LOCUS       sequence-231503-        4763 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 4763)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4763)
  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: AtALMT-uPIC
FEATURES             Location/Qualifiers
     source          1..4763
                     /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             complement(1067..1078)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             2462..2482
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2489..2506
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2586..2832
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2847..3260
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3267..3314
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3333..3707
                     /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      3773..4020
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3789..3807)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(4095..4683)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4175..4194)
                     /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 tatggagaaa gtgagagaga tagtgagaga agggattaga gtagggaatg aagacccgag
     1021 aagaattatt catgctttca aagtgggact tgctcttgtt ttggtctctt ccttctatta
     1081 ttaccaacct ttcggtcctt tcaccgatta cttcggtatc aatgcaatgt gggctgtcat
     1141 gaccgtcgtt gttgtctttg aattctccgt aggggctaca cttgggaaag gactaaatag
     1201 aggagtggca acattagtag caggaggact aggaattgga gctcatcaac tagcaagatt
     1261 gtcgggtgca acagtggaac ctattctttt ggtcatgttg gtctttgtgc aagctgcgtt
     1321 gtcgacgttc gtaaggtttt tcccatgggt aaagacaaag ttcgattatg ggatattgat
     1381 attcatattg acgttcgcgc tgatatcgtt gtcggggttt agagacgaag agataatgga
     1441 tttggcagaa agcagactat cgacagtggt gattggagga gtcagttgca tccttatctc
     1501 tatctttgtc tgtcctgttt gggctggaca agatctacat tctctccttg cctctaactt
     1561 tgacactctc tcccacttcc ttcaagattt tggggacgaa tattttgaag cgagagagaa
     1621 gggggactac aaagtagtgg agaagaggaa aaagaatctt gagagatata aaagtgttct
     1681 tgactccaaa agcgatgaag aagctttggc taattacgca gaatgggaac cgcctcatgg
     1741 acaatttagg tttaggcatc catggaaaca atatgttgcc gttggtgcat tactccggca
     1801 gtgtgcctac aggattgatg ccttgaattc atacatcaac tcagattttc agatcccagt
     1861 ggacataaaa aagaaattag aaacaccatt aagaagaatg agctcggaat cgggaaactc
     1921 aatgaaagaa atgtccattt cactaaaaca aatgataaaa tcatcttctt ctgatatcca
     1981 tgtctcaaac tcacaagctg catgcaaatc tctttctact ttacttaaat caggcatctt
     2041 gaacgatgtt gagcctcttc aaatgatctc attgatgacg actgtctcca tgctcataga
     2101 tattgttaac ttaaccgaaa agatctcgga atctgtacac gagcttgcat ccgctgcaag
     2161 atttaagaac aagatgaggc cgaccgtgct atacgagaag tcggattccg gaagcattgg
     2221 tcgtgccatg ccaatagatt ctcatgaaga tcatcatgtt gtcacagttt tacatgacgt
     2281 tgataatgat agatcaaaca atgttgatga ttcacgtgga gggagttcac aggactcatg
     2341 tcatcatgtt gccataaaga ttgttgatga taactcaaac catgaaaaac atgaagatgg
     2401 tgaaatacat gtacatacat taagtaatgg gcatcttcag gcattaaacc tcagcagcgg
     2461 cgagaacctg tacttccaat ctgtcgacca tcatcatcat catcattgag tttgtagcct
     2521 tagacatgac tgttcctcag ttcaagttgg gcacttacga gaagaccggt cttgctagat
     2581 tctaatcaag aggatgtcag aatgccattt gcctgagaga tgcaggcttc atttttgata
     2641 cttttttatt tgtaacctat atagtatagg attttttttg tcattttgtt tcttctcgta
     2701 cgagcttgct cctgatcagc ctatctcgca gctgatgaat atcttgtggt aggggtttgg
     2761 gaaaatcatt cgagtttgat gtttttcttg gtatttccca ctcctcttca gagtacagaa
     2821 gattaagtga gaccttcgtt tgtgcggatc ccccacacac catagcttca aaatgtttct
     2881 actccttttt tactcttcca gattttctcg gactccgcgc atcgccgtac cacttcaaaa
     2941 cacccaagca cagcatacta aatttcccct ctttcttcct ctagggtgtc gttaattacc
     3001 cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa
     3061 aggcaataaa aatttttatc acgtttcttt ttcttgaaaa tttttttttt tgattttttt
     3121 ctctttcgat gacctcccat tgatatttaa gttaataaac ggtcttcaat ttctcaagtt
     3181 tcagtttcat ttttcttgtt ctattacaac tttttttact tcttgctcat tagaaagaaa
     3241 gcatagcaat ctaatctaag ggcggtgttg acaattaatc atcggcatag tatatcggca
     3301 tagtataata cgacaaggtg aggaactaaa ccatggccaa gttgaccagt gccgttccgg
     3361 tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg ctcgggttct
     3421 cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg accctgttca
     3481 tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg
     3541 gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct
     3601 ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc gccctgcgcg
     3661 acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgacac gtccgacgcg
     3721 gcccgacggg tccgaggcct cggagatccg tccccctttt cctttgtcga tatcatgtaa
     3781 ttagttatgt cacgcttaca ttcacgccct ccccccacat ccgctctaac cgaaaaggaa
     3841 ggagttagac aacctgaagt ctaggtccct atttattttt ttatagttat gttagtatta
     3901 agaacgttat ttatatttca aatttttctt ttttttctgt acagacgcgt gtacgcatgt
     3961 aacattatac tgaaaacctt gcttgagaag gttttgggac gctcgaaggc tttaatttgc
     4021 aagctggaga ccaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4081 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4141 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4201 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4261 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4321 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4381 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4441 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4501 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4561 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4621 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4681 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4741 taagggattt tggtcatgag atc
//
