LOCUS       NSK176_U6TO_EBER1       4237 bp ds-DNA     circular     25-NOV-2023
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"
     misc_feature    277..721
                     /label="insert"
                     /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..625
                     /label="pNSK447"
                     /note="sequence: tcgagAGGACCTACGCTGCCCTAGAGGTTTTGCTAGGGAGGAGACGTGTGTGGCTGTAGCCACCCGTCCCGGGTACAAGTCCCGGGTG"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          complement(542..621)
                     /label="pNSK448"
                     /note="sequence: CGGGACTTGTACCCGGGACGGGTGGCTACAGCCACACACGTCTCCTCCCTAGCAAAACCTCTAGGGCAGCGTAGGTCCTc"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    543..632
                     /label="os1"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    543..709
                     /label="EBER1 consensus"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    553..642
                     /label="os2"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    555..644
                     /label="linear 1"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    563..652
                     /label="os3"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    573..662
                     /label="os4"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    583..672
                     /label="os5"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    593..682
                     /label="os6"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     misc_feature    597..599
                     /label="CCA"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    603..692
                     /label="os7"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    611..700
                     /label="linear 2"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    613..702
                     /label="os8"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    620..709
                     /label="os9"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          complement(622..710)
                     /label="pNSK450"
                     /note="sequence: cAAAACATGCGGACCACCAGCTGGTACTTGACCGAAGACGGCAGAAAGCAGAGTCTGGGAAGACAACCACAGACACCGTCCTCACCACC"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          626..714
                     /label="pNSK449"
                     /note="sequence: GTGAGGACGGTGTCTGTGGTTGTCTTCCCAGACTCTGCTTTCTGCCGTCTTCGGTCAAGTACCAGCTGGTGGTCCGCATGTTTTgggcc"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    653..655
                     /label="CCA"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    710..715
                     /label="extra for cloning"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          complement(715..771)
                     /label="EKo298.4"
                     /note="sequence: ctagagaataggaacttcggaataggaacttctagggggttcctgcggccAAAAAAg"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             716..716
                     /label="Translation 716-716"
                     /translation=""
     misc_feature    716..721
                     /label="U6 term"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     misc_feature    complement(722..725)
                     /label="overhang"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(722..736)
                     /label="EKo243.2"
                     /note="sequence: ggggttcctgcggcc"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          complement(722..788)
                     /label="EKo251.2"
                     /note="sequence: ggaagttcctatactttctagagaataggaacttcggaataggaacttctagggggttcctgcggcc"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    740..787
                     /label="FRT"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          complement(761..788)
                     /label="EKo299.3"
                     /note="sequence: ggaagttcctatactttctagagaatag"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     gene            795..1815
                     /label="Hygromycin"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          1023..1044
                     /label="EKo152.1"
                     /note="sequence: cttgacattggggaattcagcg"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer          1743..1761
                     /label="Hyg_F"
                     /note="sequence: ACCGATGGCTGTGTAGAAG"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     polyA_signal    1947..2077
                     /label="SV40 pA"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          complement(1960..1985)
                     /label="SV40PA_R"
                     /note="sequence: GCTTTATTTGTAACCATTATAAGCTG"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer          complement(1984..2003)
                     /label="SV40pA-R Genewiz"
                     /note="sequence: GAAATTTGTGATGCTATTGC"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     primer          complement(2117..2133)
                     /label="M13R Genewiz"
                     /note="sequence: CAGGAAACAGCTATGAC"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(2131..2150)
                     /label="M13-48REV Genewiz"
                     /note="sequence: CGGATAACAATTTCACACAG"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          2409..2428
                     /label="pUCori5F"
                     /note="sequence: AAGGCGGTAATACGGTTATC"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     rep_origin      2460..3133
                     /label="pUC ori"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          complement(2580..2599)
                     /label="pUCoriR"
                     /note="sequence: tatagtcctgtcgggtttcg"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     CDS             complement(3278..4138)
                     /label="Amp (bla)"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     CDS             complement(3278..4138)
                     /label="Translation 4138-3278"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW*"
     primer          complement(3403..3421)
                     /label="EKo152.2"
                     /note="sequence: tctcgcggtatcattgcag"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     promoter        complement(4139..4237)
                     /label="bla promoter"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          4233..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 agAGGACCTA CGCTGCCCTA GAGGTTTTGC TAGGGAGGAG ACGTGTGTGG CTGTAGCCAC
      601 CCGTCCCGGG TACAAGTCCC GGGTGGTGAG GACGGTGTCT GTGGTTGTCT TCCCAGACTC
      661 TGCTTTCTGC CGTCTTCGGT CAAGTACCAG CTGGTGGTCC GCATGTTTTg ggcccTTTTT
      721 Tggccgcagg aaccccctag aagttcctat tccgaagttc ctattctcta gaaagtatag
      781 gaacttcctt ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg
      841 aaaagttcga cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt
      901 tcagcttcga tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt
      961 tctacaaaga tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag
     1021 tgcttgacat tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg
     1081 gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg
     1141 aggccatgga tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg
     1201 gaccgcaagg aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc
     1261 cccatgtgta tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg
     1321 ctctcgatga gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg
     1381 cggatttcgg ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact
     1441 ggagcgaggc gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc
     1501 cgtggttggc ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg
     1561 caggatcgcc gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga
     1621 gcttggttga cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg
     1681 tccgatccgg agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct
     1741 ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc
     1801 cgagggcaaa ggaatagcac gtactacgag atttcgattc caccgccgcc ttctatgaaa
     1861 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1921 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     1981 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     2041 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     2101 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     2161 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     2221 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     2281 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     2341 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     2401 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     2461 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     2521 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     2581 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     2641 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2701 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2761 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2821 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2881 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2941 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     3001 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     3061 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     3121 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     3181 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     3241 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3301 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3361 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3421 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3481 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3541 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     3601 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3661 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3721 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3781 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3841 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3901 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3961 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4021 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4081 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4141 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     4201 tatttgaatg tatttagaaa aataaacaaa taggggt
//