LOCUS       06_pET-6xHis-GFP(-6)-H3 5944 bp ds-DNA     circular     29-JUN-2023
DEFINITION  .
FEATURES             Location/Qualifiers
     promoter        1..17
                     /label="T7_promoter"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     promoter        18..19
                     /label="T7_promoter"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     regulatory      19..46
                     /label="lacO"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     feature         64..80
                     /label="T7_transl_en_RBS"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             88..894
                     /label="6xHis-GFP(-6)-H3"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /translation="MGHHHHHHGGASKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKKKRRRKKKRRRKKKRRRKK**"
     feature         94..111
                     /label="6xHIS"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     mutation        133..135
                     /label="E16R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        142..144
                     /label="T19R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        148..150
                     /label="V21K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        172..174
                     /label="D29K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        211..213
                     /label="E42K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        217..219
                     /label="E44K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        232..234
                     /label="N49R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     feature         298..327
                     /label="Y66 (EGFP)"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     mutation        343..345
                     /label="D86K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        355..357
                     /label="Q90R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        385..387
                     /label="E100K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        421..423
                     /label="D112K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        430..432
                     /label="T115K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        466..468
                     /label="D127R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        487..489
                     /label="E134K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        499..501
                     /label="I138R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        514..516
                     /label="D143K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        541..543
                     /label="E152R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        562..564
                     /label="N159K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        586..588
                     /label="Q167R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        607..609
                     /label="N174K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        631..633
                     /label="E182K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        685..687
                     /label="D200R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        706..708
                     /label="D207R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        727..729
                     /label="Q214R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        733..735
                     /label="A216K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        751..753
                     /label="N222K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        805..807
                     /label="T240K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        814..816
                     /label="M243R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        823..825
                     /label="L246R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    835..888
                     /label="H3 Peptide"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(989..1007)
                     /label="T7_Terminal_primer"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     gene            complement(1657..2517)
                     /label="Ampicillin"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     promoter        complement(2560..2588)
                     /label="AmpR_promoter"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     origin          2675..3294
                     /label="pBR322_origin"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          3693..3715
                     /label="pGEX_3_primer"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     feature         complement(3709..3900)
                     /label="ROP"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     regulatory      complement(4472..5563)
                     /label="lacI"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     gene            complement(5655..5906)
                     /label="tet (887 - 636)"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          5838..5857
                     /label="pBRrevBam_primer"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
ORIGIN
        1 TAATACGACT CACTATAGGG GAATTGTGAG CGGATAACAA TTCCCCTCTA GAAATAATTT
       61 TGTTTAACTT TAAGAAGGAG ATATACCATG GGTCATCACC ACCACCATCA CGGTGGCGCT
      121 AGCAAAGGTG AAGAGCTGTT TACGGGTGTA GTACCGATCT TAGTGGAATT AGACGGCGAC
      181 GTGAACGGTC ACAAATTTAG CGTGCGCGGC GAAGGCGAAG GTGACGCTAC CAATGGTAAA
      241 TTGACCCTGA AGTTTATTTG CACAACAGGC AAATTACCCG TTCCGTGGCC CACCTTAGTG
      301 ACCACCCTGA CCTATGGCGT TCAGTGCTTC AGTCGTTACC CAGATCATAT GAAACAACAC
      361 GATTTTTTCA AATCAGCCAT GCCTGAAGGA TATGTTCAAG AGCGTACAAT CAGCTTCAAG
      421 GACGATGGCA CCTATAAAAC GCGTGCGGAA GTGAAATTTG AAGGCGACAC ATTAGTAAAC
      481 CGTATCGAAC TGAAAGGTAT CGACTTCAAA GAAGACGGCA ACATTTTAGG CCATAAGCTG
      541 GAATATAACT TTAATTCTCA TAACGTGTAT ATTACGGCCG ATAAACAGAA AAACGGTATC
      601 AAGGCAAATT TCAAAATTCG CCATAACGTG GAAGACGGCA GCGTTCAATT AGCGGATCAT
      661 TATCAACAAA ACACGCCGAT TGGTGACGGG CCTGTACTGT TACCTGACAA CCACTACCTG
      721 AGCACCCAGT CAGCACTGAG CAAAGATCCG AACGAAAAAC GCGATCACAT GGTTCTGTTA
      781 GAATTCGTGA CCGCTGCAGG CATTACTCAC GGAATGGACG AGCTGTACAA GAAAAAGAGG
      841 AGAAGGAAGA AAAAGCGTAG ACGCAAGAAA AAGCGTAGAC GCAAGAAATA ATAACTCGAG
      901 TCTGGTAAAG AAACCGCTGC TGCGAAATTT GAACGCCAGC ACATGGACTC GTCTACTAGC
      961 GCAGCTTAAT TAACCTAGGC TGCTGCCACC GCTGAGCAAT AACTAGCATA ACCCCTTGGG
     1021 GCCTCTAAAC GGGTCTTGAG GGGTTTTTTG CTGAAAGGAG GAACTATATC CGGATTGGCG
     1081 AATGGGACGC GCCCTGTAGC GGCGCATTAA GCGCGGCGGG TGTGGTGGTT ACGCGCAGCG
     1141 TGACCGCTAC ACTTGCCAGC GCCCTAGCGC CCGCTCCTTT CGCTTTCTTC CCTTCCTTTC
     1201 TCGCCACGTT CGCCGGCTTT CCCCGTCAAG CTCTAAATCG GGGGCTCCCT TTAGGGTTCC
     1261 GATTTAGTGC TTTACGGCAC CTCGACCCCA AAAAACTTGA TTAGGGTGAT GGTTCACGTA
     1321 GTGGGCCATC GCCCTGATAG ACGGTTTTTC GCCCTTTGAC GTTGGAGTCC ACGTTCTTTA
     1381 ATAGTGGACT CTTGTTCCAA ACTGGAACAA CACTCAACCC TATCTCGGTC TATTCTTTTG
     1441 ATTTATAAGG GATTTTGCCG ATTTCGGCCT ATTGGTTAAA AAATGAGCTG ATTTAACAAA
     1501 AATTTAACGC GAATTTTAAC AAAATATTAA CGTTTACAAT TTCTGGCGGC ACGATGGCAT
     1561 GAGATTATCA AAAAGGATCT TCACCTAGAT CCTTTTAAAT TAAAAATGAA GTTTTAAATC
     1621 AATCTAAAGT ATATATGAGT AAACTTGGTC TGACAGTTAC CAATGCTTAA TCAGTGAGGC
     1681 ACCTATCTCA GCGATCTGTC TATTTCGTTC ATCCATAGTT GCCTGACTCC CCGTCGTGTA
     1741 GATAACTACG ATACGGGAGG GCTTACCATC TGGCCCCAGT GCTGCAATGA TACCGCGAGA
     1801 CCCACGCTCA CCGGCTCCAG ATTTATCAGC AATAAACCAG CCAGCCGGAA GGGCCGAGCG
     1861 CAGAAGTGGT CCTGCAACTT TATCCGCCTC CATCCAGTCT ATTAATTGTT GCCGGGAAGC
     1921 TAGAGTAAGT AGTTCGCCAG TTAATAGTTT GCGCAACGTT GTTGCCATTG CTACAGGCAT
     1981 CGTGGTGTCA CGCTCGTCGT TTGGTATGGC TTCATTCAGC TCCGGTTCCC AACGATCAAG
     2041 GCGAGTTACA TGATCCCCCA TGTTGTGCAA AAAAGCGGTT AGCTCCTTCG GTCCTCCGAT
     2101 CGTTGTCAGA AGTAAGTTGG CCGCAGTGTT ATCACTCATG GTTATGGCAG CACTGCATAA
     2161 TTCTCTTACT GTCATGCCAT CCGTAAGATG CTTTTCTGTG ACTGGTGAGT ACTCAACCAA
     2221 GTCATTCTGA GAATAGTGTA TGCGGCGACC GAGTTGCTCT TGCCCGGCGT CAATACGGGA
     2281 TAATACCGCG CCACATAGCA GAACTTTAAA AGTGCTCATC ATTGGAAAAC GTTCTTCGGG
     2341 GCGAAAACTC TCAAGGATCT TACCGCTGTT GAGATCCAGT TCGATGTAAC CCACTCGTGC
     2401 ACCCAACTGA TCTTCAGCAT CTTTTACTTT CACCAGCGTT TCTGGGTGAG CAAAAACAGG
     2461 AAGGCAAAAT GCCGCAAAAA AGGGAATAAG GGCGACACGG AAATGTTGAA TACTCATACT
     2521 CTTCCTTTTT CAATCATGAT TGAAGCATTT ATCAGGGTTA TTGTCTCATG AGCGGATACA
     2581 TATTTGAATG TATTTAGAAA AATAAACAAA TAGGTCATGA CCAAAATCCC TTAACGTGAG
     2641 TTTTCGTTCC ACTGAGCGTC AGACCCCGTA GAAAAGATCA AAGGATCTTC TTGAGATCCT
     2701 TTTTTTCTGC GCGTAATCTG CTGCTTGCAA ACAAAAAAAC CACCGCTACC AGCGGTGGTT
     2761 TGTTTGCCGG ATCAAGAGCT ACCAACTCTT TTTCCGAAGG TAACTGGCTT CAGCAGAGCG
     2821 CAGATACCAA ATACTGTCCT TCTAGTGTAG CCGTAGTTAG GCCACCACTT CAAGAACTCT
     2881 GTAGCACCGC CTACATACCT CGCTCTGCTA ATCCTGTTAC CAGTGGCTGC TGCCAGTGGC
     2941 GATAAGTCGT GTCTTACCGG GTTGGACTCA AGACGATAGT TACCGGATAA GGCGCAGCGG
     3001 TCGGGCTGAA CGGGGGGTTC GTGCACACAG CCCAGCTTGG AGCGAACGAC CTACACCGAA
     3061 CTGAGATACC TACAGCGTGA GCTATGAGAA AGCGCCACGC TTCCCGAAGG GAGAAAGGCG
     3121 GACAGGTATC CGGTAAGCGG CAGGGTCGGA ACAGGAGAGC GCACGAGGGA GCTTCCAGGG
     3181 GGAAACGCCT GGTATCTTTA TAGTCCTGTC GGGTTTCGCC ACCTCTGACT TGAGCGTCGA
     3241 TTTTTGTGAT GCTCGTCAGG GGGGCGGAGC CTATGGAAAA ACGCCAGCAA CGCGGCCTTT
     3301 TTACGGTTCC TGGCCTTTTG CTGGCCTTTT GCTCACATGT TCTTTCCTGC GTTATCCCCT
     3361 GATTCTGTGG ATAACCGTAT TACCGCCTTT GAGTGAGCTG ATACCGCTCG CCGCAGCCGA
     3421 ACGACCGAGC GCAGCGAGTC AGTGAGCGAG GAAGCGGAAG AGCGCCTGAT GCGGTATTTT
     3481 CTCCTTACGC ATCTGTGCGG TATTTCACAC CGCATATATG GTGCACTCTC AGTACAATCT
     3541 GCTCTGATGC CGCATAGTTA AGCCAGTATA CACTCCGCTA TCGCTACGTG ACTGGGTCAT
     3601 GGCTGCGCCC CGACACCCGC CAACACCCGC TGACGCGCCC TGACGGGCTT GTCTGCTCCC
     3661 GGCATCCGCT TACAGACAAG CTGTGACCGT CTCCGGGAGC TGCATGTGTC AGAGGTTTTC
     3721 ACCGTCATCA CCGAAACGCG CGAGGCAGCT GCGGTAAAGC TCATCAGCGT GGTCGTGAAG
     3781 CGATTCACAG ATGTCTGCCT GTTCATCCGC GTCCAGCTCG TTGAGTTTCT CCAGAAGCGT
     3841 TAATGTCTGG CTTCTGATAA AGCGGGCCAT GTTAAGGGCG GTTTTTTCCT GTTTGGTCAC
     3901 TGATGCCTCC GTGTAAGGGG GATTTCTGTT CATGGGGGTA ATGATACCGA TGAAACGAGA
     3961 GAGGATGCTC ACGATACGGG TTACTGATGA TGAACATGCC CGGTTACTGG AACGTTGTGA
     4021 GGGTAAACAA CTGGCGGTAT GGATGCGGCG GGACCAGAGA AAAATCACTC AGGGTCAATG
     4081 CCAGCGCTTC GTTAATACAG ATGTAGGTGT TCCACAGGGT AGCCAGCAGC ATCCTGCGAT
     4141 GCAGATCCGG AACATAATGG TGCAGGGCGC TGACTTCCGC GTTTCCAGAC TTTACGAAAC
     4201 ACGGAAACCG AAGACCATTC ATGTTGTTGC TCAGGTCGCA GACGTTTTGC AGCAGCAGTC
     4261 GCTTCACGTT CGCTCGCGTA TCGGTGATTC ATTCTGCTAA CCAGTAAGGC AACCCCGCCA
     4321 GCCTAGCCGG GTCCTCAACG ACAGGAGCAC GATCATGCTA GTCATGCCCC GCGCCCACCG
     4381 GAAGGAGCTG ACTGGGTTGA AGGCTCTCAA GGGCATCGGT CGAGATCCCG GTGCCTAATG
     4441 AGTGAGCTAA CTTACATTAA TTGCGTTGCG CTCACTGCCC GCTTTCCAGT CGGGAAACCT
     4501 GTCGTGCCAG CTGCATTAAT GAATCGGCCA ACGCGCGGGG AGAGGCGGTT TGCGTATTGG
     4561 GCGCCAGGGT GGTTTTTCTT TTCACCAGTG AGACGGGCAA CAGCTGATTG CCCTTCACCG
     4621 CCTGGCCCTG AGAGAGTTGC AGCAAGCGGT CCACGCTGGT TTGCCCCAGC AGGCGAAAAT
     4681 CCTGTTTGAT GGTGGTTAAC GGCGGGATAT AACATGAGCT GTCTTCGGTA TCGTCGTATC
     4741 CCACTACCGA GATGTCCGCA CCAACGCGCA GCCCGGACTC GGTAATGGCG CGCATTGCGC
     4801 CCAGCGCCAT CTGATCGTTG GCAACCAGCA TCGCAGTGGG AACGATGCCC TCATTCAGCA
     4861 TTTGCATGGT TTGTTGAAAA CCGGACATGG CACTCCAGTC GCCTTCCCGT TCCGCTATCG
     4921 GCTGAATTTG ATTGCGAGTG AGATATTTAT GCCAGCCAGC CAGACGCAGA CGCGCCGAGA
     4981 CAGAACTTAA TGGGCCCGCT AACAGCGCGA TTTGCTGGTG ACCCAATGCG ACCAGATGCT
     5041 CCACGCCCAG TCGCGTACCG TCTTCATGGG AGAAAATAAT ACTGTTGATG GGTGTCTGGT
     5101 CAGAGACATC AAGAAATAAC GCCGGAACAT TAGTGCAGGC AGCTTCCACA GCAATGGCAT
     5161 CCTGGTCATC CAGCGGATAG TTAATGATCA GCCCACTGAC GCGTTGCGCG AGAAGATTGT
     5221 GCACCGCCGC TTTACAGGCT TCGACGCCGC TTCGTTCTAC CATCGACACC ACCACGCTGG
     5281 CACCCAGTTG ATCGGCGCGA GATTTAATCG CCGCGACAAT TTGCGACGGC GCGTGCAGGG
     5341 CCAGACTGGA GGTGGCAACG CCAATCAGCA ACGACTGTTT GCCCGCCAGT TGTTGTGCCA
     5401 CGCGGTTGGG AATGTAATTC AGCTCCGCCA TCGCCGCTTC CACTTTTTCC CGCGTTTTCG
     5461 CAGAAACGTG GCTGGCCTGG TTCACCACGC GGGAAACGGT CTGATAAGAG ACACCGGCAT
     5521 ACTCTGCGAC ATCGTATAAC GTTACTGGTT TCACATTCAC CACCCTGAAT TGACTCTCTT
     5581 CCGGGCGCTA TCATGCCATA CCGCGAAAGG TTTTGCGCCA TTCGATGGTG TCCGGGATCT
     5641 CGACGCTCTC CCTTATGCGA CTCCTGCATT AGGAAGCAGC CCAGTAGTAG GTTGAGGCCG
     5701 TTGAGCACCG CCGCCGCAAG GAATGGTGCA TGCAAGGAGA TGGCGCCCAA CAGTCCCCCG
     5761 GCCACGGGGC CTGCCACCAT ACCCACGCCG AAACAAGCGC TCATGAGCCC GAAGTGGCGA
     5821 GCCCGATCTT CCCCATCGGT GATGTCGGCG ATATAGGCGC CAGCAACCGC ACCTGTGGCG
     5881 CCGGTGATGC CGGCCACGAT GCGTCCGGCG TAGAGGATCG AGATCGATCT CGATCCCGCG
     5941 AAAT
//