LOCUS       sequence-173094-        3983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducible expression of CeTRIC-B2 in P.Pichia strains.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3983)
  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 3983)
  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: pPICZC_CeTRIC-B2_Del
FEATURES             Location/Qualifiers
     source          1..3983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(215..262)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(269..682)
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     terminator      complement(697..943)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     CDS             complement(1023..1040)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        complement(1809..2747)
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
     rep_origin      2829..3417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3318..3337
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3492..3739)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     3705..3723
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     CDS             complement(join(3805..3983,1..196))
                     /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 cccaggccag ggtgttgtcc ggcaccacct ggtcctggac cgcgctgatg aacagggtca
       61 cgtcgtcccg gaccacaccg gcgaagtcgt cctccacgaa gtcccgggag aacccgagcc
      121 ggtcggtcca gaactcgacc gctccggcga cgtcgcgcgc ggtgagcacc ggaacggcac
      181 tggtcaactt ggccatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact
      241 atgccgatga ttaattgtca acaccgccct tagattagat tgctatgctt tctttctaat
      301 gagcaagaag taaaaaaagt tgtaatagaa caagaaaaat gaaactgaaa cttgagaaat
      361 tgaagaccgt ttattaactt aaatatcaat gggaggtcat cgaaagagaa aaaaatcaaa
      421 aaaaaaaatt ttcaagaaaa agaaacgtga taaaaatttt tattgccttt ttcgacgaag
      481 aaaaagaaac gaggcggtct cttttttctt ttccaaacct ttagtacggg taattaacga
      541 caccctagag gaagaaagag gggaaattta gtatgctgtg cttgggtgtt ttgaagtggt
      601 acggcgatgc gcggagtccg agaaaatctg gaagagtaaa aaaggagtag aaacattttg
      661 aagctatggt gtgtggggga tccgcacaaa cgaaggtctc acttaatctt ctgtactctg
      721 aagaggagtg ggaaatacca agaaaaacat caaactcgaa tgattttccc aaacccctac
      781 cacaagatat tcatcagctg cgagataggc tgatcaggag caagctcgta cgagaagaaa
      841 caaaatgaca aaaaaaatcc tatactatat aggttacaaa taaaaaagta tcaaaaatga
      901 agcctgcatc tctcaggcaa atggcattct gacatcctct tgattagaat ctagcaagac
      961 cggtcttctc gtaagtgccc aacttgaact gaggaacagt catgtctaag gctacaaact
     1021 caatgatgat gatgatgatg aagaacgttt tcaataggca tcaagacgtc atgaacacta
     1081 aggcacaatg atgccaactt gaagtagatg aaaaatccga caacacaaag ataaaccaaa
     1141 tcgtgtgggg cggtaacgta catggaatgt ctctccaatg tgaagacgat agaggcaata
     1201 acacaagcct ttgtagtaaa ggaaggtctc aagatttcgt ggttagttgg atgccaggta
     1261 cctctagcca actgctcaac aattttgaca acaccacttc ctgcaccctt ggcgactcca
     1321 acaaggattt gaaccaaata tgattctggg tagattctga cagcgtgttt aacaccagct
     1381 gcaatcttat gtgttctctg gacctctttc aaaacagaaa gtcccaactt aacagggaac
     1441 caagtagcaa gtctgaagac aacgtcaaat ggactataga agaccaagta ccaaacgatt
     1501 gaaccaagca aaatatcagc gtgttgtttc aaaggtgaga taattggctc tccaagcaaa
     1561 aagcaactca agaatgatcc agcaaaggac ataagcatgc aggacaacca acaagagaaa
     1621 ggatgctttc tggaaaaaga aatggcgtct ggtcccaaat cgtgtctgac agaaagacaa
     1681 gtcaaaacat aatgggcagc atcaaagtat ggatacaact tcaacttctg gacgtaacca
     1741 cctgcgtcaa tcaatgtgtc gtgatcaata gaccactcat ctggaaccca acccatggtg
     1801 gaattcctcg tttcgaataa ttagttgttt tttgatcttc tcaagttgtc gttaaaagtc
     1861 gttaaaatca aaagcttgtc aattggaacc agtcgcaatt atgaaagtaa gctaataatg
     1921 atgataaaaa aaaaggttta agacagggca gcttccttct gtttatatat tgctgtcaag
     1981 taggggttag aacagttaaa ttttgatcat gaacgttagg ctatcagcag tattcccacc
     2041 agaatcttgg aagcatacaa tgtggagaca atgcataatc atccaaaaag cgggtgtttc
     2101 cccatttgcg tttcggcaca ggtgcaccgg ggttcagaag cgatagagag actgcgctaa
     2161 gcattaatga gattattttt gagcattcgt caatcaatac caaacaagac aaacggtatg
     2221 ccgacttttg gaagtttctt tttgaccaac tggccgttag catttcaacg aaccaaactt
     2281 agttcatctt ggatgagatc acgcttttgt catattaggt tccaagacag cgtttaaact
     2341 gtcagttttg ggccatttgg ggaacatgaa actatttgac cccacactca gaaagccctc
     2401 atctggagtg atgttcgggt gtaatgcgga gcttgttgca ttcggaaata aacaaacatg
     2461 aacctcgcca ggggggccag gatagacagg ctaataaagt catggtgtta gtagcctaat
     2521 agaaggaatt ggaatgagcg agctccaatc aagcccaata actgggctgg tttttcgatg
     2581 gcaaaagtgg gtgttgagga gaagaggagt ggaggtcctg cgtttgcaac ggtctgctgc
     2641 tagtgtatcc cctcctgttg cgtttggcac ttatgtgtga gaatggacct gtggatgtcg
     2701 gatggcaaaa aggtttcatt caacctttcg tctttggatg ttagatctga tctcatgacc
     2761 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2821 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2881 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2941 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3001 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3061 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3121 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3181 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3241 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3301 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3361 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3421 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttg
     3481 gtctccagct tgcaaattaa agccttcgag cgtcccaaaa ccttctcaag caaggttttc
     3541 agtataatgt tacatgcgta cacgcgtctg tacagaaaaa aaagaaaaat ttgaaatata
     3601 aataacgttc ttaatactaa cataactata aaaaaataaa tagggaccta gacttcaggt
     3661 tgtctaactc cttccttttc ggttagagcg gatgtggggg gagggcgtga atgtaagcgt
     3721 gacataacta attacatgat atcgacaaag gaaaaggggg acggatctcc gaggcctcgg
     3781 acccgtcggg ccgcgtcgga cgtgtcagtc ctgctcctcg gccacgaagt gcacgcagtt
     3841 gccggccggg tcgcgcaggg cgaactcccg cccccacggc tgctcgccga tctcggtcat
     3901 ggccggcccg gaggcgtccc ggaagttcgt ggacacgacc tccgaccact cggcgtacag
     3961 ctcgtccagg ccgcgcaccc aca
//
