LOCUS       15_pET-6xHis-GFP(+6)-L3 5968 bp ds-DNA     circular     26-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..918
                     /label="6xHis-GFP(+6)-L3"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /translation="MGHHHHHHGGASKGERLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPKHMKQHDFFKSAMPEGYVQERTISFKKDGTYKTRAEVKFEGRTLVNRIELKGRDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPRNHYLSTQSALSKDPKEKRDHMVLLEFVTAAGITHGMDERYKGRRGKKSRKGRRGKKSRKGRRGKKSRK**"
     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    832..912
                     /label="L3 Peptide"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(1013..1031)
                     /label="T7_Terminal_primer"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     gene            complement(1681..2541)
                     /label="Ampicillin"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     promoter        complement(2584..2612)
                     /label="AmpR_promoter"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     origin          2699..3318
                     /label="pBR322_origin"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          3717..3739
                     /label="pGEX_3_primer"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     feature         complement(3733..3924)
                     /label="ROP"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     regulatory      complement(4496..5587)
                     /label="lacI"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     gene            complement(5679..5930)
                     /label="tet (887 - 636)"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          5862..5881
                     /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 AACGTCTGTT TACTGGTGTA GTACCGATCT TAGTGGAATT AGACGGCGAC
      181 GTGAACGGTC ATAAATTTAG CGTGCGCGGC GAGGGCGAAG GTGACGCTAC CAATGGTAAA
      241 TTGACCCTGA AGTTTATTTG CACAACAGGC AAATTACCCG TTCCGTGGCC CACCTTAGTG
      301 ACCACCCTGA CCTATGGCGT TCAGTGCTTC AGTCGTTACC CTAAACATAT GAAACAACAC
      361 GATTTTTTCA AATCAGCCAT GCCTGAAGGA TATGTTCAAG AGCGTACAAT CAGCTTCAAG
      421 AAGGATGGCA CCTATAAAAC GCGTGCGGAA GTGAAATTTG AAGGCCGCAC ATTAGTAAAT
      481 CGTATCGAAC TGAAAGGTCG TGACTTCAAA GAAGACGGCA ACATTTTAGG CCATAAACTG
      541 GAATATAACT TTAATTCTCA TAACGTGTAT ATTACGGCCG ATAAACAGAA GAATGGTATC
      601 AAGGCAAATT TCAAAATTCG CCATAACGTG GAGGACGGCA GCGTTCAATT AGCGGATCAT
      661 TATCAACAAA ACACGCCGAT TGGTGATGGG CCTGTACTGT TACCTCGCAA CCACTACCTG
      721 AGCACCCAAT CTGCCCTGAG CAAAGATCCG AAAGAAAAAC GCGATCACAT GGTTCTGTTA
      781 GAATTCGTGA CCGCTGCAGG CATTACGCAC GGAATGGACG AACGCTACAA GGGTCGAAGA
      841 GGGAAGAAAT CGCGGAAGGG TCGAAGAGGG AAGAAATCGC GGAAGGGAAG AAGGGGTAAG
      901 AAATCTAGGA AGTAATAACT CGAGTCTGGT AAAGAAACCG CTGCTGCGAA ATTTGAACGC
      961 CAGCACATGG ACTCGTCTAC TAGCGCAGCT TAATTAACCT AGGCTGCTGC CACCGCTGAG
     1021 CAATAACTAG CATAACCCCT TGGGGCCTCT AAACGGGTCT TGAGGGGTTT TTTGCTGAAA
     1081 GGAGGAACTA TATCCGGATT GGCGAATGGG ACGCGCCCTG TAGCGGCGCA TTAAGCGCGG
     1141 CGGGTGTGGT GGTTACGCGC AGCGTGACCG CTACACTTGC CAGCGCCCTA GCGCCCGCTC
     1201 CTTTCGCTTT CTTCCCTTCC TTTCTCGCCA CGTTCGCCGG CTTTCCCCGT CAAGCTCTAA
     1261 ATCGGGGGCT CCCTTTAGGG TTCCGATTTA GTGCTTTACG GCACCTCGAC CCCAAAAAAC
     1321 TTGATTAGGG TGATGGTTCA CGTAGTGGGC CATCGCCCTG ATAGACGGTT TTTCGCCCTT
     1381 TGACGTTGGA GTCCACGTTC TTTAATAGTG GACTCTTGTT CCAAACTGGA ACAACACTCA
     1441 ACCCTATCTC GGTCTATTCT TTTGATTTAT AAGGGATTTT GCCGATTTCG GCCTATTGGT
     1501 TAAAAAATGA GCTGATTTAA CAAAAATTTA ACGCGAATTT TAACAAAATA TTAACGTTTA
     1561 CAATTTCTGG CGGCACGATG GCATGAGATT ATCAAAAAGG ATCTTCACCT AGATCCTTTT
     1621 AAATTAAAAA TGAAGTTTTA AATCAATCTA AAGTATATAT GAGTAAACTT GGTCTGACAG
     1681 TTACCAATGC TTAATCAGTG AGGCACCTAT CTCAGCGATC TGTCTATTTC GTTCATCCAT
     1741 AGTTGCCTGA CTCCCCGTCG TGTAGATAAC TACGATACGG GAGGGCTTAC CATCTGGCCC
     1801 CAGTGCTGCA ATGATACCGC GAGACCCACG CTCACCGGCT CCAGATTTAT CAGCAATAAA
     1861 CCAGCCAGCC GGAAGGGCCG AGCGCAGAAG TGGTCCTGCA ACTTTATCCG CCTCCATCCA
     1921 GTCTATTAAT TGTTGCCGGG AAGCTAGAGT AAGTAGTTCG CCAGTTAATA GTTTGCGCAA
     1981 CGTTGTTGCC ATTGCTACAG GCATCGTGGT GTCACGCTCG TCGTTTGGTA TGGCTTCATT
     2041 CAGCTCCGGT TCCCAACGAT CAAGGCGAGT TACATGATCC CCCATGTTGT GCAAAAAAGC
     2101 GGTTAGCTCC TTCGGTCCTC CGATCGTTGT CAGAAGTAAG TTGGCCGCAG TGTTATCACT
     2161 CATGGTTATG GCAGCACTGC ATAATTCTCT TACTGTCATG CCATCCGTAA GATGCTTTTC
     2221 TGTGACTGGT GAGTACTCAA CCAAGTCATT CTGAGAATAG TGTATGCGGC GACCGAGTTG
     2281 CTCTTGCCCG GCGTCAATAC GGGATAATAC CGCGCCACAT AGCAGAACTT TAAAAGTGCT
     2341 CATCATTGGA AAACGTTCTT CGGGGCGAAA ACTCTCAAGG ATCTTACCGC TGTTGAGATC
     2401 CAGTTCGATG TAACCCACTC GTGCACCCAA CTGATCTTCA GCATCTTTTA CTTTCACCAG
     2461 CGTTTCTGGG TGAGCAAAAA CAGGAAGGCA AAATGCCGCA AAAAAGGGAA TAAGGGCGAC
     2521 ACGGAAATGT TGAATACTCA TACTCTTCCT TTTTCAATCA TGATTGAAGC ATTTATCAGG
     2581 GTTATTGTCT CATGAGCGGA TACATATTTG AATGTATTTA GAAAAATAAA CAAATAGGTC
     2641 ATGACCAAAA TCCCTTAACG TGAGTTTTCG TTCCACTGAG CGTCAGACCC CGTAGAAAAG
     2701 ATCAAAGGAT CTTCTTGAGA TCCTTTTTTT CTGCGCGTAA TCTGCTGCTT GCAAACAAAA
     2761 AAACCACCGC TACCAGCGGT GGTTTGTTTG CCGGATCAAG AGCTACCAAC TCTTTTTCCG
     2821 AAGGTAACTG GCTTCAGCAG AGCGCAGATA CCAAATACTG TCCTTCTAGT GTAGCCGTAG
     2881 TTAGGCCACC ACTTCAAGAA CTCTGTAGCA CCGCCTACAT ACCTCGCTCT GCTAATCCTG
     2941 TTACCAGTGG CTGCTGCCAG TGGCGATAAG TCGTGTCTTA CCGGGTTGGA CTCAAGACGA
     3001 TAGTTACCGG ATAAGGCGCA GCGGTCGGGC TGAACGGGGG GTTCGTGCAC ACAGCCCAGC
     3061 TTGGAGCGAA CGACCTACAC CGAACTGAGA TACCTACAGC GTGAGCTATG AGAAAGCGCC
     3121 ACGCTTCCCG AAGGGAGAAA GGCGGACAGG TATCCGGTAA GCGGCAGGGT CGGAACAGGA
     3181 GAGCGCACGA GGGAGCTTCC AGGGGGAAAC GCCTGGTATC TTTATAGTCC TGTCGGGTTT
     3241 CGCCACCTCT GACTTGAGCG TCGATTTTTG TGATGCTCGT CAGGGGGGCG GAGCCTATGG
     3301 AAAAACGCCA GCAACGCGGC CTTTTTACGG TTCCTGGCCT TTTGCTGGCC TTTTGCTCAC
     3361 ATGTTCTTTC CTGCGTTATC CCCTGATTCT GTGGATAACC GTATTACCGC CTTTGAGTGA
     3421 GCTGATACCG CTCGCCGCAG CCGAACGACC GAGCGCAGCG AGTCAGTGAG CGAGGAAGCG
     3481 GAAGAGCGCC TGATGCGGTA TTTTCTCCTT ACGCATCTGT GCGGTATTTC ACACCGCATA
     3541 TATGGTGCAC TCTCAGTACA ATCTGCTCTG ATGCCGCATA GTTAAGCCAG TATACACTCC
     3601 GCTATCGCTA CGTGACTGGG TCATGGCTGC GCCCCGACAC CCGCCAACAC CCGCTGACGC
     3661 GCCCTGACGG GCTTGTCTGC TCCCGGCATC CGCTTACAGA CAAGCTGTGA CCGTCTCCGG
     3721 GAGCTGCATG TGTCAGAGGT TTTCACCGTC ATCACCGAAA CGCGCGAGGC AGCTGCGGTA
     3781 AAGCTCATCA GCGTGGTCGT GAAGCGATTC ACAGATGTCT GCCTGTTCAT CCGCGTCCAG
     3841 CTCGTTGAGT TTCTCCAGAA GCGTTAATGT CTGGCTTCTG ATAAAGCGGG CCATGTTAAG
     3901 GGCGGTTTTT TCCTGTTTGG TCACTGATGC CTCCGTGTAA GGGGGATTTC TGTTCATGGG
     3961 GGTAATGATA CCGATGAAAC GAGAGAGGAT GCTCACGATA CGGGTTACTG ATGATGAACA
     4021 TGCCCGGTTA CTGGAACGTT GTGAGGGTAA ACAACTGGCG GTATGGATGC GGCGGGACCA
     4081 GAGAAAAATC ACTCAGGGTC AATGCCAGCG CTTCGTTAAT ACAGATGTAG GTGTTCCACA
     4141 GGGTAGCCAG CAGCATCCTG CGATGCAGAT CCGGAACATA ATGGTGCAGG GCGCTGACTT
     4201 CCGCGTTTCC AGACTTTACG AAACACGGAA ACCGAAGACC ATTCATGTTG TTGCTCAGGT
     4261 CGCAGACGTT TTGCAGCAGC AGTCGCTTCA CGTTCGCTCG CGTATCGGTG ATTCATTCTG
     4321 CTAACCAGTA AGGCAACCCC GCCAGCCTAG CCGGGTCCTC AACGACAGGA GCACGATCAT
     4381 GCTAGTCATG CCCCGCGCCC ACCGGAAGGA GCTGACTGGG TTGAAGGCTC TCAAGGGCAT
     4441 CGGTCGAGAT CCCGGTGCCT AATGAGTGAG CTAACTTACA TTAATTGCGT TGCGCTCACT
     4501 GCCCGCTTTC CAGTCGGGAA ACCTGTCGTG CCAGCTGCAT TAATGAATCG GCCAACGCGC
     4561 GGGGAGAGGC GGTTTGCGTA TTGGGCGCCA GGGTGGTTTT TCTTTTCACC AGTGAGACGG
     4621 GCAACAGCTG ATTGCCCTTC ACCGCCTGGC CCTGAGAGAG TTGCAGCAAG CGGTCCACGC
     4681 TGGTTTGCCC CAGCAGGCGA AAATCCTGTT TGATGGTGGT TAACGGCGGG ATATAACATG
     4741 AGCTGTCTTC GGTATCGTCG TATCCCACTA CCGAGATGTC CGCACCAACG CGCAGCCCGG
     4801 ACTCGGTAAT GGCGCGCATT GCGCCCAGCG CCATCTGATC GTTGGCAACC AGCATCGCAG
     4861 TGGGAACGAT GCCCTCATTC AGCATTTGCA TGGTTTGTTG AAAACCGGAC ATGGCACTCC
     4921 AGTCGCCTTC CCGTTCCGCT ATCGGCTGAA TTTGATTGCG AGTGAGATAT TTATGCCAGC
     4981 CAGCCAGACG CAGACGCGCC GAGACAGAAC TTAATGGGCC CGCTAACAGC GCGATTTGCT
     5041 GGTGACCCAA TGCGACCAGA TGCTCCACGC CCAGTCGCGT ACCGTCTTCA TGGGAGAAAA
     5101 TAATACTGTT GATGGGTGTC TGGTCAGAGA CATCAAGAAA TAACGCCGGA ACATTAGTGC
     5161 AGGCAGCTTC CACAGCAATG GCATCCTGGT CATCCAGCGG ATAGTTAATG ATCAGCCCAC
     5221 TGACGCGTTG CGCGAGAAGA TTGTGCACCG CCGCTTTACA GGCTTCGACG CCGCTTCGTT
     5281 CTACCATCGA CACCACCACG CTGGCACCCA GTTGATCGGC GCGAGATTTA ATCGCCGCGA
     5341 CAATTTGCGA CGGCGCGTGC AGGGCCAGAC TGGAGGTGGC AACGCCAATC AGCAACGACT
     5401 GTTTGCCCGC CAGTTGTTGT GCCACGCGGT TGGGAATGTA ATTCAGCTCC GCCATCGCCG
     5461 CTTCCACTTT TTCCCGCGTT TTCGCAGAAA CGTGGCTGGC CTGGTTCACC ACGCGGGAAA
     5521 CGGTCTGATA AGAGACACCG GCATACTCTG CGACATCGTA TAACGTTACT GGTTTCACAT
     5581 TCACCACCCT GAATTGACTC TCTTCCGGGC GCTATCATGC CATACCGCGA AAGGTTTTGC
     5641 GCCATTCGAT GGTGTCCGGG ATCTCGACGC TCTCCCTTAT GCGACTCCTG CATTAGGAAG
     5701 CAGCCCAGTA GTAGGTTGAG GCCGTTGAGC ACCGCCGCCG CAAGGAATGG TGCATGCAAG
     5761 GAGATGGCGC CCAACAGTCC CCCGGCCACG GGGCCTGCCA CCATACCCAC GCCGAAACAA
     5821 GCGCTCATGA GCCCGAAGTG GCGAGCCCGA TCTTCCCCAT CGGTGATGTC GGCGATATAG
     5881 GCGCCAGCAA CCGCACCTGT GGCGCCGGTG ATGCCGGCCA CGATGCGTCC GGCGTAGAGG
     5941 ATCGAGATCG ATCTCGATCC CGCGAAAT
//