LOCUS       sequence-225005-        4945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4945)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4945)
  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"
COMMENT     Sequence Label: ZmMATE2
FEATURES             Location/Qualifiers
     source          1..4945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1674..1697
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1698..1721
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             1731..1748
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1828..2074
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2089..2502
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2509..2556
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2575..2949
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3015..3262
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3031..3049)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3337..3925)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3417..3436)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4007..4945
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
ORIGIN
        1 aggaattcat ggaggataac ggtgctgctg ccgcagctgc tgctccagct acgtctatgc
       61 ctgccgagaa gagagtggca gtcattcaag gcgatgcccc cccagagcct gcagccggat
      121 tattgccatg cggtccacga aaaacaggct tgcacttatt cgtaatgaac attaggtccg
      181 tctttaagct ggacggtctg ggtagtgagg tattacgtat cgcagttcct gcatccttgg
      241 ccttggcagc cgatcctttg gcttctctag tggacacggc cttcatcggt agacttggtt
      301 ctgtcgaaat tgctgctgtc ggggttagta ttgccatctt taaccaagtg tccaaagtct
      361 gtatatatcc acttgtctca gtgactacta gttttgtagc agaagaagac gcaattattt
      421 caaaagccgt gagaggtaat tcttcacagg aggaagatgt cgagaaggca tctcatgtgg
      481 gatttgaccc tgaaacatca aacctgcacg cttctggtcc cgctggcatg gcagaatgtg
      541 ttaactcctg tatcccaact gaatgtgccg cagatccttc cggtcgtcaa ggtaggtgcg
      601 aaaaacgtta cgtgccctct gtgacttccg ctttgattgt aggttccatt ttgggtttat
      661 tgcaagctgt gttcttggtt ctttctgcaa gatttgtact aaatatcatg ggagtcaaat
      721 ctggatctcc aatgcagggt cccgctgtga gatatttaac cattagatcc ttaggtgccc
      781 ctgccgtgtt gttgtccctt gcaatgcaag gtgtgtttcg tggttttaag gacactaaaa
      841 ctcccctata tgccactgtt gtaggcgacg ctgctaatat cattcttgac cctatcttga
      901 tgtttgtgtg tcacatgggg gtcacaggtg ctgcaattgc ccatgttgtt tcccagtaca
      961 tgatcacatt gatcttgttg tgtagactgg tccaacgagt ccatgtcatc ccaccaagta
     1021 ttaagtcttt aaaattcggc cgatttcttg gttgtggttt tttgttgttg gctcgtgttg
     1081 ttgccgttac cttttgcgta acacttgctg catccctggc agctagacat ggtccaaccg
     1141 taatggctgg attccaaatt tgttgccaat tatggctggc tacttctttg ctagctgatg
     1201 gacttgctgt tgctggtcaa gccgtgctgg cctccgcttt cgccaaaaat gattcaaaaa
     1261 aagttgctgc tgctacatca agagtattgc aattgagtat agttctaggt atgggcttaa
     1321 ctgtcgtttt gggtttggcc atgcgtttcg gcgccggaat tttcacctct gacgtgcctg
     1381 tgattcaggt cattcacaga ggaataccct ttgttgctgg cacacagacc atcaactctt
     1441 tggccttcgt ctttgacggt attaacttcg gagctagtga ttacagatat tccgcttact
     1501 ctatggttgc tgtcgcttca gtttctatcc catgtctgtt atatttatct gctcataacg
     1561 gctttattgg aatttggata gctttgacaa tctatatgtc acttagaact atcgcttcta
     1621 cttggagaat gggggccgct cgtggtccat ggacatttct gaggaacggt accttggagg
     1681 ttttgttcca gggtccacat catcaccacc atcaccatca ttgagtcgac catcatcatc
     1741 atcatcattg agtttgtagc cttagacatg actgttcctc agttcaagtt gggcacttac
     1801 gagaagaccg gtcttgctag attctaatca agaggatgtc agaatgccat ttgcctgaga
     1861 gatgcaggct tcatttttga tactttttta tttgtaacct atatagtata ggattttttt
     1921 tgtcattttg tttcttctcg tacgagcttg ctcctgatca gcctatctcg cagctgatga
     1981 atatcttgtg gtaggggttt gggaaaatca ttcgagtttg atgtttttct tggtatttcc
     2041 cactcctctt cagagtacag aagattaagt gagaccttcg tttgtgcgga tcccccacac
     2101 accatagctt caaaatgttt ctactccttt tttactcttc cagattttct cggactccgc
     2161 gcatcgccgt accacttcaa aacacccaag cacagcatac taaatttccc ctctttcttc
     2221 ctctagggtg tcgttaatta cccgtactaa aggtttggaa aagaaaaaag agaccgcctc
     2281 gtttcttttt cttcgtcgaa aaaggcaata aaaattttta tcacgtttct ttttcttgaa
     2341 aatttttttt tttgattttt ttctctttcg atgacctccc attgatattt aagttaataa
     2401 acggtcttca atttctcaag tttcagtttc atttttcttg ttctattaca acttttttta
     2461 cttcttgctc attagaaaga aagcatagca atctaatcta agggcggtgt tgacaattaa
     2521 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
     2581 aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc ggtcgagttc
     2641 tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc cggtgtggtc
     2701 cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc ggacaacacc
     2761 ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc ggaggtcgtg
     2821 tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga gcagccgtgg
     2881 gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt ggccgaggag
     2941 caggactgac acgtccgacg cggcccgacg ggtccgaggc ctcggagatc cgtccccctt
     3001 ttcctttgtc gatatcatgt aattagttat gtcacgctta cattcacgcc ctccccccac
     3061 atccgctcta accgaaaagg aaggagttag acaacctgaa gtctaggtcc ctatttattt
     3121 ttttatagtt atgttagtat taagaacgtt atttatattt caaatttttc ttttttttct
     3181 gtacagacgc gtgtacgcat gtaacattat actgaaaacc ttgcttgaga aggttttggg
     3241 acgctcgaag gctttaattt gcaagctgga gaccaacatg tgagcaaaag gccagcaaaa
     3301 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3361 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3421 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3481 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     3541 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     3601 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     3661 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     3721 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     3781 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     3841 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     3901 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     3961 tcagtggaac gaaaactcac gttaagggat tttggtcatg agatcagatc taacatccaa
     4021 agacgaaagg ttgaatgaaa cctttttgcc atccgacatc cacaggtcca ttctcacaca
     4081 taagtgccaa acgcaacagg aggggataca ctagcagcag accgttgcaa acgcaggacc
     4141 tccactcctc ttctcctcaa cacccacttt tgccatcgaa aaaccagccc agttattggg
     4201 cttgattgga gctcgctcat tccaattcct tctattaggc tactaacacc atgactttat
     4261 tagcctgtct atcctggccc ccctggcgag gttcatgttt gtttatttcc gaatgcaaca
     4321 agctccgcat tacacccgaa catcactcca gatgagggct ttctgagtgt ggggtcaaat
     4381 agtttcatgt tccccaaatg gcccaaaact gacagtttaa acgctgtctt ggaacctaat
     4441 atgacaaaag cgtgatctca tccaagatga actaagtttg gttcgttgaa atgctaacgg
     4501 ccagttggtc aaaaagaaac ttccaaaagt cggcataccg tttgtcttgt ttggtattga
     4561 ttgacgaatg ctcaaaaata atctcattaa tgcttagcgc agtctctcta tcgcttctga
     4621 accccggtgc acctgtgccg aaacgcaaat ggggaaacac ccgctttttg gatgattatg
     4681 cattgtctcc acattgtatg cttccaagat tctggtggga atactgctga tagcctaacg
     4741 ttcatgatca aaatttaact gttctaaccc ctacttgaca gcaatatata aacagaagga
     4801 agctgccctg tcttaaacct ttttttttat catcattatt agcttacttt cataattgcg
     4861 actggttcca attgacaagc ttttgatttt aacgactttt aacgacaact tgagaagatc
     4921 aaaaaacaac taattattcg aaacg
//
