LOCUS       CMV-TO-g0_(FLP-IN)      5372 bp ds-DNA     circular     21-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
     CDS             complement(171..1031)
                     /label="Amp (bla)"
     CDS             complement(171..1031)
                     /label="Translation 1031-171"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     primer          complement(296..314)
                     /label="EKo152.2"
                     /note="sequence: tctcgcggtatcattgcag"
     promoter        complement(1032..1130)
                     /label="bla promoter"
     primer          1126..1142
                     /label="AmpR_seq_F"
                     /note="sequence: ggggttccgcgcacatt"
     primer          1243..1263
                     /label="FlpInF1"
                     /note="sequence: agtatctgctccctgcttgtg"
     primer          1307..1324
                     /label="FlpInF"
                     /note="sequence: aacaaggcaaggcttgac"
     primer          1318..1337
                     /label="FlpInF2"
                     /note="sequence: gcttgaccgacaattgcatg"
     primer          complement(1322..1344)
                     /label="EKo152.4"
                     /note="sequence: gattcttcatgcaattgtcggtc"
     primer          1378..1394
                     /label="EKo279.1"
                     /note="sequence: cgggccagatatacgcg"
     primer          1379..1425
                     /label="EKo272.1"
                     /note="sequence: gggccagatatacgcgttGACATTGATTATTGACTAGTTATTAATAG"
     promoter        1394..2021
                     /label="CMV"
     enhancer        1394..2120
                     /label="CMV promoter (with TetOs)"
     primer          1473..1495
                     /label="EK_CMVFa"
                     /note="sequence: cgttacataacttacggtaaatg"
     primer          1549..1568
                     /label="EK_CMVFb"
                     /note="sequence: tatgttcccatagtaacgcc"
     primer          1625..1645
                     /label="EK_CMVFc"
                     /note="sequence: tggcagtacatcaagtgtatc"
     primer          1701..1719
                     /label="EK_CMVFd"
                     /note="sequence: ttatgcccagtacatgacc"
     primer          1777..1794
                     /label="EK_CMVFe"
                     /note="sequence: gtgatgcggttttggcag"
     misc_feature    1879..1879
                     /label="A"
     primer          1931..1951
                     /label="CMV-forward Genewiz"
                     /note="sequence: CGCAAATGGGCGGTAGGCGTG"
     primer          1931..1951
                     /label="CMV-Forward Genewiz"
                     /note="sequence: CGCAAATGGGCGGTAGGCGTG"
     TATA_signal     1966..1972
                     /label="TATA box"
     protein_binding 1982..2000
                     /label="TetO"
     protein_binding 1982..2021
                     /label="TetR binding sites"
     protein_binding 2003..2021
                     /label="TetO"
     primer          2082..2102
                     /label="CMVF1"
                     /note="sequence: tgacctccatagaagacaccg"
     primer          2103..2118
                     /label="ToFlpF"
                     /note="sequence: GGACCGATCCAGCCTC"
     primer          complement(2130..2144)
                     /label="EKo283.2"
                     /note="sequence: agcttaagtttaaac"
     misc_feature    2146..2188
                     /label="HH"
     misc_feature    2189..2203
                     /label="g0"
     misc_feature    2220..2283
                     /label="tracrRNA"
     misc_feature    2284..2351
                     /label="HDV"
     primer          complement(2351..2372)
                     /label="EKbR"
                     /note="sequence: ttgttcGGGCCctcctcgagcg"
     primer          complement(2355..2372)
                     /label="pNSK053"
                     /note="sequence: ttgttcgggccctcctcg"
     primer          complement(2362..2383)
                     /label="pNK018"
                     /note="sequence: gagatgagtttttgttcGGGCC"
     CDS             2367..2399
                     /label="c-myc epitope"
     misc_feature    2399..2401
                     /label="New Stop"
     CDS             2412..2429
                     /label="Polyhistidine tag"
     misc_feature    2421..2421
                     /label="c -> a"
     misc_feature    2430..2432
                     /label="cya"
     primer          complement(2433..2451)
                     /label="ToFlpR"
                     /note="sequence: CTGATCAGCGGGTTTAAAC"
     polyA_signal    2451..2665
                     /label="BGH polyadenylation sequence"
     primer          complement(2452..2469)
                     /label="BGHR Genewiz"
                     /note="sequence: TAGAAGGCACAGTCGAGG"
     primer          complement(2531..2550)
                     /label="BghR1"
                     /note="sequence: attaggaaaggacagtggga"
     primer          2642..2660
                     /label="bghF"
                     /note="sequence: AGACAATAGCAGGCATGCT"
     primer          complement(2645..2662)
                     /label="EKo326.3"
                     /note="sequence: ccagcatgcctgctattg"
     polyA_signal    2698..2721
                     /label="BGH pA"
     primer          2722..2740
                     /label="EKo293.3"
                     /note="sequence: cttctgaggcggaaagaac"
     primer          complement(2800..2815)
                     /label="FlpInR"
                     /note="sequence: cgcgtaaccaccacac"
     misc_feature    2925..2925
                     /label="C insertion"
     primer          2933..2951
                     /label="FrtF"
                     /note="sequence: agggttccgatttagtgct"
     misc_feature    3005..3052
                     /label="FRT"
     primer          complement(3026..3053)
                     /label="EKo299.3"
                     /note="sequence: ggaagttcctatactttctagagaatag"
     gene            3060..4080
                     /label="Hygromycin"
     primer          3288..3309
                     /label="EKo152.1"
                     /note="sequence: cttgacattggggaattcagcg"
     primer          4008..4026
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
     polyA_signal    4212..4342
                     /label="SV40 pA"
     primer          complement(4225..4250)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
     primer          complement(4249..4268)
                     /label="SV40pA-R Genewiz"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
     primer          complement(4382..4398)
                     /label="M13R Genewiz"
                     /note="sequence: CAGGAAACAGCTATGAC"
     primer          complement(4396..4415)
                     /label="M13-48REV Genewiz"
                     /note="sequence: CGGATAACAATTTCACACAG"
     primer          4674..4693
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
     rep_origin      4725..26
                     /label="pUC ori"
     primer          complement(4845..4864)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
     misc_feature    5225..5225
                     /label="g -> a"
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 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
     1441 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     1501 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     1561 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     1621 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
     1681 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
     1741 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
     1801 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     1861 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
     1921 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     1981 ctccctatca gtgatagaga tctccctatc agtgatagag atcgtcgacg agctcgttta
     2041 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     2101 cgggaccgat ccagcctccg gactctagcg tttaaactta agcttGTGGA CCTGATGAGT
     2161 CCGTGAGGAC GAAACGAGTA AGCTCGTCGT CCACCCCACA GTGgttttag agctaGAAAt
     2221 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgcT
     2281 TTTGGCCGGC ATGGTCCCAG CCTCCTCGCT GGCGCCGGCT GGGCAACATG CTTCGGCATG
     2341 GCGAATGGGA Cgctcgagga gggcccgaac aaaaactcat ctcagaagag gatctgaata
     2401 gcgccgtcga ccatcatcat aatcatcatt gagtttaaac ccgctgatca gcctcgactg
     2461 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2521 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2581 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2641 aagacaatag caggcatgct ggggatgcgg tgggctctat ggagatctat cgatgttctg
     2701 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     2761 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2821 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2881 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2941 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3001 cctagaagtt cctattccga agttcctatt ctctagaaag tataggaact tccttggcca
     3061 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
     3121 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
     3181 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
     3241 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
     3301 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
     3361 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga
     3421 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
     3481 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
     3541 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
     3601 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
     3661 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
     3721 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
     3781 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc
     3841 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
     3901 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
     3961 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
     4021 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat
     4081 agcacgtact acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4141 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4201 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     4261 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     4321 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
     4381 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     4441 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     4501 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     4561 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     4621 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4681 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4741 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4801 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4861 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4921 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4981 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5041 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5101 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     5161 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     5221 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5281 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5341 agcagattac gcgcagaaaa aaaggatctc aa
//