LOCUS       pVY-3:_sfGFP-tag3       5971 bp ds-DNA     circular     05-OCT-2022
DEFINITION  .
FEATURES             Location/Qualifiers
     primer          158..180
                     /label="pGEX_3_primer"
     feature         complement(174..365)
                     /label="ROP"
     regulatory      complement(937..2028)
                     /label="lacI"
     gene            complement(2120..2371)
                     /label="tet (887 - 636)"
     primer          2303..2322
                     /label="pBRrevBam_primer"
     promoter        2410..2426
                     /label="T7_promoter"
     promoter        2427..2428
                     /label="T7_promoter"
     regulatory      2428..2455
                     /label="lacO"
     feature         2473..2489
                     /label="T7_transl_en_RBS"
     gene            2497..3327
                     /label="sfGFP-tag3"
     CDS             2497..3330
                     /label="Translation 2497-3330"
                     /translation="MGHHHHHHGGASKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDGTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKKGGSKKRKKRGGSKKRKKRGGSKKRKKR**"
     feature         2503..2520
                     /label="6xHIS"
     mutation        2542..2544
                     /label="E16R"
     mutation        2551..2553
                     /label="T19R"
     mutation        2557..2559
                     /label="V21K"
     mutation        2581..2583
                     /label="D29K"
     mutation        2620..2622
                     /label="E42K"
     mutation        2626..2628
                     /label="E44K"
     mutation        2641..2643
                     /label="N49R"
     feature         2707..2736
                     /label="Y66 (EGFP)"
     mutation        2752..2754
                     /label="D86K"
     mutation        2764..2766
                     /label="Q90R"
     mutation        2794..2796
                     /label="E100K"
     mutation        2830..2832
                     /label="D112K"
     mutation        2839..2841
                     /label="T115K"
     mutation        2875..2877
                     /label="D127R"
     mutation        2896..2898
                     /label="E134K"
     mutation        2908..2910
                     /label="I138R"
     mutation        2923..2925
                     /label="D143K"
     mutation        2950..2952
                     /label="E152R"
     mutation        2971..2973
                     /label="N159K"
     mutation        2995..2997
                     /label="Q167R"
     mutation        3016..3018
                     /label="N174K"
     mutation        3040..3042
                     /label="E182K"
     mutation        3094..3096
                     /label="D200R"
     mutation        3115..3117
                     /label="D207R"
     mutation        3136..3138
                     /label="Q214R"
     mutation        3142..3144
                     /label="A216K"
     mutation        3160..3162
                     /label="N222K"
     mutation        3214..3216
                     /label="T240K"
     mutation        3223..3225
                     /label="M243R"
     mutation        3232..3234
                     /label="L246R"
     misc_feature    3241..3324
                     /label="tag3"
     primer          complement(3249..3339)
                     /label="sfGFP-posB4-XhoI-reverse"
                     /note="sequence: ATACTCGAGTTATTAACGCTTCTTACGCTTTTTGCTACCGCCACGCTTCTTACGCTTTTTGCTACCGCCTCTTTTCTTCCTTTTCTTTGAA"
     primer          complement(3425..3443)
                     /label="T7_Terminal_primer"
     gene            complement(4093..4953)
                     /label="Ampicillin"
     promoter        complement(4996..5024)
                     /label="AmpR_promoter"
     origin          5111..5730
                     /label="pBR322_origin"
ORIGIN
        1 AATCTGCTCT GATGCCGCAT AGTTAAGCCA GTATACACTC CGCTATCGCT ACGTGACTGG
       61 GTCATGGCTG CGCCCCGACA CCCGCCAACA CCCGCTGACG CGCCCTGACG GGCTTGTCTG
      121 CTCCCGGCAT CCGCTTACAG ACAAGCTGTG ACCGTCTCCG GGAGCTGCAT GTGTCAGAGG
      181 TTTTCACCGT CATCACCGAA ACGCGCGAGG CAGCTGCGGT AAAGCTCATC AGCGTGGTCG
      241 TGAAGCGATT CACAGATGTC TGCCTGTTCA TCCGCGTCCA GCTCGTTGAG TTTCTCCAGA
      301 AGCGTTAATG TCTGGCTTCT GATAAAGCGG GCCATGTTAA GGGCGGTTTT TTCCTGTTTG
      361 GTCACTGATG CCTCCGTGTA AGGGGGATTT CTGTTCATGG GGGTAATGAT ACCGATGAAA
      421 CGAGAGAGGA TGCTCACGAT ACGGGTTACT GATGATGAAC ATGCCCGGTT ACTGGAACGT
      481 TGTGAGGGTA AACAACTGGC GGTATGGATG CGGCGGGACC AGAGAAAAAT CACTCAGGGT
      541 CAATGCCAGC GCTTCGTTAA TACAGATGTA GGTGTTCCAC AGGGTAGCCA GCAGCATCCT
      601 GCGATGCAGA TCCGGAACAT AATGGTGCAG GGCGCTGACT TCCGCGTTTC CAGACTTTAC
      661 GAAACACGGA AACCGAAGAC CATTCATGTT GTTGCTCAGG TCGCAGACGT TTTGCAGCAG
      721 CAGTCGCTTC ACGTTCGCTC GCGTATCGGT GATTCATTCT GCTAACCAGT AAGGCAACCC
      781 CGCCAGCCTA GCCGGGTCCT CAACGACAGG AGCACGATCA TGCTAGTCAT GCCCCGCGCC
      841 CACCGGAAGG AGCTGACTGG GTTGAAGGCT CTCAAGGGCA TCGGTCGAGA TCCCGGTGCC
      901 TAATGAGTGA GCTAACTTAC ATTAATTGCG TTGCGCTCAC TGCCCGCTTT CCAGTCGGGA
      961 AACCTGTCGT GCCAGCTGCA TTAATGAATC GGCCAACGCG CGGGGAGAGG CGGTTTGCGT
     1021 ATTGGGCGCC AGGGTGGTTT TTCTTTTCAC CAGTGAGACG GGCAACAGCT GATTGCCCTT
     1081 CACCGCCTGG CCCTGAGAGA GTTGCAGCAA GCGGTCCACG CTGGTTTGCC CCAGCAGGCG
     1141 AAAATCCTGT TTGATGGTGG TTAACGGCGG GATATAACAT GAGCTGTCTT CGGTATCGTC
     1201 GTATCCCACT ACCGAGATGT CCGCACCAAC GCGCAGCCCG GACTCGGTAA TGGCGCGCAT
     1261 TGCGCCCAGC GCCATCTGAT CGTTGGCAAC CAGCATCGCA GTGGGAACGA TGCCCTCATT
     1321 CAGCATTTGC ATGGTTTGTT GAAAACCGGA CATGGCACTC CAGTCGCCTT CCCGTTCCGC
     1381 TATCGGCTGA ATTTGATTGC GAGTGAGATA TTTATGCCAG CCAGCCAGAC GCAGACGCGC
     1441 CGAGACAGAA CTTAATGGGC CCGCTAACAG CGCGATTTGC TGGTGACCCA ATGCGACCAG
     1501 ATGCTCCACG CCCAGTCGCG TACCGTCTTC ATGGGAGAAA ATAATACTGT TGATGGGTGT
     1561 CTGGTCAGAG ACATCAAGAA ATAACGCCGG AACATTAGTG CAGGCAGCTT CCACAGCAAT
     1621 GGCATCCTGG TCATCCAGCG GATAGTTAAT GATCAGCCCA CTGACGCGTT GCGCGAGAAG
     1681 ATTGTGCACC GCCGCTTTAC AGGCTTCGAC GCCGCTTCGT TCTACCATCG ACACCACCAC
     1741 GCTGGCACCC AGTTGATCGG CGCGAGATTT AATCGCCGCG ACAATTTGCG ACGGCGCGTG
     1801 CAGGGCCAGA CTGGAGGTGG CAACGCCAAT CAGCAACGAC TGTTTGCCCG CCAGTTGTTG
     1861 TGCCACGCGG TTGGGAATGT AATTCAGCTC CGCCATCGCC GCTTCCACTT TTTCCCGCGT
     1921 TTTCGCAGAA ACGTGGCTGG CCTGGTTCAC CACGCGGGAA ACGGTCTGAT AAGAGACACC
     1981 GGCATACTCT GCGACATCGT ATAACGTTAC TGGTTTCACA TTCACCACCC TGAATTGACT
     2041 CTCTTCCGGG CGCTATCATG CCATACCGCG AAAGGTTTTG CGCCATTCGA TGGTGTCCGG
     2101 GATCTCGACG CTCTCCCTTA TGCGACTCCT GCATTAGGAA GCAGCCCAGT AGTAGGTTGA
     2161 GGCCGTTGAG CACCGCCGCC GCAAGGAATG GTGCATGCAA GGAGATGGCG CCCAACAGTC
     2221 CCCCGGCCAC GGGGCCTGCC ACCATACCCA CGCCGAAACA AGCGCTCATG AGCCCGAAGT
     2281 GGCGAGCCCG ATCTTCCCCA TCGGTGATGT CGGCGATATA GGCGCCAGCA ACCGCACCTG
     2341 TGGCGCCGGT GATGCCGGCC ACGATGCGTC CGGCGTAGAG GATCGAGATC GATCTCGATC
     2401 CCGCGAAATT AATACGACTC ACTATAGGGG AATTGTGAGC GGATAACAAT TCCCCTCTAG
     2461 AAATAATTTT GTTTAACTTT AAGAAGGAGA TATACCATGG GTCATCACCA CCACCATCAC
     2521 GGTGGCGCTA GCAAAGGTGA AGAGCTGTTT ACGGGTGTAG TACCGATCTT AGTGGAATTA
     2581 GACGGCGACG TGAACGGTCA CAAATTTAGC GTGCGCGGCG AAGGCGAAGG TGACGCTACC
     2641 AATGGTAAAT TGACCCTGAA GTTTATTTGC ACAACAGGCA AATTACCCGT TCCGTGGCCC
     2701 ACCTTAGTGA CCACCCTGAC CTATGGCGTT CAGTGCTTCA GTCGTTACCC AGATCATATG
     2761 AAACAACACG ATTTTTTCAA ATCAGCCATG CCTGAAGGAT ATGTTCAAGA GCGTACAATC
     2821 AGCTTCAAGG ACGATGGCAC CTATAAAACG CGTGCGGAAG TGAAATTTGA AGGCGACACA
     2881 TTAGTAAACC GTATCGAACT GAAAGGTATC GACTTCAAAG AAGACGGCAA CATTTTAGGC
     2941 CATAAGCTGG AATATAACTT TAATTCTCAT AACGTGTATA TTACGGCCGA TAAACAGAAA
     3001 AACGGTATCA AGGCAAATTT CAAAATTCGC CATAACGTGG AAGACGGCAG CGTTCAATTA
     3061 GCGGATCATT ATCAACAAAA CACGCCGATT GGTGACGGGC CTGTACTGTT ACCTGACAAC
     3121 CACTACCTGA GCACCCAGTC AGCACTGAGC AAAGATCCGA ACGAAAAACG CGATCACATG
     3181 GTTCTGTTAG AATTCGTGAC CGCTGCAGGC ATTACTCACG GAATGGACGA GCTGTACAAG
     3241 AAAGGCGGTT CAAAGAAAAG GAAGAAAAGA GGCGGTAGCA AAAAGCGTAA GAAGCGTGGC
     3301 GGTAGCAAAA AGCGTAAGAA GCGTTAATAA CTCGAGTCTG GTAAAGAAAC CGCTGCTGCG
     3361 AAATTTGAAC GCCAGCACAT GGACTCGTCT ACTAGCGCAG CTTAATTAAC CTAGGCTGCT
     3421 GCCACCGCTG AGCAATAACT AGCATAACCC CTTGGGGCCT CTAAACGGGT CTTGAGGGGT
     3481 TTTTTGCTGA AAGGAGGAAC TATATCCGGA TTGGCGAATG GGACGCGCCC TGTAGCGGCG
     3541 CATTAAGCGC GGCGGGTGTG GTGGTTACGC GCAGCGTGAC CGCTACACTT GCCAGCGCCC
     3601 TAGCGCCCGC TCCTTTCGCT TTCTTCCCTT CCTTTCTCGC CACGTTCGCC GGCTTTCCCC
     3661 GTCAAGCTCT AAATCGGGGG CTCCCTTTAG GGTTCCGATT TAGTGCTTTA CGGCACCTCG
     3721 ACCCCAAAAA ACTTGATTAG GGTGATGGTT CACGTAGTGG GCCATCGCCC TGATAGACGG
     3781 TTTTTCGCCC TTTGACGTTG GAGTCCACGT TCTTTAATAG TGGACTCTTG TTCCAAACTG
     3841 GAACAACACT CAACCCTATC TCGGTCTATT CTTTTGATTT ATAAGGGATT TTGCCGATTT
     3901 CGGCCTATTG GTTAAAAAAT GAGCTGATTT AACAAAAATT TAACGCGAAT TTTAACAAAA
     3961 TATTAACGTT TACAATTTCT GGCGGCACGA TGGCATGAGA TTATCAAAAA GGATCTTCAC
     4021 CTAGATCCTT TTAAATTAAA AATGAAGTTT TAAATCAATC TAAAGTATAT ATGAGTAAAC
     4081 TTGGTCTGAC AGTTACCAAT GCTTAATCAG TGAGGCACCT ATCTCAGCGA TCTGTCTATT
     4141 TCGTTCATCC ATAGTTGCCT GACTCCCCGT CGTGTAGATA ACTACGATAC GGGAGGGCTT
     4201 ACCATCTGGC CCCAGTGCTG CAATGATACC GCGAGACCCA CGCTCACCGG CTCCAGATTT
     4261 ATCAGCAATA AACCAGCCAG CCGGAAGGGC CGAGCGCAGA AGTGGTCCTG CAACTTTATC
     4321 CGCCTCCATC CAGTCTATTA ATTGTTGCCG GGAAGCTAGA GTAAGTAGTT CGCCAGTTAA
     4381 TAGTTTGCGC AACGTTGTTG CCATTGCTAC AGGCATCGTG GTGTCACGCT CGTCGTTTGG
     4441 TATGGCTTCA TTCAGCTCCG GTTCCCAACG ATCAAGGCGA GTTACATGAT CCCCCATGTT
     4501 GTGCAAAAAA GCGGTTAGCT CCTTCGGTCC TCCGATCGTT GTCAGAAGTA AGTTGGCCGC
     4561 AGTGTTATCA CTCATGGTTA TGGCAGCACT GCATAATTCT CTTACTGTCA TGCCATCCGT
     4621 AAGATGCTTT TCTGTGACTG GTGAGTACTC AACCAAGTCA TTCTGAGAAT AGTGTATGCG
     4681 GCGACCGAGT TGCTCTTGCC CGGCGTCAAT ACGGGATAAT ACCGCGCCAC ATAGCAGAAC
     4741 TTTAAAAGTG CTCATCATTG GAAAACGTTC TTCGGGGCGA AAACTCTCAA GGATCTTACC
     4801 GCTGTTGAGA TCCAGTTCGA TGTAACCCAC TCGTGCACCC AACTGATCTT CAGCATCTTT
     4861 TACTTTCACC AGCGTTTCTG GGTGAGCAAA AACAGGAAGG CAAAATGCCG CAAAAAAGGG
     4921 AATAAGGGCG ACACGGAAAT GTTGAATACT CATACTCTTC CTTTTTCAAT CATGATTGAA
     4981 GCATTTATCA GGGTTATTGT CTCATGAGCG GATACATATT TGAATGTATT TAGAAAAATA
     5041 AACAAATAGG TCATGACCAA AATCCCTTAA CGTGAGTTTT CGTTCCACTG AGCGTCAGAC
     5101 CCCGTAGAAA AGATCAAAGG ATCTTCTTGA GATCCTTTTT TTCTGCGCGT AATCTGCTGC
     5161 TTGCAAACAA AAAAACCACC GCTACCAGCG GTGGTTTGTT TGCCGGATCA AGAGCTACCA
     5221 ACTCTTTTTC CGAAGGTAAC TGGCTTCAGC AGAGCGCAGA TACCAAATAC TGTCCTTCTA
     5281 GTGTAGCCGT AGTTAGGCCA CCACTTCAAG AACTCTGTAG CACCGCCTAC ATACCTCGCT
     5341 CTGCTAATCC TGTTACCAGT GGCTGCTGCC AGTGGCGATA AGTCGTGTCT TACCGGGTTG
     5401 GACTCAAGAC GATAGTTACC GGATAAGGCG CAGCGGTCGG GCTGAACGGG GGGTTCGTGC
     5461 ACACAGCCCA GCTTGGAGCG AACGACCTAC ACCGAACTGA GATACCTACA GCGTGAGCTA
     5521 TGAGAAAGCG CCACGCTTCC CGAAGGGAGA AAGGCGGACA GGTATCCGGT AAGCGGCAGG
     5581 GTCGGAACAG GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA TCTTTATAGT
     5641 CCTGTCGGGT TTCGCCACCT CTGACTTGAG CGTCGATTTT TGTGATGCTC GTCAGGGGGG
     5701 CGGAGCCTAT GGAAAAACGC CAGCAACGCG GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG
     5761 CCTTTTGCTC ACATGTTCTT TCCTGCGTTA TCCCCTGATT CTGTGGATAA CCGTATTACC
     5821 GCCTTTGAGT GAGCTGATAC CGCTCGCCGC AGCCGAACGA CCGAGCGCAG CGAGTCAGTG
     5881 AGCGAGGAAG CGGAAGAGCG CCTGATGCGG TATTTTCTCC TTACGCATCT GTGCGGTATT
     5941 TCACACCGCA TATATGGTGC ACTCTCAGTA C
//