LOCUS       sequence-173093-        3978 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of CeTRIC-B1 in P.Pichia strains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3978)
  AUTHORS   Yang H, Hu M, Guo J, Ou X, Cai T, Liu Z
  TITLE     Pore architecture of TRIC channels and insights into their gating 
            mechanism.
  JOURNAL   Nature. 2016 Oct 27;538(7626):537-541. doi: 10.1038/nature19767. 
            Epub 2016 Oct 3.
  PUBMED    27698420
REFERENCE   2  (bases 1 to 3978)
  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"; doi: 
            "10.1038/nature19767"; journalName: "Nature"; date: "2016-10-27- 
            27"; volume: "538"; issue: "7626"; pages: "537-541"
COMMENT     Sequence Label: pPICZA_CeTRIC-B1_Del
FEATURES             Location/Qualifiers
     source          1..3978
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(187..234)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(241..654)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(669..915)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(994..1011)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(1776..2714)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      2796..3384
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3285..3304
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3459..3706)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3672..3690
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(join(3772..3978,1..168))
                     /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"
ORIGIN
        1 ctggtcctgg accgcgctga tgaacagggt cacgtcgtcc cggaccacac cggcgaagtc
       61 gtcctccacg aagtcccggg agaacccgag ccggtcggtc cagaactcga ccgctccggc
      121 gacgtcgcgc gcggtgagca ccggaacggc actggtcaac ttggccatgg tttagttcct
      181 caccttgtcg tattatacta tgccgatata ctatgccgat gattaattgt caacaccgcc
      241 cttagattag attgctatgc tttctttcta atgagcaaga agtaaaaaaa gttgtaatag
      301 aacaagaaaa atgaaactga aacttgagaa attgaagacc gtttattaac ttaaatatca
      361 atgggaggtc atcgaaagag aaaaaaatca aaaaaaaaaa ttttcaagaa aaagaaacgt
      421 gataaaaatt tttattgcct ttttcgacga agaaaaagaa acgaggcggt ctcttttttc
      481 ttttccaaac ctttagtacg ggtaattaac gacaccctag aggaagaaag aggggaaatt
      541 tagtatgctg tgcttgggtg ttttgaagtg gtacggcgat gcgcggagtc cgagaaaatc
      601 tggaagagta aaaaaggagt agaaacattt tgaagctatg gtgtgtgggg gatccgcaca
      661 aacgaaggtc tcacttaatc ttctgtactc tgaagaggag tgggaaatac caagaaaaac
      721 atcaaactcg aatgattttc ccaaacccct accacaagat attcatcagc tgcgagatag
      781 gctgatcagg agcaagctcg tacgagaaga aacaaaatga caaaaaaaat cctatactat
      841 ataggttaca aataaaaaag tatcaaaaat gaagcctgca tctctcaggc aaatggcatt
      901 ctgacatcct cttgattaga atctagcaag accggtcttc tcgtaagtgc ccaacttgaa
      961 ctgaggaaca gtcatgtcta aggctaaaac tcaatgatga tgatgatgat ggaacaaatt
     1021 ctcaatagga gcaaatgggt cagtgacgtg caagataaca gcagaaagct tgaagtagac
     1081 aaagaaacca acaatgacaa gataaaccaa atcatgaggg gctgtcaagt aagttcctga
     1141 tttttcaaga gccaaaacgg atgcggcgac aacacaggcc ttggtagcaa aggaaggtct
     1201 aagcaactca ttgtgtgttg gcaaccagac tcctctaaca agctgttcca aagttctaac
     1261 aataccagat cctgcacctt tggcagtacc aaccaagact tgaacgatat aactgtttgg
     1321 gtaaagttta gctgcgtgtg agactccatg ggaaaccttg tatgctcttt taacttcctt
     1381 catgacggcc aaaacacact tgacaggtgt aattttagcg atcttgtaga ttccgtcaaa
     1441 tggtgcatag aaaaccaagt accagataat agtagcaagg ataatgtcat catgtctttt
     1501 gaaaggtgca ataactggct ctccaagcaa aaagttggcc aagaaagcat ctgcaaagca
     1561 catcaacatg gaagaaagcc aacaagacaa tgggtgcttt ctagagaaaa tactggcagc
     1621 tgaacccaag tcatctctga cttcgataat cataagcaaa taatgagcaa catcgaaata
     1681 agggtacatt ttcaatctat gcaactgggc acctgcgtca agaagaatct cctggtcaag
     1741 ctggaaggac tctgggacaa ccatggtgaa ttcctcgttt cgaataatta gttgtttttt
     1801 gatcttctca agttgtcgtt aaaagtcgtt aaaatcaaaa gcttgtcaat tggaaccagt
     1861 cgcaattatg aaagtaagct aataatgatg ataaaaaaaa aggtttaaga cagggcagct
     1921 tccttctgtt tatatattgc tgtcaagtag gggttagaac agttaaattt tgatcatgaa
     1981 cgttaggcta tcagcagtat tcccaccaga atcttggaag catacaatgt ggagacaatg
     2041 cataatcatc caaaaagcgg gtgtttcccc atttgcgttt cggcacaggt gcaccggggt
     2101 tcagaagcga tagagagact gcgctaagca ttaatgagat tatttttgag cattcgtcaa
     2161 tcaataccaa acaagacaaa cggtatgccg acttttggaa gtttcttttt gaccaactgg
     2221 ccgttagcat ttcaacgaac caaacttagt tcatcttgga tgagatcacg cttttgtcat
     2281 attaggttcc aagacagcgt ttaaactgtc agttttgggc catttgggga acatgaaact
     2341 atttgacccc acactcagaa agccctcatc tggagtgatg ttcgggtgta atgcggagct
     2401 tgttgcattc ggaaataaac aaacatgaac ctcgccaggg gggccaggat agacaggcta
     2461 ataaagtcat ggtgttagta gcctaataga aggaattgga atgagcgagc tccaatcaag
     2521 cccaataact gggctggttt ttcgatggca aaagtgggtg ttgaggagaa gaggagtgga
     2581 ggtcctgcgt ttgcaacggt ctgctgctag tgtatcccct cctgttgcgt ttggcactta
     2641 tgtgtgagaa tggacctgtg gatgtcggat ggcaaaaagg tttcattcaa cctttcgtct
     2701 ttggatgtta gatctgatct catgaccaaa atcccttaac gtgagttttc gttccactga
     2761 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     2821 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     2881 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     2941 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3001 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3061 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3121 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     3181 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     3241 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     3301 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     3361 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     3421 ttttgctggc cttttgctca catgttggtc tccagcttgc aaattaaagc cttcgagcgt
     3481 cccaaaacct tctcaagcaa ggttttcagt ataatgttac atgcgtacac gcgtctgtac
     3541 agaaaaaaaa gaaaaatttg aaatataaat aacgttctta atactaacat aactataaaa
     3601 aaataaatag ggacctagac ttcaggttgt ctaactcctt ccttttcggt tagagcggat
     3661 gtggggggag ggcgtgaatg taagcgtgac ataactaatt acatgatatc gacaaaggaa
     3721 aagggggacg gatctccgag gcctcggacc cgtcgggccg cgtcggacgt gtcagtcctg
     3781 ctcctcggcc acgaagtgca cgcagttgcc ggccgggtcg cgcagggcga actcccgccc
     3841 ccacggctgc tcgccgatct cggtcatggc cggcccggag gcgtcccgga agttcgtgga
     3901 cacgacctcc gaccactcgg cgtacagctc gtccaggccg cgcacccaca cccaggccag
     3961 ggtgttgtcc ggcaccac
//
