LOCUS       sequence-192045-        2795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  empty BB3 for insertion and overexpression of 1 gene in P. pastoris.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2795)
  AUTHORS   Prielhofer R, Barrero JJ, Steuer S, Gassler T, Zahrl R, Baumann K, 
            Sauer M, Mattanovich D, Gasser B, Marx H
  TITLE     GoldenPiCS: a Golden Gate-derived modular cloning system for applied
            synthetic biology in the yeast Pichia pastoris.
  JOURNAL   BMC Syst Biol. 2017 Dec 8;11(1):123. doi: 10.1186/s12918-017-0492-3.
  PUBMED    29221460
REFERENCE   2  (bases 1 to 2795)
  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"; journalName: "BMC Syst 
            Biol."; date: "2017-12-8"; volume: "11(1):123. doi"; pages: " 
            10.1186/s12918-017-0492-3"
COMMENT     Sequence Label: BB3aZ_14*
FEATURES             Location/Qualifiers
     source          1..2795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(62..650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(142..161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    798..831
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        848..1255
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1262..1309
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1328..1702
                     /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      1712..1909
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    1932..1965
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 cgctgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
       61 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
      121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
      181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
      241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
      301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
      361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
      421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
      481 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
      541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
      601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
      661 tgatcttttc tacggggtgg agcgcaaaaa accccgcccc tgacagggcg gggttttttc
      721 gcgatcggag gcgtcttcgg atccctcgag gaagacgccg ctccggcatg gtacgctgca
      781 ggtcgacaac ccttaatata acttcgtata atgtatgcta tacgaagtta ttaggtctag
      841 atcggtacca cacaccatag cttcaaaatg tttctactcc ttttttactc ttccagattt
      901 tctcggactc cgcgcatcgc cgtaccactt caaaacaccc aagcacagca tactaaattt
      961 cccctctttc ttcctctagg gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa
     1021 aagcgaccgc ctcgtttctt tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt
     1081 tctttttctt gaaaattttt ttttttgatt tttttctctt tcgatgacct cccattgata
     1141 tttaagttaa taaacggtca tcaatttctc aagtttcagt ttcatttttc ttgttctatt
     1201 acaacttttt ttacttcttg ctcattagaa agaaagcata gcaatctaat ctaagggcgg
     1261 tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa
     1321 ctaaaccatg gccaagttga ccagtgccgt tccggtgcta accgcgcgcg acgtcgccgg
     1381 agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg aggacgactt
     1441 cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg accaggtggt
     1501 gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg
     1561 gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga ccgagatcgg
     1621 cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact gcgtgcactt
     1681 cgtggccgag gagcaggact gatcagtact tactgacaat aaaaagattc ttgttttcaa
     1741 gaacttgtca tttgtatagt ttttttatat tgtagttgtt ctattttaat caaatgttag
     1801 cgtgatttat attttttttc gcctcgacat catctgccca gatgcgaagt taagtgcgca
     1861 gaaagtaata tcatgcgtca atcgtatgtg aatgctggtc gctatactgg taccattcga
     1921 gaacccttaa tataacttcg tataatgtat gctatacgaa gttattaggt gatatcagat
     1981 ccactgcttc agcaacgttg tcactgaagt tggcatcagt atctacaaac cctacatacc
     2041 tcttgaaggt ccagaaggta agcatttgcc cgctccattc ctacccgaca tatccactct
     2101 acattttata gttcaaggca ccgaaaaagt tcgaaacaag aagtttgttc ctgataacaa
     2161 ggatttcttt attggtggga cttcatttac tgtctccaag aaggacattt ctgctgtcat
     2221 aaccgagata gtctcacagt tcgagtctac tgatgaccaa aagtcagaga aatttaccat
     2281 gaaatctcct cctccggttg ctttagttgg gcataatctg ataggaggcc tcaagacact
     2341 gaagaatgcg ggtatcacca ttcccatact tccaatcata ggatccggcg cgccgatact
     2401 cgagaattat ggcttaatca agtgaataca tcaaagtcaa acttaaaata cattcttcgc
     2461 aggcttggct tgccacatag ttttcttcac aatgctgcaa atgacgctta ttataccctt
     2521 ttggcatcgt tgaagcttgc acaaacgaac gtcccactta atcttctgta ctctgaagag
     2581 gagtgggaaa taccaagaaa aacatcaaac tcgaatgatt ttcccgaacc cctaccacaa
     2641 gatattcatc agctgcgaga taggctgatc aggagcaagc tcgtacgaga agaaacaaaa
     2701 tgacaaaaaa aatcctatac tatataggtt acaaataaaa aagtatcaaa aatgaagcct
     2761 gcatctctca ggcaaatggc attctgacat cctct
//
