LOCUS       sequence-192055-        2911 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 2911)
  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 2911)
  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: BB3rN_14*
FEATURES             Location/Qualifiers
     source          1..2911
                     /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            779..1898
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        779..1122
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1123..1695
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1701..1898
                     /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 atccctcgag gaagacgccg ctccggcatg ccggcatgga
      781 catggaggcc cagaataccc tccttgacag tcttgacgtg cgcagctcag gggcatgatg
      841 tgactgtcgc ccgtacattt agcccataca tccccatgta taatcatttg catccataca
      901 ttttgatggc cgcacggcgc gaagcaaaaa ttacggctcc tcgctgcaga cctgcgagca
      961 gggaaacgct cccctcacag acgcgttgaa ttgtccccac gccgcgcccc tgtagagaaa
     1021 tataaaaggt taggatttgc cactgaggtt cttctttcat atacttcctt ttaaaatctt
     1081 gctaggatac agttctcaca tcacatccga acataaacaa ccatgggtac cactcttgac
     1141 gacacggctt accggtaccg caccagtgtc ccgggggacg ccgaggccat cgaggcactg
     1201 gatgggtcct tcaccaccga caccgttttt cgcgtcaccg ccaccgggga cggcttcacc
     1261 ctgcgggagg tgccggtgga cccgcccctg accaaggtgt tccccgacga cgaatcggac
     1321 gacgaatcgg acgacgggga ggacggcgac ccggactccc ggacgttcgt cgcgtacggg
     1381 gacgacggcg acctggcggg cttcgtggtc gtctcgtact ccggctggaa ccgccggctg
     1441 accgtcgagg acatcgaggt cgccccggag caccgggggc acggggtcgg gcgcgcgttg
     1501 atggggctcg cgacggagtt cgcccgcgag cggggcgccg ggcacctctg gctggaggtc
     1561 accaacgtca acgcaccggc gatccacgcg taccggcgga tggggttcac cctctgcggc
     1621 ctggacaccg ccctgtacga cggcaccgcc tcggacggcg agcaggcgct ctacatgagc
     1681 atgccctgcc cctaatcagt actgacaata aaaagattct tgttttcaag aacttgtcat
     1741 ttgtatagtt tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata
     1801 ttttttttcg cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat
     1861 catgcgtcaa tcgtatgtga atgctggtcg ctatactggc ttcataccca gcattgacct
     1921 ttggtatgag catctgaaaa acaaccaggt gttgcaaagt taaacatcct tctttgttca
     1981 tatagaaccc actattcatg gtactcccca atcgaatttc acattctggt tttgaaatta
     2041 cacaccacgt tagcttataa gatttcatat aacttattga tatacggttt ccattgttcg
     2101 aatagttgag gttgtatgta attcgattga aggggccatt tttgtttcct acttttcctg
     2161 ggagcttatc cgatgcgctt caaagctgga attgtaaata tagagaaaaa gaaggatgtt
     2221 gttttattct tgaaagagta taattttact tctagcaact ctcccacttc gcttgacttc
     2281 atttatttct tgggcacata ggcgtagtaa tctagaccaa cagataattt gccggaatga
     2341 tatagcgatt ggaaaatgaa ctgaaatttt ttgctgtctt tcaatttgac gggcagttca
     2401 tcaggcgcgc cgtgaccgac catataaata cgttgagaat gttattcttc ctcgtagttg
     2461 aagtggcttc ataatttcag aactcaatag ataaactagg atgttttaaa gcaattaatg
     2521 ctcacaagta aggagcgact ctcttgcttt tcgaatacta aaagtatcgt cccaacccag
     2581 aaaaaaagac ctcttaactg caaaataaac tctatatatt tcttctaaaa cagtttcagg
     2641 ttggatagta tcgcattctc atcacttcta actagtaggc catgagatat attaacgttt
     2701 acttgagttc taagttctcc gaattagatg cacagcacaa acaagattag gtttcacttg
     2761 gtacaaaata cgaacagagt ttaaggtcgt aatttcattt cgttattgat ccccacaatc
     2821 tattcttatc acagtcatca gatagtcgcg aaaaagcatg cagaaaaggg ggtcgtccct
     2881 atctaagttg tagcattaca acaaatatga c
//
