LOCUS       sequence-192059-        2893 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  empty BB3 for insertion and overexpression of 5 genes in P. 
            pastoris.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2893)
  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 2893)
  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_AF
FEATURES             Location/Qualifiers
     source          1..2893
                     /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..1880
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        761..1104
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1105..1677
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1683..1880
                     /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 ccgcttcatg gacatggagg cccagaatac
      781 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
      841 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc
      901 gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac
      961 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
     1021 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
     1081 catcacatcc gaacataaac aaccatgggt accactcttg acgacacggc ttaccggtac
     1141 cgcaccagtg tcccggggga cgccgaggcc atcgaggcac tggatgggtc cttcaccacc
     1201 gacaccgttt ttcgcgtcac cgccaccggg gacggcttca ccctgcggga ggtgccggtg
     1261 gacccgcccc tgaccaaggt gttccccgac gacgaatcgg acgacgaatc ggacgacggg
     1321 gaggacggcg acccggactc ccggacgttc gtcgcgtacg gggacgacgg cgacctggcg
     1381 ggcttcgtgg tcgtctcgta ctccggctgg aaccgccggc tgaccgtcga ggacatcgag
     1441 gtcgccccgg agcaccgggg gcacggggtc gggcgcgcgt tgatggggct cgcgacggag
     1501 ttcgcccgcg agcggggcgc cgggcacctc tggctggagg tcaccaacgt caacgcaccg
     1561 gcgatccacg cgtaccggcg gatggggttc accctctgcg gcctggacac cgccctgtac
     1621 gacggcaccg cctcggacgg cgagcaggcg ctctacatga gcatgccctg cccctaatca
     1681 gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag tttttttata
     1741 ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt cgcctcgaca
     1801 tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc aatcgtatgt
     1861 gaatgctggt cgctatactg gcttcatacc cagcattgac ctttggtatg agcatctgaa
     1921 aaacaaccag gtgttgcaaa gttaaacatc cttctttgtt catatagaac ccactattca
     1981 tggtactccc caatcgaatt tcacattctg gttttgaaat tacacaccac gttagcttat
     2041 aagatttcat ataacttatt gatatacggt ttccattgtt cgaatagttg aggttgtatg
     2101 taattcgatt gaaggggcca tttttgtttc ctacttttcc tgggagctta tccgatgcgc
     2161 ttcaaagctg gaattgtaaa tatagagaaa aagaaggatg ttgttttatt cttgaaagag
     2221 tataatttta cttctagcaa ctctcccact tcgcttgact tcatttattt cttgggcaca
     2281 taggcgtagt aatctagacc aacagataat ttgccggaat gatatagcga ttggaaaatg
     2341 aactgaaatt ttttgctgtc tttcaatttg acgggcagtt catcaggcgc gccgtgaccg
     2401 accatataaa tacgttgaga atgttattct tcctcgtagt tgaagtggct tcataatttc
     2461 agaactcaat agataaacta ggatgtttta aagcaattaa tgctcacaag taaggagcga
     2521 ctctcttgct tttcgaatac taaaagtatc gtcccaaccc agaaaaaaag acctcttaac
     2581 tgcaaaataa actctatata tttcttctaa aacagtttca ggttggatag tatcgcattc
     2641 tcatcacttc taactagtag gccatgagat atattaacgt ttacttgagt tctaagttct
     2701 ccgaattaga tgcacagcac aaacaagatt aggtttcact tggtacaaaa tacgaacaga
     2761 gtttaaggtc gtaatttcat ttcgttattg atccccacaa tctattctta tcacagtcat
     2821 cagatagtcg cgaaaaagca tgcagaaaag ggggtcgtcc ctatctaagt tgtagcatta
     2881 caacaaatat gac
//
