LOCUS       sequence-192024-        2927 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  empty BB3 for insertion and overexpression of 3 genes in P. 
            pastoris.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2927)
  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 2927)
  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: BB3aK_AD
FEATURES             Location/Qualifiers
     source          1..2927
                     /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"
     gene            761..2117
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        761..1104
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1105..1914
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1172..1191)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1920..2117
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
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 accggatccc tcgagggtct ccagctcatg gacatggagg cccagaatac
      781 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
      841 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc
      901 gcgaagcaaa aattacggct cctcgctgca gacgtgcgag cagggaaacg ctcccctcac
      961 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
     1021 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
     1081 catcacatcc gaacataaac aaccatgggt aaggaaaaga ctcacgtttc gaggccgcga
     1141 ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg
     1201 caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga gttgtttctg
     1261 aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg
     1321 ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca
     1381 tggttactca ccactgcgat ccccggcaaa acagcattcc aggtattaga agaatatcct
     1441 gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt
     1501 cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca
     1561 cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct
     1621 gttgaacaag tctggaaaga aatgcataag cttttgccat tctcaccgga ttcagtcgtc
     1681 actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt
     1741 attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac
     1801 tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat
     1861 aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagta
     1921 ctgacaataa aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg
     1981 tagttgttct attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca
     2041 tctgcccaga tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa
     2101 tgctggtcgc tatactggct tcagcaacgt tgtcactgaa gttggcatca gtatctacaa
     2161 accctacata cctcttgaag gtccagaagg taagcatttg cccgctccat tcctacccga
     2221 catatccact ctacatttta tagttcaagg caccgaaaaa gttcgaaaca agaagtttgt
     2281 tcctgataac aaggatttct ttattggtgg gacttcattt actgtctcca agaaggacat
     2341 ttctgctgtc ataaccgaga tagtctcaca gttcgagtct actgatgacc aaaagtcaga
     2401 gaaatttacc atgaaatctc ctcctccggt tgctttagtt gggcataatc tgataggagg
     2461 cctcaagaca ctgaagaatg cgggtatcac cattcccata cttccaatca taggatccgg
     2521 cgcgccgata ctcgagaatt atggcttaat caagtgaata catcaaagtc aaacttaaaa
     2581 tacattcttc gcaggcttgg cttgccacat agttttcttc acaatgctgc aaatgacgct
     2641 tattataccc ttttggcatc gttgaagctt gcacaaacga acgtcccact taatcttctg
     2701 tactctgaag aggagtggga aataccaaga aaaacatcaa actcgaatga ttttcccgaa
     2761 cccctaccac aagatattca tcagctgcga gataggctga tcaggagcaa gctcgtacga
     2821 gaagaaacaa aatgacaaaa aaaatcctat actatatagg ttacaaataa aaaagtatca
     2881 aaaatgaagc ctgcatctct caggcaaatg gcattctgac atcctct
//
