LOCUS       NSK177_U6TO_EBER2       4243 bp ds-DNA     circular     07-JAN-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          113..133
                     /label="FlpInF1"
                     /note="sequence: agtatctgctccctgcttgtg"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          177..194
                     /label="FlpInF"
                     /note="sequence: aacaaggcaaggcttgac"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          188..207
                     /label="FlpInF2"
                     /note="sequence: gcttgaccgacaattgcatg"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer          complement(192..214)
                     /label="EKo152.4"
                     /note="sequence: gattcttcatgcaattgtcggtc"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          248..264
                     /label="EKo279.1"
                     /note="sequence: cgggccagatatacgcg"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer          250..285
                     /label="EKo251.1"
                     /note="sequence: ggccagatatacgcgtgtgagggcctatttcccatg"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     promoter        277..528
                     /label="U6"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          439..458
                     /label="U6 Genewiz"
                     /note="sequence: GACTATCATATGCTTACCGT"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer          complement(439..461)
                     /label="EKo243.1"
                     /note="sequence: gttacggtaagcatatgatagtc"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer_bind     440..461
                     /label="LKO"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          440..461
                     /label="U6F"
                     /note="sequence: actatcatatgcttaccgtaac"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     protein_binding 469..487
                     /label="TetO"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     protein_binding 495..513
                     /label="TetO"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          complement(517..536)
                     /label="EKo299.1"
                     /note="sequence: CGGTGTTTCGTCCTTTCCAC"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    529..535
                     /label="fixed"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature    532..535
                     /label="overhang"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     CDS             535..535
                     /label="Translation 535-535"
                     /translation=""
     misc_feature    536..536
                     /label="required first G?"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    537..542
                     /label="extra for cloning"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer          538..629
                     /label="pNSK451"
                     /note="sequence: tcgagAGGACAGCCGTTGCCCTAGTGGTTTCGGACACACCGCCAACGCTCAGTGCGGTGCTACCGACCCGAGGTCAAGTCCCGGGGGAGGAG"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          complement(542..625)
                     /label="pNSK452"
                     /note="sequence: TCCCCCGGGACTTGACCTCGGGTCGGTAGCACCGCACTGAGCGTTGGCGGTGTGTCCGAAACCACTAGGGCAACGGCTGTCCTc"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    543..632
                     /label="os1"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    543..715
                     /label="EBER2 consensus"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    553..642
                     /label="os2"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    563..652
                     /label="os3"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature    573..662
                     /label="os4"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    579..581
                     /label="CCA"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    583..672
                     /label="os5"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    593..682
                     /label="os6"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    603..692
                     /label="os7"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    613..702
                     /label="os8"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    623..712
                     /label="os9"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    626..715
                     /label="os10"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     primer          complement(626..716)
                     /label="pNSK454"
                     /note="sequence: cAAAAACAGCGGACAAGCCGAATACCCTTCTCCCAGAGGGATTAGAGAATCCTGACTTGCAAATGCTCTAGGCGGGAAGCCTCTCTTCTCC"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          630..720
                     /label="pNSK453"
                     /note="sequence: AAGAGAGGCTTCCCGCCTAGAGCATTTGCAAGTCAGGATTCTCTAATCCCTCTGGGAGAAGGGTATTCGGCTTGTCCGCTGTTTTTgggcc"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    716..721
                     /label="extra for cloning"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(721..777)
                     /label="EKo298.4"
                     /note="sequence: ctagagaataggaacttcggaataggaacttctagggggttcctgcggccAAAAAAg"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             722..722
                     /label="Translation 722-722"
                     /translation=""
     misc_feature    722..728
                     /label="U6 term"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    complement(728..731)
                     /label="overhang"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(728..742)
                     /label="EKo243.2"
                     /note="sequence: ggggttcctgcggcc"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          complement(728..794)
                     /label="EKo251.2"
                     /note="sequence: ggaagttcctatactttctagagaataggaacttcggaataggaacttctagggggttcctgcggcc"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    746..793
                     /label="FRT"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          complement(767..794)
                     /label="EKo299.3"
                     /note="sequence: ggaagttcctatactttctagagaatag"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     gene            801..1821
                     /label="Hygromycin"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          1029..1050
                     /label="EKo152.1"
                     /note="sequence: cttgacattggggaattcagcg"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer          1749..1767
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     polyA_signal    1953..2083
                     /label="SV40 pA"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          complement(1966..1991)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          complement(1990..2009)
                     /label="SV40pA-R Genewiz"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          complement(2123..2139)
                     /label="M13R Genewiz"
                     /note="sequence: CAGGAAACAGCTATGAC"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(2137..2156)
                     /label="M13-48REV Genewiz"
                     /note="sequence: CGGATAACAATTTCACACAG"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          2415..2434
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     rep_origin      2466..3139
                     /label="pUC ori"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          complement(2586..2605)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     CDS             complement(3284..4144)
                     /label="Amp (bla)"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     CDS             complement(3284..4144)
                     /label="Translation 4144-3284"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     primer          complement(3409..3427)
                     /label="EKo152.2"
                     /note="sequence: tctcgcggtatcattgcag"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     promoter        complement(4145..4243)
                     /label="bla promoter"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          4239..12
                     /label="AmpR_seq_F"
                     /note="sequence: ggggttccgcgcacatt"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
ORIGIN
        1 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
       61 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
      121 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
      181 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
      241 cttcgtacgg gccagatata cgcgtgtgag ggcctatttc ccatgattcc ttcatatttg
      301 catatacgat acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag
      361 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
      421 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagtc cctatcagtg
      481 atagagatat atattcccta tcagtgatag agacttGTGG AAAGGACGAA ACACCGctcg
      541 agAGGACAGC CGTTGCCCTA GTGGTTTCGG ACACACCGCC AACGCTCAGT GCGGTGCTAC
      601 CGACCCGAGG TCAAGTCCCG GGGGAGGAGA AGAGAGGCTT CCCGCCTAGA GCATTTGCAA
      661 GTCAGGATTC TCTAATCCCT CTGGGAGAAG GGTATTCGGC TTGTCCGCTG TTTTTgggcc
      721 cTTTTTTggc cgcaggaacc ccctagaagt tcctattccg aagttcctat tctctagaaa
      781 gtataggaac ttccttggcc aaaaagcctg aactcaccgc gacgtctgtc gagaagtttc
      841 tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc gaagaatctc
      901 gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat agctgcgccg
      961 atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg ctcccgattc
     1021 cggaagtgct tgacattggg gaattcagcg agagcctgac ctattgcatc tcccgccgtg
     1081 cacagggtgt cacgttgcaa gacctgcctg aaaccgaact gcccgctgtt ctgcagccgg
     1141 tcgcggaggc catggatgcg atcgctgcgg ccgatcttag ccagacgagc gggttcggcc
     1201 cattcggacc gcaaggaatc ggtcaataca ctacatggcg tgatttcata tgcgcgattg
     1261 ctgatcccca tgtgtatcac tggcaaactg tgatggacga caccgtcagt gcgtccgtcg
     1321 cgcaggctct cgatgagctg atgctttggg ccgaggactg ccccgaagtc cggcacctcg
     1381 tgcacgcgga tttcggctcc aacaatgtcc tgacggacaa tggccgcata acagcggtca
     1441 ttgactggag cgaggcgatg ttcggggatt cccaatacga ggtcgccaac atcttcttct
     1501 ggaggccgtg gttggcttgt atggagcagc agacgcgcta cttcgagcgg aggcatccgg
     1561 agcttgcagg atcgccgcgg ctccgggcgt atatgctccg cattggtctt gaccaactct
     1621 atcagagctt ggttgacggc aatttcgatg atgcagcttg ggcgcagggt cgatgcgacg
     1681 caatcgtccg atccggagcc gggactgtcg ggcgtacaca aatcgcccgc agaagcgcgg
     1741 ccgtctggac cgatggctgt gtagaagtac tcgccgatag tggaaaccga cgccccagca
     1801 ctcgtccgag ggcaaaggaa tagcacgtac tacgagattt cgattccacc gccgccttct
     1861 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     1921 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     1981 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2041 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     2101 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     2161 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     2221 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     2281 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     2341 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2401 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2461 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2521 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2581 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2641 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2701 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2761 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2821 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2881 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2941 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     3001 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     3061 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     3121 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     3181 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     3241 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     3301 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     3361 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     3421 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     3481 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     3541 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     3601 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     3661 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     3721 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     3781 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     3841 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     3901 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     3961 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     4021 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     4081 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     4141 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     4201 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggt
//