LOCUS       sequence-191940-        2162 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB1 with insert: promoter PPpPfk.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2162)
  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 2162)
  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: BB1_12_pPFK300
FEATURES             Location/Qualifiers
     source          1..2162
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(458..474)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(458..474)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      522..553
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        554..656
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             657..1472
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(730..749)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1496..1523
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1535..2122)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1615..1634)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(2162,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagttta tctagtcagc ttgattctag ctgatcgtgg accggaaggt
       61 gagccagtga gttgattgca gtccagttac gctggagtct gaggctcgtc ctgaatgata
      121 tgcggcctcg aattcgaaga cgcggagcct gccattaggc aaacaccaac taggaaccta
      181 caccaagtta tgaaggagct tggaaaggat tataaaaaat gcgattacct aatacacaca
      241 cgatgcctga gcggttaatg tgaaatttcc cggccatcga tcgccccaca atcagactat
      301 tttcgctcat acaataaaaa gtctcctgtt tccacccgag tatcaccaga acaaagtaac
      361 aaatacaaaa ccatggcgtc ttcgaattcg cgcgtgatct tacggcatta tacgtatgat
      421 cggtccacga tcagctagat tatctagtca gcttgatgtc atagctgttt cctgaggctc
      481 aatactgacc atttaaatca tacctgacct ccatagcaga aagtcaaaag cctccgaccg
      541 gaggcttttg acttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat
      601 cactaccggg cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatga
      661 gccatattca acgggaaacg tcttgcttga agccgcgatt aaattccaac atggatgctg
      721 atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc
      781 gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg
      841 ccaatgatgt tacagatgag atggtcaggc taaactggct gacggaattt atgcctcttc
      901 cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc
      961 cagggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg
     1021 atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta
     1081 acggcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg
     1141 gtgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa
     1201 tgcataaact cttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg
     1261 ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa
     1321 tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt
     1381 cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc
     1441 agtttcactt gatgctcgat gagtttttct aatgaggacc taaatgtaat cacctggctc
     1501 accttcgggt gggcctttct gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     1561 agcatcacaa aaatcgatgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     1621 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     1681 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     1741 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     1801 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     1861 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1921 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     1981 tatttggtat ctgcgctctg ctgaagccag ttacctcgga aaaagagttg gtagctcttg
     2041 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     2101 gcgcagaaaa aaaggatctc aagaagatcc tttgattttc taccgaagaa aggcccaccc
     2161 gt
//
