LOCUS       pYSD021                 1926 bp ds-DNA     circular     12-NOV-2023
DEFINITION  .
ACCESSION   <unknown id>
VERSION     <unknown id>
COMMENT     Encodes an engineered S. cerevisiae MFalpha1 pre-pro- signal
            peptide (MFapp8) as a Type 3a’ part to be used in the yeast
            secretion/display (YSD) extension of the MoClo yeast toolkit (YTK)
            Vector backbone is the Yeast MoClo toolkit entry vector pYTK001
            MFapp8 reference: Rakestraw, J.A., Sazinsky, S.L., Piatesi, A.,
            Antipov, E., and Wittrup, K.D. (2009). Directed evolution of a
            secretory leader for the improved expression of heterologous
            proteins and full-length antibodies in Saccharomyces cerevisiae.
            Biotechnol Bioeng 103(6), 1192-1201. doi: 10.1002/bit.22338.
FEATURES             Location/Qualifiers
     misc_feature    11..277
                     /label="Engineered Mating Factor alpha 1 Pre- Pro- app8 Signal Peptide"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     CDS             11..277
                     /label="MFalpha pre-pro- app8 engineered variant"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /translation="MRFPSIFTAVLFAASSALAAPANTTTEDETAQIPAEAVIDYSDLEGDFDAAALPLSNSTNNGLSSTNTTIASIAAKEEGVQLDKREADA"
     misc_feature    254..265
                     /label="Kex2p Cleavage Site "
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    272..274
                     /label="E to D comp to WT seq S to A comp to yeast display vector"
                     /ApEinfo_revcolor="#d6b295"
                     /ApEinfo_fwdcolor="#d6b295"
     misc_feature    complement(287..395)
                     /label="CamR Terminator"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     CDS             complement(396..1055)
                     /label="CamR"
                     /ApEinfo_revcolor="#0000ff"
                     /ApEinfo_fwdcolor="#0000ff"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFLKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA*"
     promoter        complement(1056..1160)
                     /label="CamR Promoter"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
     rep_origin      complement(1161..1924)
                     /label="ColE1"
                     /ApEinfo_revcolor="#7f7f7f"
                     /ApEinfo_fwdcolor="#7f7f7f"
                     /ApEinfo_graphicformat="arrow_data {{0 1 2 0 0 -1} {} 0}width 5 offset 0"
ORIGIN
        1 TCGGTCTCAT ATGAGATTCC CATCTATCTT CACCGCTGTT TTGTTCGCTG CTTCTTCTGC
       61 TTTGGCTGCT CCAGCTAACA CCACCACCGA AGACGAAACC GCTCAAATCC CAGCTGAAGC
      121 TGTTATCGAC TACTCTGACT TGGAAGGTGA CTTCGACGCT GCTGCTTTGC CATTGTCTAA
      181 CTCTACCAAC AACGGTTTGT CTTCTACCAA CACCACCATC GCTTCTATCG CTGCTAAGGA
      241 AGAAGGTGTT CAATTGGACA AGAGAGAAGC TgatGCtTGA gaccagacca ataaaaaacg
      301 cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc attactggat
      361 ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg ccactcatcg
      421 cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca aacggcatga
      481 tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata tttgcccatg
      541 gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa actggtgaaa
      601 ctcacccagg gattggctga aacgaaaaac atattctcaa taaacccttt agggaaatag
      661 gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa ctgccggaaa
      721 tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg gaaaacggtg
      781 taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc catacgaaat
      841 tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata aaacttgtgc
      901 ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt ctggttatag
      961 gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca ttgggatata
     1021 tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc tcctgaaaat
     1081 ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg aaagttggaa
     1141 cctcttacgt gcccgatcaa tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     1201 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     1261 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     1321 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     1381 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     1441 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     1501 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     1561 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     1621 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     1681 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     1741 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     1801 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     1861 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     1921 ccgtag
//