LOCUS       CMV-TO-nTEVP-rbdK65E_(F 5801 bp ds-DNA     circular     11-JUN-2021
DEFINITION  .
FEATURES             Location/Qualifiers
     CDS             complement(585..1445)
                     /label="Amp (bla)"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     promoter        complement(1446..1544)
                     /label="bla promoter"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     promoter        1808..2435
                     /label="CMV"
                     /ApEinfo_revcolor=#c6c9d1
                     /ApEinfo_fwdcolor=#c6c9d1
     enhancer        1808..2534
                     /label="enhancer"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     TATA_signal     2380..2386
                     /label="TATA box"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    2396..2414
                     /label="TetO"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    2396..2435
                     /label="TetR binding sites"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    2417..2435
                     /label="TetO"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    2559..2566
                     /label="Kozak"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     CDS             2567..3196
                     /label="Translation 2567-3196"
     CDS             2576..2929
                     /label="N-TEV"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     CDS             2948..3193
                     /label="CRaf rbd K65E K108A"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     misc_feature    2993..2995
                     /label="K65E"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     misc_feature    3122..3124
                     /label="K108A"
                     /ApEinfo_revcolor=#f58a5e
                     /ApEinfo_fwdcolor=#f58a5e
     CDS             3211..3243
                     /label="c-myc epitope"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     misc_feature    3243..3245
                     /label="New Stop"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     CDS             3256..3273
                     /label="Polyhistidine tag"
                     /ApEinfo_revcolor=#993366
                     /ApEinfo_fwdcolor=#993366
     misc_feature    3274..3276
                     /label="STOP (missing now)"
                     /ApEinfo_revcolor=#a187be
                     /ApEinfo_fwdcolor=#a187be
     polyA_signal    3295..3509
                     /label="BGH polyadenylation sequence"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     polyA_signal    3542..3565
                     /label="BGH pA"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     misc_feature    3848..3895
                     /label="FRT"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     gene            3903..4923
                     /label="Hygromycin"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     polyA_signal    5055..5185
                     /label="SV40 pA"
                     /ApEinfo_revcolor=#b4abac
                     /ApEinfo_fwdcolor=#b4abac
     rep_origin      join(5568..5801,1..440)
                     /label="pUC ori"
                     /ApEinfo_revcolor=#ffef86
                     /ApEinfo_fwdcolor=#ffef86
ORIGIN
        1 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
       61 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
      121 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
      181 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
      241 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
      301 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
      361 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
      421 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
      481 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
      541 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
      601 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
      661 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
      721 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
      781 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
      841 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
      901 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
      961 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     1021 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     1081 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     1141 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     1201 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     1261 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     1321 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     1381 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     1441 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     1501 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     1561 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     1621 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     1681 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     1741 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgt acgggccaga
     1801 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     1861 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     1921 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     1981 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     2041 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     2101 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     2161 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2221 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2281 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2341 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct
     2401 atcagtgata gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac
     2461 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
     2521 cgatccagcc tccggactct agcgtttaaa cttaagcttc gccaccatgg gatccGGGGA
     2581 GAGTTTGTTT AAGGGGCCAA GGGACTATAA CCCAATATCA TCCACTATTT GCCACCTCAC
     2641 TAACGAGAGC GATGGACATA CAACCTCTCT CTACGGGATA GGCTTCGGTC CTTTCATCAT
     2701 TACCAATAAG CATCTGTTTC GCCGAAACAA CGGTACTCTT CTGGTTCAAT CTCTTCATGG
     2761 TGTCTTCAAG GTGAAAAACA CCACTACGCT TCAGCAACAC CTGATTGATG GTAGGGATAT
     2821 GATAATTATC AGAATGCCGA AAGATTTCCC ACCTTTTCCA CAGAAGCTGA AATTCAGGGA
     2881 ACCGCAGAGA GAGGAGAGGA TTTGTTTGGT AACGACCAAC TTCCAGACGG GCTCTGGCAG
     2941 CAGCGCGGAT CCGAGTAAGA CGTCAAACAC AATCAGGGTT TTTCTCCCCA ATgAACAGCG
     3001 GACCGTGGTA AACGTCCGAA ACGGGATGAG TCTCCACGAC TGTCTGATGA AAGCCTTGAA
     3061 GGTGAGGGGG CTTCAGCCTG AATGTTGCGC TGTCTTTCGC CTTCTCCACG AACACAAGGG
     3121 CGCAAAGGCG AGGCTTGACT GGAATACAGA CGCTGCAAGT TTGATTGGGG AAGAACTTCA
     3181 GGTCGATTTC CTCtagctcg aggagggccc gaacaaaaac tcatctcaga agaggatctg
     3241 aatagcgccg tcgaccatca tcatcatcat cattgagttt aaacccgctg atcagcctcg
     3301 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     3361 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     3421 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     3481 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggagat ctatcgatgt
     3541 tctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     3601 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3661 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     3721 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggtc cctttagggt
     3781 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3841 gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga acttccttgg
     3901 ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca
     3961 gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg
     4021 taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc
     4081 gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg
     4141 gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc
     4201 aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg
     4261 cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa
     4321 tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc
     4381 actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc
     4441 tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct
     4501 ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga
     4561 tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt
     4621 gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc
     4681 ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg
     4741 gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag
     4801 ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct
     4861 gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg
     4921 aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4981 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     5041 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5101 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5161 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     5221 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5281 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     5341 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5401 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5461 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5521 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5581 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5641 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5701 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5761 ccctgccgct taccggatac ctgtccgcct ttctcccttc g
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