LOCUS       sequence-192051-        2700 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 2700)
  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 2700)
  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: BB3aN_14*
FEATURES             Location/Qualifiers
     source          1..2700
                     /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            771..1890
                     /label=natMX6
                     /note="yeast selectable marker conferring nourseothricin 
                     resistance"
     promoter        771..1114
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1115..1687
                     /codon_start=1
                     /gene="Streptomyces noursei nat1"
                     /product="nourseothricin acetyltransferase"
                     /label=NrsR
                     /note="confers resistance to nourseothricin"
                     /translation="MGTTLDDTAYRYRTSVPGDAEAIEALDGSFTTDTVFRVTATGDGF
                     TLREVPVDPPLTKVFPDDESDDESDDGEDGDPDSRTFVAYGDDGDLAGFVVVSYSGWNR
                     RLTVEDIEVAPEHRGHGVGRALMGLATEFARERGAGHLWLEVTNVNAPAIHAYRRMGFT
                     LCGLDTALYDGTASDGEQALYMSMPCP"
     terminator      1693..1890
                     /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 gacatggagg
      781 cccagaatac cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc
      841 gcccgtacat ttagcccata catccccatg tataatcatt tgcatccata cattttgatg
      901 gccgcacggc gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg
      961 ctcccctcac agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag
     1021 gttaggattt gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat
     1081 acagttctca catcacatcc gaacataaac aaccatgggt accactcttg acgacacggc
     1141 ttaccggtac cgcaccagtg tcccggggga cgccgaggcc atcgaggcac tggatgggtc
     1201 cttcaccacc gacaccgttt ttcgcgtcac cgccaccggg gacggcttca ccctgcggga
     1261 ggtgccggtg gacccgcccc tgaccaaggt gttccccgac gacgaatcgg acgacgaatc
     1321 ggacgacggg gaggacggcg acccggactc ccggacgttc gtcgcgtacg gggacgacgg
     1381 cgacctggcg ggcttcgtgg tcgtctcgta ctccggctgg aaccgccggc tgaccgtcga
     1441 ggacatcgag gtcgccccgg agcaccgggg gcacggggtc gggcgcgcgt tgatggggct
     1501 cgcgacggag ttcgcccgcg agcggggcgc cgggcacctc tggctggagg tcaccaacgt
     1561 caacgcaccg gcgatccacg cgtaccggcg gatggggttc accctctgcg gcctggacac
     1621 cgccctgtac gacggcaccg cctcggacgg cgagcaggcg ctctacatga gcatgccctg
     1681 cccctaatca gtactgacaa taaaaagatt cttgttttca agaacttgtc atttgtatag
     1741 tttttttata ttgtagttgt tctattttaa tcaaatgtta gcgtgattta tatttttttt
     1801 cgcctcgaca tcatctgccc agatgcgaag ttaagtgcgc agaaagtaat atcatgcgtc
     1861 aatcgtatgt gaatgctggt cgctatactg gcttcagcaa cgttgtcact gaagttggca
     1921 tcagtatcta caaaccctac atacctcttg aaggtccaga aggtaagcat ttgcccgctc
     1981 cattcctacc cgacatatcc actctacatt ttatagttca aggcaccgaa aaagttcgaa
     2041 acaagaagtt tgttcctgat aacaaggatt tctttattgg tgggacttca tttactgtct
     2101 ccaagaagga catttctgct gtcataaccg agatagtctc acagttcgag tctactgatg
     2161 accaaaagtc agagaaattt accatgaaat ctcctcctcc ggttgcttta gttgggcata
     2221 atctgatagg aggcctcaag acactgaaga atgcgggtat caccattccc atacttccaa
     2281 tcataggatc cggcgcgccg atactcgaga attatggctt aatcaagtga atacatcaaa
     2341 gtcaaactta aaatacattc ttcgcaggct tggcttgcca catagttttc ttcacaatgc
     2401 tgcaaatgac gcttattata cccttttggc atcgttgaag cttgcacaaa cgaacgtccc
     2461 acttaatctt ctgtactctg aagaggagtg ggaaatacca agaaaaacat caaactcgaa
     2521 tgattttccc gaacccctac cacaagatat tcatcagctg cgagataggc tgatcaggag
     2581 caagctcgta cgagaagaaa caaaatgaca aaaaaaatcc tatactatat aggttacaaa
     2641 taaaaaagta tcaaaaatga agcctgcatc tctcaggcaa atggcattct gacatcctct
//
