LOCUS       sequence-231212-        3683 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  protein expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3683)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 3683)
  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: SbMBP
FEATURES             Location/Qualifiers
     source          1..3683
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..940
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     CDS             1382..1402
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1409..1426
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1506..1752
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1767..2180
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2187..2234
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2253..2627
                     /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      2693..2940
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2709..2727)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3015..3603)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3095..3114)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag
       61 gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt
      121 tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc
      181 agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta
      241 acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta
      301 tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg
      361 agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct
      421 gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg
      481 ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt
      541 cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct
      601 ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct
      661 ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact
      721 gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat
      781 atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt
      841 actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga
      901 caacttgaga agatcaaaaa acaactaatt attcgaaacg aggaattcgc tgaggcttaa
      961 tatgtccaag gtcttcaccc tcgaggacgt cgccaagcac aacaccaagg aggattgctg
     1021 gctcatcatc ggcggcaagg tatatgatgt gacgaagttt ttggaagacc acccaggggg
     1081 tgatgatgtt ctgttgtctt caactggtaa ggatgcaact gatgatttcg aggatgtcgg
     1141 gcacagcaac actgcccgtg caatgatgga tgagtacttg gtcggtgaaa ttgacgcatc
     1201 gacgatccct tctaggacaa agtatgttcc ccccaagcaa ccgcactaca accaggacaa
     1261 gacccccgag tttgttatca agatcctcca gtttttggtt cccctggcaa tcctgggcct
     1321 ggctgttgcc gttaggatgt acaccaagtc ggagtctgct gcattaaacc tcagcagcgg
     1381 cgagaacctg tacttccaat ctgtcgacca tcatcatcat catcattgag tttgtagcct
     1441 tagacatgac tgttcctcag ttcaagttgg gcacttacga gaagaccggt cttgctagat
     1501 tctaatcaag aggatgtcag aatgccattt gcctgagaga tgcaggcttc atttttgata
     1561 cttttttatt tgtaacctat atagtatagg attttttttg tcattttgtt tcttctcgta
     1621 cgagcttgct cctgatcagc ctatctcgca gctgatgaat atcttgtggt aggggtttgg
     1681 gaaaatcatt cgagtttgat gtttttcttg gtatttccca ctcctcttca gagtacagaa
     1741 gattaagtga gaccttcgtt tgtgcggatc ccccacacac catagcttca aaatgtttct
     1801 actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac cacttcaaaa
     1861 cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc gttaattacc
     1921 cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa
     1981 aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt tgattttttt
     2041 ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt
     2101 tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa
     2161 gcatagcaat ctaatctaag ggcggtgttg acaattaatc atcggcatag tatatcggca
     2221 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     2281 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     2341 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     2401 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     2461 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     2521 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     2581 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtccgacgcg
     2641 gcccgacggg tccgaggcct cggagatccg tccccctttt cctttgtcga tatcatgtaa
     2701 ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa
     2761 ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta
     2821 agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt
     2881 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc
     2941 aagctggaga ccaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     3001 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     3061 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     3121 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     3181 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     3241 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     3301 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     3361 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     3421 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     3481 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     3541 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3601 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3661 taagggattt tggtcatgag atc
//
