LOCUS       04_pET-6xHis-GFP(-6)-H1 5908 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..858
                     /label="6xHis-GFP(-6)-H1"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /translation="MGHHHHHHGGASKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKKKRRRKK**"
     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..852
                     /label="H1 Peptide"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(953..971)
                     /label="T7_Terminal_primer"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     gene            complement(1621..2481)
                     /label="Ampicillin"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     promoter        complement(2524..2552)
                     /label="AmpR_promoter"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     origin          2639..3258
                     /label="pBR322_origin"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          3657..3679
                     /label="pGEX_3_primer"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     feature         complement(3673..3864)
                     /label="ROP"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     regulatory      complement(4436..5527)
                     /label="lacI"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     gene            complement(5619..5870)
                     /label="tet (887 - 636)"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          5802..5821
                     /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 GAAAAAGAGG
      841 AGAAGGAAGA AATAATAACT CGAGTCTGGT AAAGAAACCG CTGCTGCGAA ATTTGAACGC
      901 CAGCACATGG ACTCGTCTAC TAGCGCAGCT TAATTAACCT AGGCTGCTGC CACCGCTGAG
      961 CAATAACTAG CATAACCCCT TGGGGCCTCT AAACGGGTCT TGAGGGGTTT TTTGCTGAAA
     1021 GGAGGAACTA TATCCGGATT GGCGAATGGG ACGCGCCCTG TAGCGGCGCA TTAAGCGCGG
     1081 CGGGTGTGGT GGTTACGCGC AGCGTGACCG CTACACTTGC CAGCGCCCTA GCGCCCGCTC
     1141 CTTTCGCTTT CTTCCCTTCC TTTCTCGCCA CGTTCGCCGG CTTTCCCCGT CAAGCTCTAA
     1201 ATCGGGGGCT CCCTTTAGGG TTCCGATTTA GTGCTTTACG GCACCTCGAC CCCAAAAAAC
     1261 TTGATTAGGG TGATGGTTCA CGTAGTGGGC CATCGCCCTG ATAGACGGTT TTTCGCCCTT
     1321 TGACGTTGGA GTCCACGTTC TTTAATAGTG GACTCTTGTT CCAAACTGGA ACAACACTCA
     1381 ACCCTATCTC GGTCTATTCT TTTGATTTAT AAGGGATTTT GCCGATTTCG GCCTATTGGT
     1441 TAAAAAATGA GCTGATTTAA CAAAAATTTA ACGCGAATTT TAACAAAATA TTAACGTTTA
     1501 CAATTTCTGG CGGCACGATG GCATGAGATT ATCAAAAAGG ATCTTCACCT AGATCCTTTT
     1561 AAATTAAAAA TGAAGTTTTA AATCAATCTA AAGTATATAT GAGTAAACTT GGTCTGACAG
     1621 TTACCAATGC TTAATCAGTG AGGCACCTAT CTCAGCGATC TGTCTATTTC GTTCATCCAT
     1681 AGTTGCCTGA CTCCCCGTCG TGTAGATAAC TACGATACGG GAGGGCTTAC CATCTGGCCC
     1741 CAGTGCTGCA ATGATACCGC GAGACCCACG CTCACCGGCT CCAGATTTAT CAGCAATAAA
     1801 CCAGCCAGCC GGAAGGGCCG AGCGCAGAAG TGGTCCTGCA ACTTTATCCG CCTCCATCCA
     1861 GTCTATTAAT TGTTGCCGGG AAGCTAGAGT AAGTAGTTCG CCAGTTAATA GTTTGCGCAA
     1921 CGTTGTTGCC ATTGCTACAG GCATCGTGGT GTCACGCTCG TCGTTTGGTA TGGCTTCATT
     1981 CAGCTCCGGT TCCCAACGAT CAAGGCGAGT TACATGATCC CCCATGTTGT GCAAAAAAGC
     2041 GGTTAGCTCC TTCGGTCCTC CGATCGTTGT CAGAAGTAAG TTGGCCGCAG TGTTATCACT
     2101 CATGGTTATG GCAGCACTGC ATAATTCTCT TACTGTCATG CCATCCGTAA GATGCTTTTC
     2161 TGTGACTGGT GAGTACTCAA CCAAGTCATT CTGAGAATAG TGTATGCGGC GACCGAGTTG
     2221 CTCTTGCCCG GCGTCAATAC GGGATAATAC CGCGCCACAT AGCAGAACTT TAAAAGTGCT
     2281 CATCATTGGA AAACGTTCTT CGGGGCGAAA ACTCTCAAGG ATCTTACCGC TGTTGAGATC
     2341 CAGTTCGATG TAACCCACTC GTGCACCCAA CTGATCTTCA GCATCTTTTA CTTTCACCAG
     2401 CGTTTCTGGG TGAGCAAAAA CAGGAAGGCA AAATGCCGCA AAAAAGGGAA TAAGGGCGAC
     2461 ACGGAAATGT TGAATACTCA TACTCTTCCT TTTTCAATCA TGATTGAAGC ATTTATCAGG
     2521 GTTATTGTCT CATGAGCGGA TACATATTTG AATGTATTTA GAAAAATAAA CAAATAGGTC
     2581 ATGACCAAAA TCCCTTAACG TGAGTTTTCG TTCCACTGAG CGTCAGACCC CGTAGAAAAG
     2641 ATCAAAGGAT CTTCTTGAGA TCCTTTTTTT CTGCGCGTAA TCTGCTGCTT GCAAACAAAA
     2701 AAACCACCGC TACCAGCGGT GGTTTGTTTG CCGGATCAAG AGCTACCAAC TCTTTTTCCG
     2761 AAGGTAACTG GCTTCAGCAG AGCGCAGATA CCAAATACTG TCCTTCTAGT GTAGCCGTAG
     2821 TTAGGCCACC ACTTCAAGAA CTCTGTAGCA CCGCCTACAT ACCTCGCTCT GCTAATCCTG
     2881 TTACCAGTGG CTGCTGCCAG TGGCGATAAG TCGTGTCTTA CCGGGTTGGA CTCAAGACGA
     2941 TAGTTACCGG ATAAGGCGCA GCGGTCGGGC TGAACGGGGG GTTCGTGCAC ACAGCCCAGC
     3001 TTGGAGCGAA CGACCTACAC CGAACTGAGA TACCTACAGC GTGAGCTATG AGAAAGCGCC
     3061 ACGCTTCCCG AAGGGAGAAA GGCGGACAGG TATCCGGTAA GCGGCAGGGT CGGAACAGGA
     3121 GAGCGCACGA GGGAGCTTCC AGGGGGAAAC GCCTGGTATC TTTATAGTCC TGTCGGGTTT
     3181 CGCCACCTCT GACTTGAGCG TCGATTTTTG TGATGCTCGT CAGGGGGGCG GAGCCTATGG
     3241 AAAAACGCCA GCAACGCGGC CTTTTTACGG TTCCTGGCCT TTTGCTGGCC TTTTGCTCAC
     3301 ATGTTCTTTC CTGCGTTATC CCCTGATTCT GTGGATAACC GTATTACCGC CTTTGAGTGA
     3361 GCTGATACCG CTCGCCGCAG CCGAACGACC GAGCGCAGCG AGTCAGTGAG CGAGGAAGCG
     3421 GAAGAGCGCC TGATGCGGTA TTTTCTCCTT ACGCATCTGT GCGGTATTTC ACACCGCATA
     3481 TATGGTGCAC TCTCAGTACA ATCTGCTCTG ATGCCGCATA GTTAAGCCAG TATACACTCC
     3541 GCTATCGCTA CGTGACTGGG TCATGGCTGC GCCCCGACAC CCGCCAACAC CCGCTGACGC
     3601 GCCCTGACGG GCTTGTCTGC TCCCGGCATC CGCTTACAGA CAAGCTGTGA CCGTCTCCGG
     3661 GAGCTGCATG TGTCAGAGGT TTTCACCGTC ATCACCGAAA CGCGCGAGGC AGCTGCGGTA
     3721 AAGCTCATCA GCGTGGTCGT GAAGCGATTC ACAGATGTCT GCCTGTTCAT CCGCGTCCAG
     3781 CTCGTTGAGT TTCTCCAGAA GCGTTAATGT CTGGCTTCTG ATAAAGCGGG CCATGTTAAG
     3841 GGCGGTTTTT TCCTGTTTGG TCACTGATGC CTCCGTGTAA GGGGGATTTC TGTTCATGGG
     3901 GGTAATGATA CCGATGAAAC GAGAGAGGAT GCTCACGATA CGGGTTACTG ATGATGAACA
     3961 TGCCCGGTTA CTGGAACGTT GTGAGGGTAA ACAACTGGCG GTATGGATGC GGCGGGACCA
     4021 GAGAAAAATC ACTCAGGGTC AATGCCAGCG CTTCGTTAAT ACAGATGTAG GTGTTCCACA
     4081 GGGTAGCCAG CAGCATCCTG CGATGCAGAT CCGGAACATA ATGGTGCAGG GCGCTGACTT
     4141 CCGCGTTTCC AGACTTTACG AAACACGGAA ACCGAAGACC ATTCATGTTG TTGCTCAGGT
     4201 CGCAGACGTT TTGCAGCAGC AGTCGCTTCA CGTTCGCTCG CGTATCGGTG ATTCATTCTG
     4261 CTAACCAGTA AGGCAACCCC GCCAGCCTAG CCGGGTCCTC AACGACAGGA GCACGATCAT
     4321 GCTAGTCATG CCCCGCGCCC ACCGGAAGGA GCTGACTGGG TTGAAGGCTC TCAAGGGCAT
     4381 CGGTCGAGAT CCCGGTGCCT AATGAGTGAG CTAACTTACA TTAATTGCGT TGCGCTCACT
     4441 GCCCGCTTTC CAGTCGGGAA ACCTGTCGTG CCAGCTGCAT TAATGAATCG GCCAACGCGC
     4501 GGGGAGAGGC GGTTTGCGTA TTGGGCGCCA GGGTGGTTTT TCTTTTCACC AGTGAGACGG
     4561 GCAACAGCTG ATTGCCCTTC ACCGCCTGGC CCTGAGAGAG TTGCAGCAAG CGGTCCACGC
     4621 TGGTTTGCCC CAGCAGGCGA AAATCCTGTT TGATGGTGGT TAACGGCGGG ATATAACATG
     4681 AGCTGTCTTC GGTATCGTCG TATCCCACTA CCGAGATGTC CGCACCAACG CGCAGCCCGG
     4741 ACTCGGTAAT GGCGCGCATT GCGCCCAGCG CCATCTGATC GTTGGCAACC AGCATCGCAG
     4801 TGGGAACGAT GCCCTCATTC AGCATTTGCA TGGTTTGTTG AAAACCGGAC ATGGCACTCC
     4861 AGTCGCCTTC CCGTTCCGCT ATCGGCTGAA TTTGATTGCG AGTGAGATAT TTATGCCAGC
     4921 CAGCCAGACG CAGACGCGCC GAGACAGAAC TTAATGGGCC CGCTAACAGC GCGATTTGCT
     4981 GGTGACCCAA TGCGACCAGA TGCTCCACGC CCAGTCGCGT ACCGTCTTCA TGGGAGAAAA
     5041 TAATACTGTT GATGGGTGTC TGGTCAGAGA CATCAAGAAA TAACGCCGGA ACATTAGTGC
     5101 AGGCAGCTTC CACAGCAATG GCATCCTGGT CATCCAGCGG ATAGTTAATG ATCAGCCCAC
     5161 TGACGCGTTG CGCGAGAAGA TTGTGCACCG CCGCTTTACA GGCTTCGACG CCGCTTCGTT
     5221 CTACCATCGA CACCACCACG CTGGCACCCA GTTGATCGGC GCGAGATTTA ATCGCCGCGA
     5281 CAATTTGCGA CGGCGCGTGC AGGGCCAGAC TGGAGGTGGC AACGCCAATC AGCAACGACT
     5341 GTTTGCCCGC CAGTTGTTGT GCCACGCGGT TGGGAATGTA ATTCAGCTCC GCCATCGCCG
     5401 CTTCCACTTT TTCCCGCGTT TTCGCAGAAA CGTGGCTGGC CTGGTTCACC ACGCGGGAAA
     5461 CGGTCTGATA AGAGACACCG GCATACTCTG CGACATCGTA TAACGTTACT GGTTTCACAT
     5521 TCACCACCCT GAATTGACTC TCTTCCGGGC GCTATCATGC CATACCGCGA AAGGTTTTGC
     5581 GCCATTCGAT GGTGTCCGGG ATCTCGACGC TCTCCCTTAT GCGACTCCTG CATTAGGAAG
     5641 CAGCCCAGTA GTAGGTTGAG GCCGTTGAGC ACCGCCGCCG CAAGGAATGG TGCATGCAAG
     5701 GAGATGGCGC CCAACAGTCC CCCGGCCACG GGGCCTGCCA CCATACCCAC GCCGAAACAA
     5761 GCGCTCATGA GCCCGAAGTG GCGAGCCCGA TCTTCCCCAT CGGTGATGTC GGCGATATAG
     5821 GCGCCAGCAA CCGCACCTGT GGCGCCGGTG ATGCCGGCCA CGATGCGTCC GGCGTAGAGG
     5881 ATCGAGATCG ATCTCGATCC CGCGAAAT
//