LOCUS       sequence-231223-        3893 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 3893)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 3893)
  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: CBL5-uPIC
FEATURES             Location/Qualifiers
     source          1..3893
                     /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             1592..1612
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1619..1636
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1716..1962
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1977..2390
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        2397..2444
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2463..2837
                     /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      2903..3150
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(2919..2937)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3225..3813)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3305..3324)
                     /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 tatgggatgt gtttgcagca agcaattaga aggaagaagg caagaagaca tttcgcttct
     1021 tgcttctcaa acctttttta gcgaggctga ggttgaagtt ttgcatggat tgttcatcaa
     1081 actcacttct tgtcttagca acgacaatct tttgaccaaa gaaaaatttc aatttatctt
     1141 gatcaagaat accaaaaagc gcagtctttc tgcagaacgg atttttggat tattcgatat
     1201 gagaaacgat ggggctatag attttggaga gtttgtacac acactcaaca tttttcaccc
     1261 aaactcatca ccaagagata aggccatatt tgcatttaga ttatacgata ctcgtgagac
     1321 tggattcatt gaaccggagg aggtgaaaga gatgataata gacgttctag aggaatcaga
     1381 acttatgcta tccgagagta tcattgattc tatcgtgtct aagacatttg aagaggccga
     1441 ttggaaaaaa gatgggataa tagatttgga agaatgggag aattttgttg ccacctatcc
     1501 tttgacacta aagaacatga ctatcccttt cttgaaggac attccgagga ttttcccaac
     1561 ctttctccgg ggattaaacc tcagcagcgg cgagaacctg tacttccaat ctgtcgacca
     1621 tcatcatcat catcattgag tttgtagcct tagacatgac tgttcctcag ttcaagttgg
     1681 gcacttacga gaagaccggt cttgctagat tctaatcaag aggatgtcag aatgccattt
     1741 gcctgagaga tgcaggcttc atttttgata cttttttatt tgtaacctat atagtatagg
     1801 attttttttg tcattttgtt tcttctcgta cgagcttgct cctgatcagc ctatctcgca
     1861 gctgatgaat atcttgtggt aggggtttgg gaaaatcatt cgagtttgat gtttttcttg
     1921 gtatttccca ctcctcttca gagtacagaa gattaagtga gaccttcgtt tgtgcggatc
     1981 ccccacacac catagcttca aaatgtttct actccttttt tactcttcca gattttctcg
     2041 gactccgcgc atcgccgtac cacttcaaaa cacccaagca cagcatacta aatttcccct
     2101 ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa gaaaaaagag
     2161 accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt
     2221 ttcttgaaaa tttttttttt tgattttttt ctctttcgat gacctcccat tgatatttaa
     2281 gttaataaac ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt ctattacaac
     2341 tttttttact tcttgctcat tagaaagaaa gcatagcaat ctaatctaag ggcggtgttg
     2401 acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg aggaactaaa
     2461 ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg
     2521 tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac gacttcgccg
     2581 gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag gtggtgccgg
     2641 acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg
     2701 aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag atcggcgagc
     2761 agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg cacttcgtgg
     2821 ccgaggagca ggactgacac gtccgacgcg gcccgacggg tccgaggcct cggagatccg
     2881 tccccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca ttcacgccct
     2941 ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt ctaggtccct
     3001 atttattttt ttatagttat gttagtatta agaacgttat ttatatttca aatttttctt
     3061 ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt gcttgagaag
     3121 gttttgggac gctcgaaggc tttaatttgc aagctggaga ccaacatgtg agcaaaaggc
     3181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3241 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3301 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3361 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3421 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3481 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3541 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3601 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3661 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3721 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3781 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3841 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag atc
//
