LOCUS       sequence-231296-        4652 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 4652)
  TITLE     Chang lab yeast expression plasmids
REFERENCE   2  (bases 1 to 4652)
  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: CIPK2-uPIC
FEATURES             Location/Qualifiers
     source          1..4652
                     /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             2351..2371
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2378..2395
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2475..2721
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        2736..3149
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        3156..3203
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3222..3596
                     /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      3662..3909
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3678..3696)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(3984..4572)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4064..4083)
                     /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 tatggagaac aaaccaagtg tattaactga aagatatgaa gttgggaggt tattgggtca
     1021 aggtactttt gctaaagtgt attttggaag gagtaatcat accaacgaga gtgtagctat
     1081 caagatgatt gacaaggaca aagttatgag agtcgggctt agccagcaaa tcaagcgaga
     1141 gatctctgta atgaggatcg ctaaacaccc gaatgtcgtt gagttatacg aggttatggc
     1201 tacaaagtca aggatttact ttgttattga gtattgtaaa ggtggtgagc ttttcaacaa
     1261 ggttgcaaaa ggaaaactta aagaagatgt tgcttggaag tatttttatc agcttattag
     1321 tgcggttgat ttttgtcaca gccgcggagt ttatcaccgc gacattaagc cggaaaatct
     1381 tttgttggat gacaatgata atcttaaggt atctgatttt ggtttaagtg cgcttgctga
     1441 ttgcaagcgg caagatggtc ttctacatac aacttgtggt acacctgctt atgttgcgcc
     1501 cgaggttatt aaccgaaaag gatacgaggg tacgaaagcg gatatttggt cttgtggtgt
     1561 tgttttgttt gttcttttgg ctggttatct tcctttccat gacactaatc ttatggagat
     1621 gtataggaag ataggtaaag cagacttcaa gtgtcccagc tggtttgctc ctgaggtaaa
     1681 gagactattg tgtaagatgt tggaccctaa ccatgagact agaatcacta ttgcaaaaat
     1741 caaggagagt tcttggttca gaaagggttt gcatttgaag caaaaaaaga tggaaaagat
     1801 ggagaaacaa caagtcagag aggctactaa tcccatggaa gctggaggtt caggccaaaa
     1861 tgaaaacgga gaaaaccacg agccgcctcg acttgctacc ttgaatgctt tcgatatcat
     1921 cgccttgtct acggggtttg gtctggcagg actttttggg gatgtatatg acaagcgaga
     1981 atctaggttc gcgtcgcaaa aacctgcttc agagatcatt tctaagctag tagaggttgc
     2041 caagtgcctg aagctgaaga taagaaagca aggcgcaggc ttgttcaaac tggaaagagt
     2101 aaaggaagga aaaaacggaa ttttgacgat ggatgcagag atattccaag tgacgccgac
     2161 gtttcacctg gtggaagtga agaaatgtaa tggagataca atggagtatc agaagttagt
     2221 ggaggaggat cttaggcctg ctttggcaga tattgtttgg gtttggcaag gcgagaagga
     2281 gaaagaggag cagttactgc aggatgaaca aggagagcaa gaaccatcag cattaaacct
     2341 cagcagcggc gagaacctgt acttccaatc tgtcgaccat catcatcatc atcattgagt
     2401 ttgtagcctt agacatgact gttcctcagt tcaagttggg cacttacgag aagaccggtc
     2461 ttgctagatt ctaatcaaga ggatgtcaga atgccatttg cctgagagat gcaggcttca
     2521 tttttgatac ttttttattt gtaacctata tagtatagga ttttttttgt cattttgttt
     2581 cttctcgtac gagcttgctc ctgatcagcc tatctcgcag ctgatgaata tcttgtggta
     2641 ggggtttggg aaaatcattc gagtttgatg tttttcttgg tatttcccac tcctcttcag
     2701 agtacagaag attaagtgag accttcgttt gtgcggatcc cccacacacc atagcttcaa
     2761 aatgtttcta ctcctttttt actcttccag attttctcgg actccgcgca tcgccgtacc
     2821 acttcaaaac acccaagcac agcatactaa atttcccctc tttcttcctc tagggtgtcg
     2881 ttaattaccc gtactaaagg tttggaaaag aaaaaagaga ccgcctcgtt tctttttctt
     2941 cgtcgaaaaa ggcaataaaa atttttatca cgtttctttt tcttgaaaat tttttttttt
     3001 gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg gtcttcaatt
     3061 tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt cttgctcatt
     3121 agaaagaaag catagcaatc taatctaagg gcggtgttga caattaatca tcggcatagt
     3181 atatcggcat agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg
     3241 ccgttccggt gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc
     3301 tcgggttctc ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga
     3361 ccctgttcat cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt
     3421 gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc
     3481 gggacgcctc cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg
     3541 ccctgcgcga cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg
     3601 tccgacgcgg cccgacgggt ccgaggcctc ggagatccgt cccccttttc ctttgtcgat
     3661 atcatgtaat tagttatgtc acgcttacat tcacgccctc cccccacatc cgctctaacc
     3721 gaaaaggaag gagttagaca acctgaagtc taggtcccta tttatttttt tatagttatg
     3781 ttagtattaa gaacgttatt tatatttcaa atttttcttt tttttctgta cagacgcgtg
     3841 tacgcatgta acattatact gaaaaccttg cttgagaagg ttttgggacg ctcgaaggct
     3901 ttaatttgca agctggagac caacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3961 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4021 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4081 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4141 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4201 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4261 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4321 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4381 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4441 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4501 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4561 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4621 aactcacgtt aagggatttt ggtcatgaga tc
//
