LOCUS       01_pET-6xHis-GFP(-6)-L1 5914 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..864
                     /label="6xHis-GFP(-6)-L1"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /translation="MGHHHHHHGGASKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKKGRRGKKSRK*"
     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..861
                     /label="L1 Peptide"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(959..977)
                     /label="T7_Terminal_primer"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     gene            complement(1627..2487)
                     /label="Ampicillin"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     promoter        complement(2530..2558)
                     /label="AmpR_promoter"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     origin          2645..3264
                     /label="pBR322_origin"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          3663..3685
                     /label="pGEX_3_primer"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     feature         complement(3679..3870)
                     /label="ROP"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     regulatory      complement(4442..5533)
                     /label="lacI"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     gene            complement(5625..5876)
                     /label="tet (887 - 636)"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          5808..5827
                     /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 AACTCTACAA GAAAGGACGA
      841 AGAGGCAAGA AATCTCGAAA GTAACTCGAG TCTGGTAAAG AAACCGCTGC TGCGAAATTT
      901 GAACGCCAGC ACATGGACTC GTCTACTAGC GCAGCTTAAT TAACCTAGGC TGCTGCCACC
      961 GCTGAGCAAT AACTAGCATA ACCCCTTGGG GCCTCTAAAC GGGTCTTGAG GGGTTTTTTG
     1021 CTGAAAGGAG GAACTATATC CGGATTGGCG AATGGGACGC GCCCTGTAGC GGCGCATTAA
     1081 GCGCGGCGGG TGTGGTGGTT ACGCGCAGCG TGACCGCTAC ACTTGCCAGC GCCCTAGCGC
     1141 CCGCTCCTTT CGCTTTCTTC CCTTCCTTTC TCGCCACGTT CGCCGGCTTT CCCCGTCAAG
     1201 CTCTAAATCG GGGGCTCCCT TTAGGGTTCC GATTTAGTGC TTTACGGCAC CTCGACCCCA
     1261 AAAAACTTGA TTAGGGTGAT GGTTCACGTA GTGGGCCATC GCCCTGATAG ACGGTTTTTC
     1321 GCCCTTTGAC GTTGGAGTCC ACGTTCTTTA ATAGTGGACT CTTGTTCCAA ACTGGAACAA
     1381 CACTCAACCC TATCTCGGTC TATTCTTTTG ATTTATAAGG GATTTTGCCG ATTTCGGCCT
     1441 ATTGGTTAAA AAATGAGCTG ATTTAACAAA AATTTAACGC GAATTTTAAC AAAATATTAA
     1501 CGTTTACAAT TTCTGGCGGC ACGATGGCAT GAGATTATCA AAAAGGATCT TCACCTAGAT
     1561 CCTTTTAAAT TAAAAATGAA GTTTTAAATC AATCTAAAGT ATATATGAGT AAACTTGGTC
     1621 TGACAGTTAC CAATGCTTAA TCAGTGAGGC ACCTATCTCA GCGATCTGTC TATTTCGTTC
     1681 ATCCATAGTT GCCTGACTCC CCGTCGTGTA GATAACTACG ATACGGGAGG GCTTACCATC
     1741 TGGCCCCAGT GCTGCAATGA TACCGCGAGA CCCACGCTCA CCGGCTCCAG ATTTATCAGC
     1801 AATAAACCAG CCAGCCGGAA GGGCCGAGCG CAGAAGTGGT CCTGCAACTT TATCCGCCTC
     1861 CATCCAGTCT ATTAATTGTT GCCGGGAAGC TAGAGTAAGT AGTTCGCCAG TTAATAGTTT
     1921 GCGCAACGTT GTTGCCATTG CTACAGGCAT CGTGGTGTCA CGCTCGTCGT TTGGTATGGC
     1981 TTCATTCAGC TCCGGTTCCC AACGATCAAG GCGAGTTACA TGATCCCCCA TGTTGTGCAA
     2041 AAAAGCGGTT AGCTCCTTCG GTCCTCCGAT CGTTGTCAGA AGTAAGTTGG CCGCAGTGTT
     2101 ATCACTCATG GTTATGGCAG CACTGCATAA TTCTCTTACT GTCATGCCAT CCGTAAGATG
     2161 CTTTTCTGTG ACTGGTGAGT ACTCAACCAA GTCATTCTGA GAATAGTGTA TGCGGCGACC
     2221 GAGTTGCTCT TGCCCGGCGT CAATACGGGA TAATACCGCG CCACATAGCA GAACTTTAAA
     2281 AGTGCTCATC ATTGGAAAAC GTTCTTCGGG GCGAAAACTC TCAAGGATCT TACCGCTGTT
     2341 GAGATCCAGT TCGATGTAAC CCACTCGTGC ACCCAACTGA TCTTCAGCAT CTTTTACTTT
     2401 CACCAGCGTT TCTGGGTGAG CAAAAACAGG AAGGCAAAAT GCCGCAAAAA AGGGAATAAG
     2461 GGCGACACGG AAATGTTGAA TACTCATACT CTTCCTTTTT CAATCATGAT TGAAGCATTT
     2521 ATCAGGGTTA TTGTCTCATG AGCGGATACA TATTTGAATG TATTTAGAAA AATAAACAAA
     2581 TAGGTCATGA CCAAAATCCC TTAACGTGAG TTTTCGTTCC ACTGAGCGTC AGACCCCGTA
     2641 GAAAAGATCA AAGGATCTTC TTGAGATCCT TTTTTTCTGC GCGTAATCTG CTGCTTGCAA
     2701 ACAAAAAAAC CACCGCTACC AGCGGTGGTT TGTTTGCCGG ATCAAGAGCT ACCAACTCTT
     2761 TTTCCGAAGG TAACTGGCTT CAGCAGAGCG CAGATACCAA ATACTGTCCT TCTAGTGTAG
     2821 CCGTAGTTAG GCCACCACTT CAAGAACTCT GTAGCACCGC CTACATACCT CGCTCTGCTA
     2881 ATCCTGTTAC CAGTGGCTGC TGCCAGTGGC GATAAGTCGT GTCTTACCGG GTTGGACTCA
     2941 AGACGATAGT TACCGGATAA GGCGCAGCGG TCGGGCTGAA CGGGGGGTTC GTGCACACAG
     3001 CCCAGCTTGG AGCGAACGAC CTACACCGAA CTGAGATACC TACAGCGTGA GCTATGAGAA
     3061 AGCGCCACGC TTCCCGAAGG GAGAAAGGCG GACAGGTATC CGGTAAGCGG CAGGGTCGGA
     3121 ACAGGAGAGC GCACGAGGGA GCTTCCAGGG GGAAACGCCT GGTATCTTTA TAGTCCTGTC
     3181 GGGTTTCGCC ACCTCTGACT TGAGCGTCGA TTTTTGTGAT GCTCGTCAGG GGGGCGGAGC
     3241 CTATGGAAAA ACGCCAGCAA CGCGGCCTTT TTACGGTTCC TGGCCTTTTG CTGGCCTTTT
     3301 GCTCACATGT TCTTTCCTGC GTTATCCCCT GATTCTGTGG ATAACCGTAT TACCGCCTTT
     3361 GAGTGAGCTG ATACCGCTCG CCGCAGCCGA ACGACCGAGC GCAGCGAGTC AGTGAGCGAG
     3421 GAAGCGGAAG AGCGCCTGAT GCGGTATTTT CTCCTTACGC ATCTGTGCGG TATTTCACAC
     3481 CGCATATATG GTGCACTCTC AGTACAATCT GCTCTGATGC CGCATAGTTA AGCCAGTATA
     3541 CACTCCGCTA TCGCTACGTG ACTGGGTCAT GGCTGCGCCC CGACACCCGC CAACACCCGC
     3601 TGACGCGCCC TGACGGGCTT GTCTGCTCCC GGCATCCGCT TACAGACAAG CTGTGACCGT
     3661 CTCCGGGAGC TGCATGTGTC AGAGGTTTTC ACCGTCATCA CCGAAACGCG CGAGGCAGCT
     3721 GCGGTAAAGC TCATCAGCGT GGTCGTGAAG CGATTCACAG ATGTCTGCCT GTTCATCCGC
     3781 GTCCAGCTCG TTGAGTTTCT CCAGAAGCGT TAATGTCTGG CTTCTGATAA AGCGGGCCAT
     3841 GTTAAGGGCG GTTTTTTCCT GTTTGGTCAC TGATGCCTCC GTGTAAGGGG GATTTCTGTT
     3901 CATGGGGGTA ATGATACCGA TGAAACGAGA GAGGATGCTC ACGATACGGG TTACTGATGA
     3961 TGAACATGCC CGGTTACTGG AACGTTGTGA GGGTAAACAA CTGGCGGTAT GGATGCGGCG
     4021 GGACCAGAGA AAAATCACTC AGGGTCAATG CCAGCGCTTC GTTAATACAG ATGTAGGTGT
     4081 TCCACAGGGT AGCCAGCAGC ATCCTGCGAT GCAGATCCGG AACATAATGG TGCAGGGCGC
     4141 TGACTTCCGC GTTTCCAGAC TTTACGAAAC ACGGAAACCG AAGACCATTC ATGTTGTTGC
     4201 TCAGGTCGCA GACGTTTTGC AGCAGCAGTC GCTTCACGTT CGCTCGCGTA TCGGTGATTC
     4261 ATTCTGCTAA CCAGTAAGGC AACCCCGCCA GCCTAGCCGG GTCCTCAACG ACAGGAGCAC
     4321 GATCATGCTA GTCATGCCCC GCGCCCACCG GAAGGAGCTG ACTGGGTTGA AGGCTCTCAA
     4381 GGGCATCGGT CGAGATCCCG GTGCCTAATG AGTGAGCTAA CTTACATTAA TTGCGTTGCG
     4441 CTCACTGCCC GCTTTCCAGT CGGGAAACCT GTCGTGCCAG CTGCATTAAT GAATCGGCCA
     4501 ACGCGCGGGG AGAGGCGGTT TGCGTATTGG GCGCCAGGGT GGTTTTTCTT TTCACCAGTG
     4561 AGACGGGCAA CAGCTGATTG CCCTTCACCG CCTGGCCCTG AGAGAGTTGC AGCAAGCGGT
     4621 CCACGCTGGT TTGCCCCAGC AGGCGAAAAT CCTGTTTGAT GGTGGTTAAC GGCGGGATAT
     4681 AACATGAGCT GTCTTCGGTA TCGTCGTATC CCACTACCGA GATGTCCGCA CCAACGCGCA
     4741 GCCCGGACTC GGTAATGGCG CGCATTGCGC CCAGCGCCAT CTGATCGTTG GCAACCAGCA
     4801 TCGCAGTGGG AACGATGCCC TCATTCAGCA TTTGCATGGT TTGTTGAAAA CCGGACATGG
     4861 CACTCCAGTC GCCTTCCCGT TCCGCTATCG GCTGAATTTG ATTGCGAGTG AGATATTTAT
     4921 GCCAGCCAGC CAGACGCAGA CGCGCCGAGA CAGAACTTAA TGGGCCCGCT AACAGCGCGA
     4981 TTTGCTGGTG ACCCAATGCG ACCAGATGCT CCACGCCCAG TCGCGTACCG TCTTCATGGG
     5041 AGAAAATAAT ACTGTTGATG GGTGTCTGGT CAGAGACATC AAGAAATAAC GCCGGAACAT
     5101 TAGTGCAGGC AGCTTCCACA GCAATGGCAT CCTGGTCATC CAGCGGATAG TTAATGATCA
     5161 GCCCACTGAC GCGTTGCGCG AGAAGATTGT GCACCGCCGC TTTACAGGCT TCGACGCCGC
     5221 TTCGTTCTAC CATCGACACC ACCACGCTGG CACCCAGTTG ATCGGCGCGA GATTTAATCG
     5281 CCGCGACAAT TTGCGACGGC GCGTGCAGGG CCAGACTGGA GGTGGCAACG CCAATCAGCA
     5341 ACGACTGTTT GCCCGCCAGT TGTTGTGCCA CGCGGTTGGG AATGTAATTC AGCTCCGCCA
     5401 TCGCCGCTTC CACTTTTTCC CGCGTTTTCG CAGAAACGTG GCTGGCCTGG TTCACCACGC
     5461 GGGAAACGGT CTGATAAGAG ACACCGGCAT ACTCTGCGAC ATCGTATAAC GTTACTGGTT
     5521 TCACATTCAC CACCCTGAAT TGACTCTCTT CCGGGCGCTA TCATGCCATA CCGCGAAAGG
     5581 TTTTGCGCCA TTCGATGGTG TCCGGGATCT CGACGCTCTC CCTTATGCGA CTCCTGCATT
     5641 AGGAAGCAGC CCAGTAGTAG GTTGAGGCCG TTGAGCACCG CCGCCGCAAG GAATGGTGCA
     5701 TGCAAGGAGA TGGCGCCCAA CAGTCCCCCG GCCACGGGGC CTGCCACCAT ACCCACGCCG
     5761 AAACAAGCGC TCATGAGCCC GAAGTGGCGA GCCCGATCTT CCCCATCGGT GATGTCGGCG
     5821 ATATAGGCGC CAGCAACCGC ACCTGTGGCG CCGGTGATGC CGGCCACGAT GCGTCCGGCG
     5881 TAGAGGATCG AGATCGATCT CGATCCCGCG AAAT
//