LOCUS       SMT503_S77A             3894 bp ds-DNA     circular     28-AUG-2024
DEFINITION  .
COMMENT     This plasmid is an derived from SMT213-SNAP-ccdb-eGFP created by
            Daria Passow. It constains the lipase 3QZU and the S77A catalytic
            serine mutation. Chloramphenicol resistance. Sequence files from
            Golden Gate cloning of the lipase Sequence files from Gibson
            cloning of SNAP and CLIP Sequence files from Gibson cloning of the
            ccdb gene Original Sequencing Files
FEATURES             Location/Qualifiers
     misc_feature    180..205
                     /label="CamR REV primer"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     stem_loop       180..285
                     /label="T0 terminator"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     cds             complement(298..957)
                     /label="Chloramphenicol resistance"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    complement(958..1063)
                     /label="Unnanotated Promoter (P1004)"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer          1042..1063
                     /label="SMT000 NAQ_seq1"
                     /note="sequence: ACCTCTTACGTGCCCGATCAAC"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          1042..1063
                     /label="SMT2XX_IVTT_2bio_Fwd"
                     /note="sequence: acctcttacgtgcccgatcaac"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     primer          1042..1063
                     /label="SMT2XX_IVTT_Fwd"
                     /note="sequence: acctcttacgtgcccgatcaac"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          1042..1063
                     /label="SMT000_IVTT_fwd"
                     /note="sequence: ACCTCTTACGTGCCCGATCAAC"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    complement(1042..1063)
                     /label="CamR FOR primer"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          1068..1087
                     /label="SMT2XX_nestedFwd"
                     /note="sequence: gtGCCACCTGACGTCTAAGA"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    complement(1070..1091)
                     /label="pSB1C3 anneal REV"
                     /ApEinfo_revcolor="#c6c9d1"
                     /ApEinfo_fwdcolor="#c6c9d1"
     primer_binding  1071..1090
                     /label="Verification forward (VF2) primer binding site"
                     /ApEinfo_revcolor="#f8d3a9"
                     /ApEinfo_fwdcolor="#f8d3a9"
     primer          1078..1105
                     /label="SMT2XX_gib_5ins_fwd"
                     /note="sequence: ACGTCTAAGAAACCgaataatacgactc"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    1078..1137
                     /label="5' Forward Homology Arm for Gibson"
                     /ApEinfo_revcolor="#b7e6d7"
                     /ApEinfo_fwdcolor="#b7e6d7"
     misc_feature    1095..1111
                     /label="T7 promoter"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     primer          complement(1105..1137)
                     /label="SMT2XX_5lin_rev"
                     /note="sequence: ATGTATATCTCCTTatacttaagccctatagtg"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1124..1130
                     /label="RBS?"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1134..1137
                     /label="AGGA"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1138..1140
                     /label="Start Codon"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    1138..1679
                     /label="SNAP_legacy"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     CDS             1138..1679
                     /label="Translation 1138-1679"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHEIKLLGKGTSAADAVEVPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLKVVKFGEVISYQQLAALAGNPAATAAVKTALSGNPVPILIPCHRVVSSSGAVGGYEGGLAVKEWLLAHEGHRLGKPG"
     primer          complement(1174..1192)
                     /label="GGseq_SNAP_Rev"
                     /note="sequence: gtttgcccagcgggctatc"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          1606..1625
                     /label="GGseq_SNAP_Fwd"
                     /note="sequence: ggttatgaaggtggtctggc"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     CDS             1678..2286
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="LGGGGSGGGGSGAEHNPVVMVHGIGGASFNFAGIKSYLVSQGWSQNDLYAVDFWDKTGTNYNNGPVLSRFVQKVLDETGAKKVDIVAHAMGGANTLYYIKNLDGGNKVANVVTLGGANRLTTGDALPGTDPNQKILYTSIYSSADDIVMNCLSRLDGARNVQIHGVGHMGLLYSSQVNSLIKEGLNGGGQNTNGGGSGGGGSG"
     misc_feature    1714..2256
                     /label="3QZU_S77A - B. subtilis lipase, catalytic mutant"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     CDS             2285..2286
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation=""
     misc_feature    2287..2997
                     /label="eGFP (first MV deleted)"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             2287..2997
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer          complement(2977..3035)
                     /label="primDRW-023_3ins_eGFP_Rev"
                     /note="sequence: gtagcGATCTACACTAGCACTATCAGCGGGAATCATTATTTGTACAGCTCGTCCATGCC"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    2998..3003
                     /label="Double Stop Codon"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     CDS             2998..3003
                     /label="Translation 2998-3003"
                     /translation="**"
     misc_feature    2998..3057
                     /label="3' Reverse Homology Arm for Gibson"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    3004..3007
                     /label="TTCC"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          3004..3024
                     /label="SMT2XX_3lin_fwd"
                     /note="sequence: TTCCCGCTGATAGTGCTAGTG"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             3005..3007
                     /label="S*"
                     /ApEinfo_revcolor="#993366"
                     /ApEinfo_fwdcolor="#993366"
     misc_feature    3008..3032
                     /label="pSB1C3 anneal REV"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     barcode         3008..3032
                     /label="Help:Barcodes"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          complement(3031..3048)
                     /label="SMT2XX_gib_3ins_rev"
                     /note="sequence: gcggccgctactagtagc"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     dna             3033..3053
                     /label="BioBrick suffix"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(3055..3072)
                     /label="SMT2XX_seq_Rev"
                     /note="sequence: cttgcccttttttgccgg"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     stem_loop       3058..3125
                     /label="E. coli his operon terminator"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     stem_loop       3062..3096
                     /label="Stem loop"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(3067..3082)
                     /label="SMT000 Gibs Ins Rnd2 Rev"
                     /note="sequence: gtggtgacaccttgcc"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    complement(3110..3134)
                     /label="pSB1C3 anneal REV"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     primer          complement(3115..3134)
                     /label="SMT2XX_nestedRev"
                     /note="sequence: gaagcctgcataacgcgaag"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          complement(3160..3175)
                     /label="SMT000_bead_amp_50_rev"
                     /note="sequence: gccgcagccgaacgac"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     primer          complement(3188..3211)
                     /label="SMT000_IVTT_rev"
                     /note="sequence: ACCGTATTACCGCCTTTGAGTGAG"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     primer          complement(3188..3211)
                     /label="SMT2XX_IVTT_2bio_Rev"
                     /note="sequence: accgtattaccgcctttgagtgag"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer          complement(3188..3211)
                     /label="SMT2XX_IVTT_amino_Rev"
                     /note="sequence: accgtattaccgcctttgagtgag"
                     /ApEinfo_revcolor="#75c6a9"
                     /ApEinfo_fwdcolor="#75c6a9"
     primer_binding  complement(3189..3208)
                     /label="Verification reverse (VR) primer binding site"
                     /ApEinfo_revcolor="#d59687"
                     /ApEinfo_fwdcolor="#d59687"
     primer          complement(3258..3275)
                     /label="SMT000_bead_amp_150_rev"
                     /note="sequence: ctggccttttgctggcct"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc            complement(3293..13)
                     /label="rep (pMB1)"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc            3308..3308
                     /label="ORI"
                     /ApEinfo_revcolor="#9eafd2"
                     /ApEinfo_fwdcolor="#9eafd2"
     primer          complement(3358..3375)
                     /label="SMT000_bead_amp_250_rev"
                     /note="sequence: cgggtttcgccacctctg"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     primer          complement(3600..3625)
                     /label="SMT000_bead_amp_500_rev"
                     /note="sequence: ggttggactcaagacgatagttaccg"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
      121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ctcgaggctt
      181 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
      241 agttctgagg tcattactgg atctatcaac aggagtccaa gcgagctcga tatcaaatta
      301 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
      361 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
      421 ttgcgtataa tatttgccca tggtgaaaac gggggcgaag aagttgtcca tattggccac
      481 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
      541 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
      601 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
      661 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
      721 gtctttcatt gccatacgaa attccggatg agcattcatc aggcgggcaa gaatgtgaat
      781 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
      841 cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc
      901 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
      961 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     1021 ttcattatgg tgaaagttgg aacctcttac gtgcccgatc aactcgagtG CCACCTGACG
     1081 TCTAAGAAAC Cgaataatac gactcactat agggcttaag tatAAGGAGA TATACATATG
     1141 gacaaagatt gcgaaatgaa acgtaccacc ctggatagcc cgctgggcaa actggaactg
     1201 agcggctgcg aacagggcct gcatgaaatt aaactgctgg gtaaaggcac cagcgcggcc
     1261 gatgcggttg aagttccggc cccggccgcc gtgctgggtg gtccggaacc gctgatgcag
     1321 gcgaccgcgt ggctgaacgc gtattttcat cagccggaag cgattgaaga atttccggtt
     1381 ccggcgctgc atcatccggt gtttcagcag gagagcttta cccgtcaggt gctgtggaaa
     1441 ctgctgaaag tggttaaatt tggcgaagtg attagctatc agcagctggc ggccctggcg
     1501 ggtaatccgg cggccaccgc cgccgttaaa accgcgctga gcggtaaccc ggtgccgatt
     1561 ctgattccgt gccatcgtgt ggttagctct agcggtgcgg ttggcggtta tgaaggtggt
     1621 ctggcggtga aagagtggct gctggcccat gaaggtcatc gtctgggtaa accgggtctG
     1681 GGAGGTGGAG GGTCTGGGGG AGGAGGCTCA GGCGCGGAAC ATAACCCGGT GGTGATGGTG
     1741 CATGGCATTG GCGGTGCGAG CTTTAACTTT GCGGGCATTA AATCTTATCT GGTGAGCCAG
     1801 GGCTGGAGCC AGAACGATCT GTATGCGGTG GACTTTTGGG ATAAAACCGG TACCAACTAT
     1861 AACAACGGCC CGGTGCTGAG CCGTTTTGTG CAGAAAGTGC TGGACGAAAC CGGCGCGAAG
     1921 AAAGTGGATA TCGTGGCGCA TGCGATGGGT GGCGCGAACA CCCTGTATTA TATTAAGAAC
     1981 CTGGACGGCG GCAACAAAGT GGCGAACGTG GTGACCCTGG GTGGTGCAAA CCGCTTAACC
     2041 ACAGGTGATG CGCTGCCGGG TACCGATCCG AACCAGAAAA TTCTGTATAC CAGCATTTAC
     2101 AGCAGCGCGG ATGATATTGT GATGAACTGT CTGAGCCGCC TGGATGGCGC GCGCAACGTT
     2161 CAGATTCATG GCGTGGGTCA TATGGGCCTG CTGTATAGCA GCCAGGTGAA CAGCCTGATT
     2221 AAAGAAGGCC TGAACGGTGG CGGCCAGAAC ACCAACGGTG GCGGTTCTGG CGGTGGCGGT
     2281 AGTGGCAGCA AGGGCGAGGA GCTGTTCACC GGGGTGGTGC CCATCCTGGT CGAGCTGGAC
     2341 GGCGACGTAA ACGGCCACAA GTTCAGCGTG TCCGGCGAGG GCGAGGGCGA TGCCACCTAC
     2401 GGCAAGCTGA CCCTGAAGTT CATCTGCACC ACCGGCAAGC TGCCCGTGCC CTGGCCCACC
     2461 CTCGTGACCA CCCTGACCTA CGGCGTGCAG TGCTTCAGCC GCTACCCCGA CCACATGAAG
     2521 CAGCACGACT TCTTCAAGTC CGCCATGCCC GAAGGCTACG TCCAGGAGCG CACCATCTTC
     2581 TTCAAGGACG ACGGCAACTA CAAGACCCGC GCCGAGGTGA AGTTCGAGGG CGACACCCTG
     2641 GTGAACCGCA TCGAGCTGAA GGGCATCGAC TTCAAGGAGG ACGGCAACAT CCTGGGGCAC
     2701 AAGCTGGAGT ACAACTACAA CAGCCACAAC GTCTATATCA TGGCCGACAA GCAGAAGAAC
     2761 GGCATCAAGG TGAACTTCAA GATCCGCCAC AACATCGAGG ACGGCAGCGT GCAGCTCGCC
     2821 GACCACTACC AGCAGAACAC CCCCATCGGC GACGGCCCCG TGCTGCTGCC CGACAACCAC
     2881 TACCTGAGCA CCCAGTCCGC CCTGAGCAAA GACCCCAACG AGAAGCGCGA TCACATGGTC
     2941 CTGCTGGAGT TCGTGACCGC CGCCGGGATC ACTCTCGGCA TGGACGAGCT GTACAAATAA
     3001 TGATTCCCGC TGATAGTGCT AGTGTAGATC gctactagta gcggccgctg cagtccggca
     3061 aaaaagggca aggtgtcacc accctgccct ttttctttaa aaccgaaaag attacttcgc
     3121 gttatgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3181 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3241 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3301 gcgtttttcc acaggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3361 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3421 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3481 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3541 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3601 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3661 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3721 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3781 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3841 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaa
//