LOCUS       #652_pET-28a(+)-6xHis-L 5970 bp ds-DNA     circular     15-MAR-2024
DEFINITION  .
KEYWORDS    "Plasmid name:pET-28a(+)-6xHis-LSSmCherry1 (-13)" "Gene insert
            **:6xHis-LSSmCherry1 (-13)" "Alt. gene insert name" "Relevant
            mutations" "Prepared by **:So Yeon Ahn" "Cloning method **:Hifi
            assembly" "Resistance **:Kanamycin (Kan)" "Backbone **:pET-28a(+)"
            "Purification tag **:6xHis" "N-terminal tag:6xHis" "C-terminal tag"
            "Secretion signal (for yeast only)" "Glycerol stock numbers:652,
            653" "Sequencing file and other attachments" "Other notes"
COMMENT     Imported using the Genbank importer. File name: 6xHis-mBlueberry.gb
            6xHis-mBlueberry in pET-28a(+) Resistance markers: Kanamycin This
            file was created using tools provided by Twist Bioscience
FEATURES             Location/Qualifiers
     misc_feature    1..60
                     /label="Affirmative pET-28a(+), vector ordered from TWIST"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature_   61..78
                     /label="T7_pro"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     misc_feature_   79..105
                     /label="lacO"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature_   119..140
                     /label="RBS_3"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature_   120..139
                     /label="RBS_4"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature_   124..139
                     /label="T7_transl_en_RBS"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     CDS             148..888
                     /label="Translation 148-888"
                     /translation="MGHHHHHHGGVSKGEEDNMTIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFNWERVMNFEDGGVVTVTQDSSLQDGEFIYKVELRGTNFPSDGPVMQCRTMGLEASTERMYPEDGALKGESKERLKLKDGGHYDAEVKTTYKAKEPVQLPGAYNVDIKLDILSHNEDYTIVEQYERSEGRHSTGGMDELYK**"
     misc_feature_   152..888
                     /label="LSSmCherry1 (-13)"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     misc_feature    154..171
                     /label="His6x"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          546..567
                     /label="[ASY36] LSSmCherry_Mut1_fwd"
                     /note="sequence: ctacaaggtggagctgcgcggc"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(546..567)
                     /label="[ASY 37] LSSmCherry_Mut1_rev"
                     /note="sequence: gccgcgcagctccaccttgtag"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    556..558
                     /label="K137E"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="AAG->GAG"
     primer          732..753
                     /label="[ASY38] LSSmCherry_Mut2_fwd"
                     /note="sequence: caaggccaaggagcccgtgcag"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     primer          complement(732..753)
                     /label="[ASY39] LSSmCherry_Mut2_rev"
                     /note="sequence: ctgcacgggctccttggccttg"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature    742..744
                     /label="K199E"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
                     /note="AAG->GAG"
     misc_feature_   889..5970
                     /label="vector_backbone-737-5970"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature_   923..1050
                     /label="T7"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature_   1080..1385
                     /label="f1"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     misc_feature_   1610..2424
                     /label="kan"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     primer          complement(2407..2434)
                     /label="Mut_Kan_rev (SYA 11)"
                     /note="sequence: AGGGGTGTTATGAGCCATATTCAACGGG"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer          2415..2450
                     /label="Kan-LacI_fwd (SYA 1)"
                     /note="sequence: ATATGGCTCATAACACCCCTTGTATTACTGTTTATG"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature_   2563..3134
                     /label="colE1_high_copy"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature_   3562..3752
                     /label="ROP"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature_   4528..4548
                     /label="CAP_binding"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature_   4562..5652
                     /label="lacI"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     misc_feature_   5645..5721
                     /label="lacIQ_promoter"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
ORIGIN
        1 CCGGTGATGC CGGCCACGAT GCGTCCGGCG TAGAGGATCG AGATCTCGAT CCCGCGAAAT
       61 TAATACGACT CACTATAGGG GAATTGTGAG CGGATAACAA TTCCCCTCTA GAAATAATTT
      121 TGTTTAACTT TAAGAAGGAG ATATACCATG GGCCATCATC ATCATCATCA TGGCGGCgtg
      181 agcaagggcg aggaggataa catgaccatc atcaaggagt tcatgcgctt caaggtgcac
      241 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
      301 tacgagggca cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc
      361 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
      421 gacatccccg actacttgaa gctgtccttc cccgagggct tcaattggga gcgcgtgatg
      481 aactttgagg acggcggtgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
      541 ttcatctaca aggtggagct gcgcggcacc aacttcccct ctgacggccc cgtaatgcag
      601 tgtcgtacca tgggcttgga ggccagtact gagcggatgt accccgagga cggcgccctg
      661 aagggcgaga gtaaggagag gctgaagctg aaggacggcg gccactacga cgctgaggtc
      721 aagaccacct acaaggccaa ggagcccgtg cagctgcccg gcgcctacaa cgtcgacatc
      781 aagttggaca tcctttccca caacgaggac tacaccatcg tggaacagta cgaacgctcc
      841 gagggccgcc actccaccgg cggcatggac gagctgtaca agTAATAACT CGAGCACCAC
      901 CACCACCACC ACTGAGATCC GGCTGCTAAC AAAGCCCGAA AGGAAGCTGA GTTGGCTGCT
      961 GCCACCGCTG AGCAATAACT AGCATAACCC CTTGGGGCCT CTAAACGGGT CTTGAGGGGT
     1021 TTTTTGCTGA AAGGAGGAAC TATATCCGGA TTGGCGAATG GGACGCGCCC TGTAGCGGCG
     1081 CATTAAGCGC GGCGGGTGTG GTGGTTACGC GCAGCGTGAC CGCTACACTT GCCAGCGCCC
     1141 TAGCGCCCGC TCCTTTCGCT TTCTTCCCTT CCTTTCTCGC CACGTTCGCC GGCTTTCCCC
     1201 GTCAAGCTCT AAATCGGGGG CTCCCTTTAG GGTTCCGATT TAGTGCTTTA CGGCACCTCG
     1261 ACCCCAAAAA ACTTGATTAG GGTGATGGTT CACGTAGTGG GCCATCGCCC TGATAGACGG
     1321 TTTTTCGCCC TTTGACGTTG GAGTCCACGT TCTTTAATAG TGGACTCTTG TTCCAAACTG
     1381 GAACAACACT CAACCCTATC TCGGTCTATT CTTTTGATTT ATAAGGGATT TTGCCGATTT
     1441 CGGCCTATTG GTTAAAAAAT GAGCTGATTT AACAAAAATT TAACGCGAAT TTTAACAAAA
     1501 TATTAACGCT TACAATTTAG GTGGCACTTT TCGGGGAAAT GTGCGCGGAA CCCCTATTTG
     1561 TTTATTTTTC TAAATACATT CAAATATGTA TCCGCTCATG AATTAATTCT TAGAAAAACT
     1621 CATCGAGCAT CAAATGAAAC TGCAATTTAT TCATATCAGG ATTATCAATA CCATATTTTT
     1681 GAAAAAGCCG TTTCTGTAAT GAAGGAGAAA ACTCACCGAG GCAGTTCCAT AGGATGGCAA
     1741 GATCCTGGTA TCGGTCTGCG ATTCCGACTC GTCCAACATC AATACAACCT ATTAATTTCC
     1801 CCTCGTCAAA AATAAGGTTA TCAAGTGAGA AATCACCATG AGTGACGACT GAATCCGGTG
     1861 AGAATGGCAA AAGTTTATGC ATTTCTTTCC AGACTTGTTC AACAGGCCAG CCATTACGCT
     1921 CGTCATCAAA ATCACTCGCA TCAACCAAAC CGTTATTCAT TCGTGATTGC GCCTGAGCGA
     1981 GACGAAATAC GCGATCGCTG TTAAAAGGAC AATTACAAAC AGGAATCGAA TGCAACCGGC
     2041 GCAGGAACAC TGCCAGCGCA TCAACAATAT TTTCACCTGA ATCAGGATAT TCTTCTAATA
     2101 CCTGGAATGC TGTTTTCCCG GGGATCGCAG TGGTGAGTAA CCATGCATCA TCAGGAGTAC
     2161 GGATAAAATG CTTGATGGTC GGAAGAGGCA TAAATTCCGT CAGCCAGTTT AGTCTGACCA
     2221 TCTCATCTGT AACATCATTG GCAACGCTAC CTTTGCCATG TTTCAGAAAC AACTCTGGCG
     2281 CATCGGGCTT CCCATACAAT CGATAGATTG TCGCACCTGA TTGCCCGACA TTATCGCGAG
     2341 CCCATTTATA CCCATATAAA TCAGCATCCA TGTTGGAATT TAATCGCGGC CTAGAGCAAG
     2401 ACGTTTCCCG TTGAATATGG CTCATAACAC CCCTTGTATT ACTGTTTATG TAAGCAGACA
     2461 GTTTTATTGT TCATGACCAA AATCCCTTAA CGTGAGTTTT CGTTCCACTG AGCGTCAGAC
     2521 CCCGTAGAAA AGATCAAAGG ATCTTCTTGA GATCCTTTTT TTCTGCGCGT AATCTGCTGC
     2581 TTGCAAACAA AAAAACCACC GCTACCAGCG GTGGTTTGTT TGCCGGATCA AGAGCTACCA
     2641 ACTCTTTTTC CGAAGGTAAC TGGCTTCAGC AGAGCGCAGA TACCAAATAC TGTCCTTCTA
     2701 GTGTAGCCGT AGTTAGGCCA CCACTTCAAG AACTCTGTAG CACCGCCTAC ATACCTCGCT
     2761 CTGCTAATCC TGTTACCAGT GGCTGCTGCC AGTGGCGATA AGTCGTGTCT TACCGGGTTG
     2821 GACTCAAGAC GATAGTTACC GGATAAGGCG CAGCGGTCGG GCTGAACGGG GGGTTCGTGC
     2881 ACACAGCCCA GCTTGGAGCG AACGACCTAC ACCGAACTGA GATACCTACA GCGTGAGCTA
     2941 TGAGAAAGCG CCACGCTTCC CGAAGGGAGA AAGGCGGACA GGTATCCGGT AAGCGGCAGG
     3001 GTCGGAACAG GAGAGCGCAC GAGGGAGCTT CCAGGGGGAA ACGCCTGGTA TCTTTATAGT
     3061 CCTGTCGGGT TTCGCCACCT CTGACTTGAG CGTCGATTTT TGTGATGCTC GTCAGGGGGG
     3121 CGGAGCCTAT GGAAAAACGC CAGCAACGCG GCCTTTTTAC GGTTCCTGGC CTTTTGCTGG
     3181 CCTTTTGCTC ACATGTTCTT TCCTGCGTTA TCCCCTGATT CTGTGGATAA CCGTATTACC
     3241 GCCTTTGAGT GAGCTGATAC CGCTCGCCGC AGCCGAACGA CCGAGCGCAG CGAGTCAGTG
     3301 AGCGAGGAAG CGGAAGAGCG CCTGATGCGG TATTTTCTCC TTACGCATCT GTGCGGTATT
     3361 TCACACCGCA ATGGTGCACT CTCAGTACAA TCTGCTCTGA TGCCGCATAG TTAAGCCAGT
     3421 ATACACTCCG CTATCGCTAC GTGACTGGGT CATGGCTGCG CCCCGACACC CGCCAACACC
     3481 CGCTGACGCG CCCTGACGGG CTTGTCTGCT CCCGGCATCC GCTTACAGAC AAGCTGTGAC
     3541 CGTCTCCGGG AGCTGCATGT GTCAGAGGTT TTCACCGTCA TCACCGAAAC GCGCGAGGCA
     3601 GCTGCGGTAA AGCTCATCAG CGTGGTCGTG AAGCGATTCA CAGATGTCTG CCTGTTCATC
     3661 CGCGTCCAGC TCGTTGAGTT TCTCCAGAAG CGTTAATGTC TGGCTTCTGA TAAAGCGGGC
     3721 CATGTTAAGG GCGGTTTTTT CCTGTTTGGT CACTGATGCC TCCGTGTAAG GGGGATTTCT
     3781 GTTCATGGGG GTAATGATAC CGATGAAACG AGAGAGGATG CTCACGATAC GGGTTACTGA
     3841 TGATGAACAT GCCCGGTTAC TGGAACGTTG TGAGGGTAAA CAACTGGCGG TATGGATGCG
     3901 GCGGGACCAG AGAAAAATCA CTCAGGGTCA ATGCCAGCGC TTCGTTAATA CAGATGTAGG
     3961 TGTTCCACAG GGTAGCCAGC AGCATCCTGC GATGCAGATC CGGAACATAA TGGTGCAGGG
     4021 CGCTGACTTC CGCGTTTCCA GACTTTACGA AACACGGAAA CCGAAGACCA TTCATGTTGT
     4081 TGCTCAGGTC GCAGACGTTT TGCAGCAGCA GTCGCTTCAC GTTCGCTCGC GTATCGGTGA
     4141 TTCATTCTGC TAACCAGTAA GGCAACCCCG CCAGCCTAGC CGGGTCCTCA ACGACAGGAG
     4201 CACGATCATG CGCACCCGTG GGGCCGCCAT GCCGGCGATA ATGGCCTGCT TCTCGCCGAA
     4261 ACGTTTGGTG GCGGGACCAG TGACGAAGGC TTGAGCGAGG GCGTGCAAGA TTCCGAATAC
     4321 CGCAAGCGAC AGGCCGATCA TCGTCGCGCT CCAGCGAAAG CGGTCCTCGC CGAAAATGAC
     4381 CCAGAGCGCT GCCGGCACCT GTCCTACGAG TTGCATGATA AAGAAGACAG TCATAAGTGC
     4441 GGCGACGATA GTCATGCCCC GCGCCCACCG GAAGGAGCTG ACTGGGTTGA AGGCTCTCAA
     4501 GGGCATCGGT CGAGATCCCG GTGCCTAATG AGTGAGCTAA CTTACATTAA TTGCGTTGCG
     4561 CTCACTGCCC GCTTTCCAGT CGGGAAACCT GTCGTGCCAG CTGCATTAAT GAATCGGCCA
     4621 ACGCGCGGGG AGAGGCGGTT TGCGTATTGG GCGCCAGGGT GGTTTTTCTT TTCACCAGTG
     4681 AGACGGGCAA CAGCTGATTG CCCTTCACCG CCTGGCCCTG AGAGAGTTGC AGCAAGCGGT
     4741 CCACGCTGGT TTGCCCCAGC AGGCGAAAAT CCTGTTTGAT GGTGGTTAAC GGCGGGATAT
     4801 AACATGAGCT GTCTTCGGTA TCGTCGTATC CCACTACCGA GATATCCGCA CCAACGCGCA
     4861 GCCCGGACTC GGTAATGGCG CGCATTGCGC CCAGCGCCAT CTGATCGTTG GCAACCAGCA
     4921 TCGCAGTGGG AACGATGCCC TCATTCAGCA TTTGCATGGT TTGTTGAAAA CCGGACATGG
     4981 CACTCCAGTC GCCTTCCCGT TCCGCTATCG GCTGAATTTG ATTGCGAGTG AGATATTTAT
     5041 GCCAGCCAGC CAGACGCAGA CGCGCCGAGA CAGAACTTAA TGGGCCCGCT AACAGCGCGA
     5101 TTTGCTGGTG ACCCAATGCG ACCAGATGCT CCACGCCCAG TCGCGTACCG TCTTCATGGG
     5161 AGAAAATAAT ACTGTTGATG GGTGTCTGGT CAGAGACATC AAGAAATAAC GCCGGAACAT
     5221 TAGTGCAGGC AGCTTCCACA GCAATGGCAT CCTGGTCATC CAGCGGATAG TTAATGATCA
     5281 GCCCACTGAC GCGTTGCGCG AGAAGATTGT GCACCGCCGC TTTACAGGCT TCGACGCCGC
     5341 TTCGTTCTAC CATCGACACC ACCACGCTGG CACCCAGTTG ATCGGCGCGA GATTTAATCG
     5401 CCGCGACAAT TTGCGACGGC GCGTGCAGGG CCAGACTGGA GGTGGCAACG CCAATCAGCA
     5461 ACGACTGTTT GCCCGCCAGT TGTTGTGCCA CGCGGTTGGG AATGTAATTC AGCTCCGCCA
     5521 TCGCCGCTTC CACTTTTTCC CGCGTTTTCG CAGAAACGTG GCTGGCCTGG TTCACCACGC
     5581 GGGAAACGGT CTGATAAGAG ACACCGGCAT ACTCTGCGAC ATCGTATAAC GTTACTGGTT
     5641 TCACATTCAC CACCCTGAAT TGACTCTCTT CCGGGCGCTA TCATGCCATA CCGCGAAAGG
     5701 TTTTGCGCCA TTCGATGGTG TCCGGGATCT CGACGCTCTC CCTTATGCGA CTCCTGCATT
     5761 AGGAAGCAGC CCAGTAGTAG GTTGAGGCCG TTGAGCACCG CCGCCGCAAG GAATGGTGCA
     5821 TGCAAGGAGA TGGCGCCCAA CAGTCCCCCG GCCACGGGGC CTGCCACCAT ACCCACGCCG
     5881 AAACAAGCGC TCATGAGCCC GAAGTGGCGA GCCCGATCTT CCCCATCGGT GATGTCGGCG
     5941 ATATAGGCGC CAGCAACCGC ACCTGTGGCG
//