LOCUS       sequence-192016-        2423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB1 with insert: RPS25Att.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2423)
  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 2423)
  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_34_RPS25Att
FEATURES             Location/Qualifiers
     source          1..2423
                     /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(719..735)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(719..735)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      783..814
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        815..917
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             918..1733
                     /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(991..1010)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1757..1784
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1796..2383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1876..1895)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(2423,1..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 cgcgctttta gtgtacatct gataatatag tactaccacg
      181 tatgataatg tagagaatag gcttccttgt cgagtgtgtt tgcagttttc ttgagtttca
      241 aggtttaaat gctggtatat tagttcatcg aaggtttcag ccaatagcac cttaaatcaa
      301 tcaaactaat tcgactctta cgaaagagcc tactgtgttt agtatcgaag tcgtttacct
      361 ttcatgttga atagcttcct ctctgaccct aacatttcaa gatcctccta aagttacccg
      421 gattgtgaaa ttctaatgat ccacctgccc aatgcatttt ttctttattc agtttacctt
      481 ttttacctaa tatacgagct tgttaaagta agtggcactg caatactagg cttattgttg
      541 atattatgat gaatcgtttt cacaaacttg atttcctgtg aactcaccat gtactaagga
      601 aaaaaacatg catcaccatc tgaatatttg accgctgcgt cttcgaattc gcgcgtgatc
      661 ttacggcatt atacgtatga tcggtccacg atcagctaga ttatctagtc agcttgatgt
      721 catagctgtt tcctgaggct caatactgac catttaaatc atacctgacc tccatagcag
      781 aaagtcaaaa gcctccgacc ggaggctttt gacttgatcg gcacgtaaga ggttccaact
      841 ttcaccataa tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag
      901 gagctaagga agctaaaatg agccatattc aacgggaaac gtcttgcttg aagccgcgat
      961 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     1021 aatcaggtgc gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga
     1081 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcagg ctaaactggc
     1141 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     1201 ggttactcac cactgcgatc ccagggaaaa cagcattcca ggtattagaa gaatatcctg
     1261 attcaggtga aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc
     1321 ctgtttgtaa ttgtcctttt aacggcgatc gcgtatttcg tctcgctcag gcgcaatcac
     1381 gaatgaataa cggtttggtt ggtgcgagtg attttgatga cgagcgtaat ggctggcctg
     1441 ttgaacaagt ctggaaagaa atgcataaac tcttgccatt ctcaccggat tcagtcgtca
     1501 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     1561 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     1621 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     1681 atcctgatat gaataaattg cagtttcact tgatgctcga tgagtttttc taatgaggac
     1741 ctaaatgtaa tcacctggct caccttcggg tgggcctttc tgcgttgctg gcgtttttcc
     1801 ataggctccg cccccctgac gagcatcaca aaaatcgatg ctcaagtcag aggtggcgaa
     1861 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1921 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1981 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2041 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2101 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2161 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2221 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttacctcgg
     2281 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2341 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatttt
     2401 ctaccgaaga aaggcccacc cgt
//
