LOCUS       sequence-192020-        2931 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 2931)
  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 2931)
  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_14*
FEATURES             Location/Qualifiers
     source          1..2931
                     /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            765..2121
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        765..1108
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1109..1918
                     /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(1176..1195)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1924..2121
                     /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 gcgtcttcgg atccgaagac gccgctagct catggacatg gaggcccaga
      781 ataccctcct tgacagtctt gacgtgcgca gctcaggggc atgatgtgac tgtcgcccgt
      841 acatttagcc catacatccc catgtataat catttgcatc catacatttt gatggccgca
      901 cggcgcgaag caaaaattac ggctcctcgc tgcagacgtg cgagcaggga aacgctcccc
      961 tcacagacgc gttgaattgt ccccacgccg cgcccctgta gagaaatata aaaggttagg
     1021 atttgccact gaggttcttc tttcatatac ttccttttaa aatcttgcta ggatacagtt
     1081 ctcacatcac atccgaacat aaacaaccat gggtaaggaa aagactcacg tttcgaggcc
     1141 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     1201 cgggcaatca ggtgcgacaa tctatcgatt gtatgggaag cccgatgcgc cagagttgtt
     1261 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     1321 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     1381 tgcatggtta ctcaccactg cgatccccgg caaaacagca ttccaggtat tagaagaata
     1441 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     1501 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     1561 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     1621 gcctgttgaa caagtctgga aagaaatgca taagcttttg ccattctcac cggattcagt
     1681 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     1741 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     1801 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     1861 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     1921 agtactgaca ataaaaagat tcttgttttc aagaacttgt catttgtata gtttttttat
     1981 attgtagttg ttctatttta atcaaatgtt agcgtgattt atattttttt tcgcctcgac
     2041 atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa tatcatgcgt caatcgtatg
     2101 tgaatgctgg tcgctatact ggcttcagca acgttgtcac tgaagttggc atcagtatct
     2161 acaaacccta catacctctt gaaggtccag aaggtaagca tttgcccgct ccattcctac
     2221 ccgacatatc cactctacat tttatagttc aaggcaccga aaaagttcga aacaagaagt
     2281 ttgttcctga taacaaggat ttctttattg gtgggacttc atttactgtc tccaagaagg
     2341 acatttctgc tgtcataacc gagatagtct cacagttcga gtctactgat gaccaaaagt
     2401 cagagaaatt taccatgaaa tctcctcctc cggttgcttt agttgggcat aatctgatag
     2461 gaggcctcaa gacactgaag aatgcgggta tcaccattcc catacttcca atcataggat
     2521 ccggcgcgcc gatactcgag aattatggct taatcaagtg aatacatcaa agtcaaactt
     2581 aaaatacatt cttcgcaggc ttggcttgcc acatagtttt cttcacaatg ctgcaaatga
     2641 cgcttattat acccttttgg catcgttgaa gcttgcacaa acgaacgtcc cacttaatct
     2701 tctgtactct gaagaggagt gggaaatacc aagaaaaaca tcaaactcga atgattttcc
     2761 cgaaccccta ccacaagata ttcatcagct gcgagatagg ctgatcagga gcaagctcgt
     2821 acgagaagaa acaaaatgac aaaaaaaatc ctatactata taggttacaa ataaaaaagt
     2881 atcaaaaatg aagcctgcat ctctcaggca aatggcattc tgacatcctc t
//
