGTCCACA CTGGTGAGCG TCCTTTCGCT
     1201 TGTAGGTTCT GTGCTAAGCC TTTCAGACGT TCTTCAGACA TGCGAGACCA CGAGAGAGTC
     1261 CACACTGGGG AGAGACCATT CGCATGTCGG TTCTGTGCCA AGCCTTTCAG ACGATCTTCC
     1321 GACATGCGGG ACCACGAGAG GGTTCACACA GGGGAGCGTC CCTTCCAATG TGAGCTCTGT
     1381 TCATACACTT GTCCCAGAAG GTCAAATTTA GACAGACACA TGAAGTCCCA Cgctgctgca
     1441 ggcggaagcg gaggctctgg cggatctgat gccctggacg acttcgacct ggatatgctg
     1501 ggcagcgacg ccctggatga ttttgatctg gacatgctgg gatctgacgc tctggacgat
     1561 ttcgatctcg acatgttggg atcagatgca ctggatgact ttgacctgga catgctcgga
     1621 tcataggccg ctcgaggagg gcccgaacaa aaactcatct cagaagagga tctgaatagc
     1681 gccgtcgacc atcatcataa tcatcattga gtttaaaccc gctgatcagc ctcgactgtg
     1741 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     1801 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     1861 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     1921 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg agatctatcg atgttctggg
     1981 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat
     2041 ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     2101 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     2161 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     2221 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtacc
     2281 tagaagttcc tattccgaag ttcctattct ctagaaagta taggaacttc cttggccaaa
     2341 aagcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt cgacagcgtc
     2401 tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt cgatgtagga
     2461 gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa agatcgttat
     2521 gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga cattggggaa
     2581 ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac gttgcaagac
     2641 ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat ggatgcgatc
     2701 gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca aggaatcggt
     2761 caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt gtatcactgg
     2821 caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga tgagctgatg
     2881 ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt cggctccaac
     2941 aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga ggcgatgttc
     3001 ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt ggcttgtatg
     3061 gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc gccgcggctc
     3121 cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt tgacggcaat
     3181 ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc cggagccggg
     3241 actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga tggctgtgta
     3301 gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc aaaggaatag
     3361 cacgtactac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3421 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3481 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3541 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3601 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     3661 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3721 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3781 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3841 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3901 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3961 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4021 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4081 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4141 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4201 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4261 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4321 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4381 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4441 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4501 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4561 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4621 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4681 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4741 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4801 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4861 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4921 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4981 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5041 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5101 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5161 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5221 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5281 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5341 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5401 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5461 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5521 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5581 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5641 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5701 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5761 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     5821 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     5881 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     5941 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     6001 gggttaggcg ttttgcgctg cttcgtacgg gccagatata
//