LOCUS       Copy_of_pET-6xHis-sfGFP 5896 bp ds-DNA     circular     24-FEB-2023
DEFINITION  .
FEATURES             Location/Qualifiers
     gene            complement(895..1146)
                     /label="tet (887 - 636)"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          1078..1097
                     /label="pBRrevBam_primer"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     promoter        1185..1201
                     /label="T7_promoter"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     promoter        1202..1203
                     /label="T7_promoter"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     regulatory      1203..1230
                     /label="lacO"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     feature         1248..1264
                     /label="T7_transl_en_RBS"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     gene            1272..2018
                     /label="sfGFP"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             1272..2018
                     /label="Translation 1272-2018"
                     /translation="MGHHHHHHGGASKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYK*"
     feature         1278..1295
                     /label="6xHIS"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     mutation        1317..1319
                     /label="E16R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1326..1328
                     /label="T19R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1332..1334
                     /label="V21K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1356..1358
                     /label="D29K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1395..1397
                     /label="E42K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1401..1403
                     /label="E44K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1416..1418
                     /label="N49R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     feature         1482..1511
                     /label="Y66 (EGFP)"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     mutation        1527..1529
                     /label="D86K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1539..1541
                     /label="Q90R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1569..1571
                     /label="E100K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1605..1607
                     /label="D112K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1614..1616
                     /label="T115K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1650..1652
                     /label="D127R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1671..1673
                     /label="E134K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1683..1685
                     /label="I138R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1698..1700
                     /label="D143K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1725..1727
                     /label="E152R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1746..1748
                     /label="N159K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1770..1772
                     /label="Q167R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1791..1793
                     /label="N174K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1815..1817
                     /label="E182K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1869..1871
                     /label="D200R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1890..1892
                     /label="D207R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1911..1913
                     /label="Q214R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1917..1919
                     /label="A216K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1935..1937
                     /label="N222K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1989..1991
                     /label="T240K"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        1998..2000
                     /label="M243R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     mutation        2007..2009
                     /label="L246R"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          complement(2125..2143)
                     /label="T7_Terminal_primer"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          complement(2793..2816)
                     /label="VY114"
                     /note="sequence: GCCTCACTGATTAAGCATTGGTAA"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     gene            complement(2793..3653)
                     /label="Ampicillin"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     promoter        complement(3696..3724)
                     /label="AmpR_promoter"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     origin          3811..4430
                     /label="pBR322_origin"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          4829..4851
                     /label="pGEX_3_primer"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     feature         complement(4845..5036)
                     /label="ROP"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     regulatory      complement(5608..803)
                     /label="lacI"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
ORIGIN
        1 CCAACGCGCA GCCCGGACTC GGTAATGGCG CGCATTGCGC CCAGCGCCAT CTGATCGTTG
       61 GCAACCAGCA TCGCAGTGGG AACGATGCCC TCATTCAGCA TTTGCATGGT TTGTTGAAAA
      121 CCGGACATGG CACTCCAGTC GCCTTCCCGT TCCGCTATCG GCTGAATTTG ATTGCGAGTG
      181 AGATATTTAT GCCAGCCAGC CAGACGCAGA CGCGCCGAGA CAGAACTTAA TGGGCCCGCT
      241 AACAGCGCGA TTTGCTGGTG ACCCAATGCG ACCAGATGCT CCACGCCCAG TCGCGTACCG
      301 TCTTCATGGG AGAAAATAAT ACTGTTGATG GGTGTCTGGT CAGAGACATC AAGAAATAAC
      361 GCCGGAACAT TAGTGCAGGC AGCTTCCACA GCAATGGCAT CCTGGTCATC CAGCGGATAG
      421 TTAATGATCA GCCCACTGAC GCGTTGCGCG AGAAGATTGT GCACCGCCGC TTTACAGGCT
      481 TCGACGCCGC TTCGTTCTAC CATCGACACC ACCACGCTGG CACCCAGTTG ATCGGCGCGA
      541 GATTTAATCG CCGCGACAAT TTGCGACGGC GCGTGCAGGG CCAGACTGGA GGTGGCAACG
      601 CCAATCAGCA ACGACTGTTT GCCCGCCAGT TGTTGTGCCA CGCGGTTGGG AATGTAATTC
      661 AGCTCCGCCA TCGCCGCTTC CACTTTTTCC CGCGTTTTCG CAGAAACGTG GCTGGCCTGG
      721 TTCACCACGC GGGAAACGGT CTGATAAGAG ACACCGGCAT ACTCTGCGAC ATCGTATAAC
      781 GTTACTGGTT TCACATTCAC CACCCTGAAT TGACTCTCTT CCGGGCGCTA TCATGCCATA
      841 CCGCGAAAGG TTTTGCGCCA TTCGATGGTG TCCGGGATCT CGACGCTCTC CCTTATGCGA
      901 CTCCTGCATT AGGAAGCAGC CCAGTAGTAG GTTGAGGCCG TTGAGCACCG CCGCCGCAAG
      961 GAATGGTGCA TGCAAGGAGA TGGCGCCCAA CAGTCCCCCG GCCACGGGGC CTGCCACCAT
     1021 ACCCACGCCG AAACAAGCGC TCATGAGCCC GAAGTGGCGA GCCCGATCTT CCCCATCGGT
     1081 GATGTCGGCG ATATAGGCGC CAGCAACCGC ACCTGTGGCG CCGGTGATGC CGGCCACGAT
     1141 GCGTCCGGCG TAGAGGATCG AGATCGATCT CGATCCCGCG AAATTAATAC GACTCACTAT
     1201 AGGGGAATTG TGAGCGGATA ACAATTCCCC TCTAGAAATA ATTTTGTTTA ACTTTAAGAA
     1261 GGAGATATAC CATGGGTCAT CACCACCACC ATCACGGTGG CGCTAGCAAA GGTGAAGAGC
     1321 TGTTTACGGG TGTAGTACCG ATCTTAGTGG AATTAGACGG CGACGTGAAC GGTCACAAAT
     1381 TTAGCGTGCG CGGCGAAGGC GAAGGTGACG CTACCAATGG TAAATTGACC CTGAAGTTTA
     1441 TTTGCACAAC AGGCAAATTA CCCGTTCCGT GGCCCACCTT AGTGACCACC CTGACCTATG
     1501 GCGTTCAGTG CTTCAGTCGT TACCCAGATC ATATGAAACA ACACGATTTT TTCAAATCAG
     1561 CCATGCCTGA AGGATATGTT CAAGAGCGTA CAATCAGCTT CAAGGACGAT GGCACCTATA
     1621 AAACGCGTGC GGAAGTGAAA TTTGAAGGCG ACACATTAGT AAACCGTATC GAACTGAAAG
     1681 GTATCGACTT CAAAGAAGAC GGCAACATTT TAGGCCATAA GCTGGAATAT AACTTTAATT
     1741 CTCATAACGT GTATATTACG GCCGATAAAC AGAAAAACGG TATCAAGGCA AATTTCAAAA
     1801 TTCGCCATAA CGTGGAAGAC GGCAGCGTTC AATTAGCGGA TCATTATCAA CAAAACACGC
     1861 CGATTGGTGA CGGGCCTGTA CTGTTACCTG ACAACCACTA CCTGAGCACC CAGTCAGCAC
     1921 TGAGCAAAGA TCCGAACGAA AAACGCGATC ACATGGTTCT GTTAGAATTC GTGACCGCTG
     1981 CAGGCATTAC TCACGGAATG GACGAACTCT ACAAGTAATA ATGAGGTACC CTCGAGTCTG
     2041 GTAAAGAAAC CGCTGCTGCG AAATTTGAAC GCCAGCACAT GGACTCGTCT ACTAGCGCAG
     2101 CTTAATTAAC CTAGGCTGCT GCCACCGCTG AGCAATAACT AGCATAACCC CTTGGGGCCT
     2161 CTAAACGGGT CTTGAGGGGT TTTTTGCTGA AAGGAGGAAC TATATCCGGA TTGGCGAATG
     2221 GGACGCGCCC TGTAGCGGCG CATTAAGCGC GGCGGGTGTG GTGGTTACGC GCAGCGTGAC
     2281 CGCTACACTT GCCAGCGCCC TAGCGCCCGC TCCTTTCGCT TTCTTCCCTT CCTTTCTCGC
     2341 CACGTTCGCC GGCTTTCCCC GTCAAGCTCT AAATCGGGGG CTCCCTTTAG GGTTCCGATT
     2401 TAGTGCTTTA CGGCACCTCG ACCCCAAAAA ACTTGATTAG GGTGATGGTT CACGTAGTGG
     2461 GCCATCGCCC TGATAGACGG TTTTTCGCCC TTTGACGTTG GAGTCCACGT TCTTTAATAG
     2521 TGGACTCTTG TTCCAAACTG GAACAACACT CAACCCTATC TCGGTCTATT CTTTTGATTT
     2581 ATAAGGGATT TTGCCGATTT CGGCCTATTG GTTAAAAAAT GAGCTGATTT AACAAAAATT
     2641 TAACGCGAAT TTTAACAAAA TATTAACGTT TACAATTTCT GGCGGCACGA TGGCATGAGA
     2701 TTATCAAAAA GGATCTTCAC CTAGATCCTT TTAAATTAAA AATGAAGTTT TAAATCAATC
     2761 TAAAGTATAT ATGAGTAAAC TTGGTCTGAC AGTTACCAAT GCTTAATCAG TGAGGCACCT
     2821 ATCTCAGCGA TCTGTCTATT TCGTTCATCC ATAGTTGCCT GACTCCCCGT CGTGTAGATA
     2881 ACTACGATAC GGGAGGGCTT ACCATCTGGC CCCAGTGCTG CAATGATACC GCGAGACCCA
     2941 CGCTCACCGG CTCCAGATTT ATCAGCAATA AACCAGCCAG CCGGAAGGGC CGAGCGCAGA
     3001 AGTGGTCCTG CAACTTTATC CGCCTCCATC CAGTCTATTA ATTGTTGCCG GGAAGCTAGA
     3061 GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC AACGTTGTTG CCATTGCTAC AGGCATCGTG
     3121 GTGTCACGCT CGTCGTTTGG TATGGCTTCA TTCAGCTCCG GTTCCCAACG ATCAAGGCGA
     3181 GTTACATGAT CCCCCATGTT GTGCAAAAAA GCGGTTAGCT CCTTCGGTCC TCCGATCGTT
     3241 GTCAGAAGTA AGTTGGCCGC AGTGTTATCA CTCATGGTTA TGGCAGCACT GCATAATTCT
     3301 CTTACTGTCA TGCCATCCGT AAGATGCTTT TCTGTGACTG GTGAGTACTC AACCAAGTCA
     3361 TTCTGAGAAT AGTGTATGCG GCGACCGAGT TGCTCTTGCC CGGCGTCAAT ACGGGATAAT
     3421 ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG CTCATCATTG GAAAACGTTC TTCGGGGCGA
     3481 AAACTCTCAA GGATCTTACC GCTGTTGAGA TCCAGTTCGA TGTAACCCAC TCGTGCACCC
     3541 AACTGATCTT CAGCATCTTT TACTTTCACC AGCGTTTCTG GGTGAGCAAA AACAGGAAGG
     3601 CAAAATGCCG CAAAAAAGGG AATAAGGGCG ACACGGAAAT GTTGAATACT CATACTCTTC
     3661 CTTTTTCAAT CATGATTGAA GCATTTATCA GGGTTATTGT CTCATGAGCG GATACATATT
     3721 TGAATGTATT TAGAAAAATA AACAAATAGG TCATGACCAA AATCCCTTAA CGTGAGTTTT
     3781 CGTTCCACTG AGCGTCAGAC CCCGTAGAAA AGATCAAAGG ATCTTCTTGA GATCCTTTTT
     3841 TTCTGCGCGT AATCTGCTGC TTGCAAACAA AAAAACCACC GCTACCAGCG GTGGTTTGTT
     3901 TGCCGGATCA AGAGCTACCA ACTCTTTTTC CGAAGGTAAC TGGCTTCAGC AGAGCGCAGA
     3961 TACCAAATAC TGTCCTTCTA GTGTAGCCGT AGTTAGGCCA CCACTTCAAG AACTCTGTAG
     4021 CACCGCCTAC ATACCTCGCT CTGCTAATCC TGTTACCAGT GGCTGCTGCC AGTGGCGATA
     4081 AGTCGTGTCT TACCGGGTTG GACTCAAGAC GATAGTTACC GGATAAGGCG CAGCGGTCGG
     4141 GCTGAACGGG GGGTTCGTGC ACACAGCCCA GCTTGGAGCG AACGACCTAC ACCGAACTGA
     4201 GATACCTACA GCGTGAGCTA TGAGAAAGCG CCACGCTTCC CGAAGGGAGA AAGGCGGACA
     4261 GGTATCCGGT AAGCGGCAGG GTCGGAACAG GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA
     4321 ACGCCTGGTA TCTTTATAGT CCTGTCGGGT TTCGCCACCT CTGACTTGAG CGTCGATTTT
     4381 TGTGATGCTC GTCAGGGGGG CGGAGCCTAT GGAAAAACGC CAGCAACGCG GCCTTTTTAC
     4441 GGTTCCTGGC CTTTTGCTGG CCTTTTGCTC ACATGTTCTT TCCTGCGTTA TCCCCTGATT
     4501 CTGTGGATAA CCGTATTACC GCCTTTGAGT GAGCTGATAC CGCTCGCCGC AGCCGAACGA
     4561 CCGAGCGCAG CGAGTCAGTG AGCGAGGAAG CGGAAGAGCG CCTGATGCGG TATTTTCTCC
     4621 TTACGCATCT GTGCGGTATT TCACACCGCA TATATGGTGC ACTCTCAGTA CAATCTGCTC
     4681 TGATGCCGCA TAGTTAAGCC AGTATACACT CCGCTATCGC TACGTGACTG GGTCATGGCT
     4741 GCGCCCCGAC ACCCGCCAAC ACCCGCTGAC GCGCCCTGAC GGGCTTGTCT GCTCCCGGCA
     4801 TCCGCTTACA GACAAGCTGT GACCGTCTCC GGGAGCTGCA TGTGTCAGAG GTTTTCACCG
     4861 TCATCACCGA AACGCGCGAG GCAGCTGCGG TAAAGCTCAT CAGCGTGGTC GTGAAGCGAT
     4921 TCACAGATGT CTGCCTGTTC ATCCGCGTCC AGCTCGTTGA GTTTCTCCAG AAGCGTTAAT
     4981 GTCTGGCTTC TGATAAAGCG GGCCATGTTA AGGGCGGTTT TTTCCTGTTT GGTCACTGAT
     5041 GCCTCCGTGT AAGGGGGATT TCTGTTCATG GGGGTAATGA TACCGATGAA ACGAGAGAGG
     5101 ATGCTCACGA TACGGGTTAC TGATGATGAA CATGCCCGGT TACTGGAACG TTGTGAGGGT
     5161 AAACAACTGG CGGTATGGAT GCGGCGGGAC CAGAGAAAAA TCACTCAGGG TCAATGCCAG
     5221 CGCTTCGTTA ATACAGATGT AGGTGTTCCA CAGGGTAGCC AGCAGCATCC TGCGATGCAG
     5281 ATCCGGAACA TAATGGTGCA GGGCGCTGAC TTCCGCGTTT CCAGACTTTA CGAAACACGG
     5341 AAACCGAAGA CCATTCATGT TGTTGCTCAG GTCGCAGACG TTTTGCAGCA GCAGTCGCTT
     5401 CACGTTCGCT CGCGTATCGG TGATTCATTC TGCTAACCAG TAAGGCAACC CCGCCAGCCT
     5461 AGCCGGGTCC TCAACGACAG GAGCACGATC ATGCTAGTCA TGCCCCGCGC CCACCGGAAG
     5521 GAGCTGACTG GGTTGAAGGC TCTCAAGGGC ATCGGTCGAG ATCCCGGTGC CTAATGAGTG
     5581 AGCTAACTTA CATTAATTGC GTTGCGCTCA CTGCCCGCTT TCCAGTCGGG AAACCTGTCG
     5641 TGCCAGCTGC ATTAATGAAT CGGCCAACGC GCGGGGAGAG GCGGTTTGCG TATTGGGCGC
     5701 CAGGGTGGTT TTTCTTTTCA CCAGTGAGAC GGGCAACAGC TGATTGCCCT TCACCGCCTG
     5761 GCCCTGAGAG AGTTGCAGCA AGCGGTCCAC GCTGGTTTGC CCCAGCAGGC GAAAATCCTG
     5821 TTTGATGGTG GTTAACGGCG GGATATAACA TGAGCTGTCT TCGGTATCGT CGTATCCCAC
     5881 TACCGAGATG TCCGCA
//