LOCUS       sequence-421794-        3820 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the HA-tagged 649 Stalk yeast surface display anchor protein
            as a Type 4a part to be used in the yeast secretion/display (YSD) 
            extension of the MoClo yeast toolkit (YTK).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3820)
  AUTHORS   O'Riordan NM, Juric V, O'Neill SK, Roche AP, Young PW
  TITLE     A Yeast Modular Cloning (MoClo) Toolkit Expansion for Optimization 
            of Heterologous Protein Secretion and Surface Display in 
            Saccharomyces cerevisiae.
  JOURNAL   ACS Synth Biol. 2024 Mar 14. doi: 10.1021/acssynbio.3c00743.
  PUBMED    38483353
REFERENCE   2  (bases 1 to 3820)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol. 2024 Mar 14. doi: 10.1021/acssynbio.3c00743."
COMMENT     Sequence Label: pYSD085
FEATURES             Location/Qualifiers
     source          1..3820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1786..1806)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(1810..1836)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     rep_origin      complement(1981..2569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2061..2080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             2751..3410
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2817..2836)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3481..3515
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             3574..3591
                     /codon_start=1
                     /product="HQHQHQ metal affinity tag"
                     /label=HQ tag
                     /note="The HQ tag binds to nickel and other metal ions much
                     like the 6xHis tag, but may allow elution at lower 
                     concentrations of imidazole (Godat et al., 2008)."
                     /translation="HQHQHQ"
ORIGIN
        1 agaagcagtg gatgatttat cttctgtcga tgtgcttgag gagtgggtag acttactatc
       61 ggactgtgct gtcatagtgg aagtagtgtg cgacaatttc gttgttggcg tagagacaaa
      121 tgaagtggtt gaggggacac ctcctatagt aggagaagtt tcgaaagttt gcgaggttga
      181 tggtgaggta gaagctgtgg agctagatga agtagattcc gaactggttg ttgttgatgt
      241 tgaactaggc gtagatgaca tgacagactg aacctcagaa gattccgagc tgctagtaga
      301 actagaagaa taagctgttg tcgtagacat cactggacct gttgatgtag taatagtagt
      361 ggtggttact acagtacttt gagaatgaga ggtagctggt gttgatacag cagaagttgt
      421 caatgagctg gtggaggaag aactcacgga agatggtgat ggagatggag tgcttgaaga
      481 tgtcgatgtg cttgatgttg tagaggtaga actcttagaa gagacacttg atgaaggtgc
      541 tgttgatgga gcatcttctg ctgatgtcga ttcaaaactt tcagattcgc tagaaattga
      601 ggacgatgta gatgaagatg tcgaggaaga gggataggta atactagatt gagatttcga
      661 cgatatgtga gcgctactac ttgaagttgt aattgttgaa ctagatgtag atgtagtaga
      721 tgaactggct tcaatcgaag tttgtgtggt agagactgac aaggttgcgc tgttagaagt
      781 ggtcgtggca gatgatgaag aagaagaagt tgtcgtttct ccactggaaa tagaagtgga
      841 agtagaagtg ctactagaag atgttgatgt agaacttcca ggtgatacgg aatcggatgg
      901 tgttgcagac tcagaggttt gagatgtact tgaagaagaa gtagaagatg acttagaaga
      961 agatgtttct gatgacgtct gagtggttac atgaggttcg tgtgtcgtag atggacttgt
     1021 agtgtcgctg ggtggctggt cggacgatgt ggttgaagaa tgtgtgaatg cgcttgttaa
     1081 cgttgtacta gaagaagttg tagatacagt ggaagaagat gttgtagtga cggcttgtgt
     1141 agatgcgcta gtagaatatg tcgtactgct actagatgag actggtacac tggtagcagt
     1201 actagttaat gacatttcgg atgttgagga tgaactagag gatgttgtga ccatttctgc
     1261 ggaattagca agaacagtgg cgtggctgga ggttgattcc acggactggg aagagccaga
     1321 tgttgtcgaa ctttctgatt tagtggacga agaaccggat gttgtagatg atgaacttga
     1381 tgaagccgac aaacctgtct gcgagtgact aactggagtc gaaattatcg atgtactaga
     1441 tgtatactct gtcattgaag taggtgtttt agatgtttct gtacttgtcg aagtagaggt
     1501 agatttacta ggcgatgagg aaactgtcac atctgtaact ggagctgaac ttgtagtgga
     1561 agttgagatt gactttgttg tggtggttga agtcattgag gaagatgatg aagaagttga
     1621 tggtgtagtt gatacggtcg gtgatgatga aatttcctca gattgggaag atggggaaga
     1681 cgatgaactg gacgaagatg ttgtagttgg gtcagatgat tgtgaagcag aggaagaggt
     1741 actagaggtt gatactgatg cttctgtaga gggtgaggat gtttgagatt gaaaatacaa
     1801 attttcctta gcgtaatctg gaacatcgta tgggtatgat ccgccaccac cagaaccccc
     1861 tccaccggat tgagaccgac tacggttatc cacagaatca ggggataacg caggaaagaa
     1921 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1981 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2041 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2101 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2161 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2221 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2281 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2341 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2401 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2461 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2521 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2581 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2641 catgattgat cgggcacgta agaggttcca actttcacca taatgaaata agatcactac
     2701 cgggcgtatt ttttgagtta tcgagatttt caggagctaa ggaagctaaa atggagaaaa
     2761 aaatcactgg atataccacc gttgatatat cccaatggca tcgtaaagaa cattttgagg
     2821 catttcagtc agttgctcaa tgtacctata accagaccgt tcagctggat attacggcct
     2881 ttttaaagac cgtaaagaaa aataagcaca agttttatcc ggcctttatt cacattcttg
     2941 cccgcctgat gaatgctcat ccggaatttc gtatggcaat gaaagacggt gagctggtga
     3001 tatgggatag tgttcaccct tgttacaccg ttttccatga gcaaactgaa acgttttcat
     3061 cgctctggag tgaataccac gacgatttcc ggcagtttct acacatatat tcgcaagatg
     3121 tggcgtgtta cggtgaaaac ctggcctatt tccctaaagg gtttattgag aatatgtttt
     3181 tcgtttcagc caatccctgg gtgagtttca ccagttttga tttaaacgtg gccaatatgg
     3241 acaacttctt cgcccccgtt ttcaccatgg gcaaatatta tacgcaaggc gacaaggtgc
     3301 tgatgccgct ggcgattcag gttcatcatg ccgtttgtga tggcttccat gtcggcagaa
     3361 tgcttaatga attacaacag tactgcgatg agtggcaggg cggggcgtaa tttgatatcg
     3421 agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     3481 ttcagaacgc tcggttgccg ccgggcgttt tttattggtc tggtctcagc cactcgagtt
     3541 agatgatagc caaagacaac aacaagaaca aagcaccagc accagcacca gcgttaacag
     3601 agttagcaga accagcgtag gtagagattt caaccataga agattggatt tgggcgccct
     3661 ttgtactagg tgatgatgca acacttatgg atgagtgtgc tgtctgtgaa ctatacacct
     3721 tagaatttga cgaagattct ttagcggaat gggaggtgcc tgatgttact gatgtagtag
     3781 actccgtgga agtggatgtt gtagtgcttt cgtcaactgc
//
