LOCUS       sequence-191917-        2753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB1 with insert: promoter PADH2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2753)
  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 2753)
  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_pADH2
FEATURES             Location/Qualifiers
     source          1..2753
                     /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(1049..1065)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1049..1065)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      1113..1144
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1145..1247
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1248..2063
                     /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(1321..1340)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2087..2114
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(2126..2713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2206..2225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(2753,1..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 cgcggagtga ttatgtaaga agaggggggt gattcggccg
      181 gctatcgaac tctaacaact aggggggtga acaatgccca gcagtcctcc ccactctttg
      241 acaaatcagt atcaccgatt aacaccccaa atcttattct caacggtccc tcatccttgc
      301 acccctcttt ggacaaatgg cagttagcat tggtgcactg actgactgcc caaccttaaa
      361 cccaaatttc ttagaagggg cccatctagt tagcgagggg tgaaaaattc ctccatcgga
      421 gatgtattga ccgtaagttg ctgcttaaaa aaaatcagtt cagatagcga gacttttttg
      481 atttcgcaac gggagtgcct gttccattcg attgcaattc tcaccccttc tgcccagtcc
      541 tgccaattgc ccatgaatct gctaatttcg ttgattccca cccccctttc caactccaca
      601 aattgtccaa tctcgttttc catttgggag aatctgcatg tcgactacat aaagcgaccg
      661 gtgtccgaaa agatctgtgt agttttcaac attttgtgct ccccccgctg tttgaaaacg
      721 ggggtgagcg ctctccgggg tgcgaattcg tgcccaattc ctttcaccct gcctattgta
      781 gacgtcaacc cgcatctggt gcgaatatag cgcaccccca atgatcacac caacaattgg
      841 tccacccctc cccaatctct aatattcaca attcacctca ctataaatac ccctgtcctg
      901 ctcccaaatt cttttttcct tcttccatca gctactagct tttatcttat ttactttacg
      961 aacatggcgt cttcgaattc gcgcgtgatc ttacggcatt atacgtatga tcggtccacg
     1021 atcagctaga ttatctagtc agcttgatgt catagctgtt tcctgaggct caatactgac
     1081 catttaaatc atacctgacc tccatagcag aaagtcaaaa gcctccgacc ggaggctttt
     1141 gacttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg
     1201 gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg agccatattc
     1261 aacgggaaac gtcttgcttg aagccgcgat taaattccaa catggatgct gatttatatg
     1321 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgattgtatg
     1381 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
     1441 ttacagatga gatggtcagg ctaaactggc tgacggaatt tatgcctctt ccgaccatca
     1501 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc ccagggaaaa
     1561 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
     1621 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacggcgatc
     1681 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt ggtgcgagtg
     1741 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
     1801 tcttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
     1861 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
     1921 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
     1981 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcact
     2041 tgatgctcga tgagtttttc taatgaggac ctaaatgtaa tcacctggct caccttcggg
     2101 tgggcctttc tgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2161 aaaatcgatg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2221 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2281 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2341 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2401 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2461 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2521 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2581 tctgcgctct gctgaagcca gttacctcgg aaaaagagtt ggtagctctt gatccggcaa
     2641 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     2701 aaaaggatct caagaagatc ctttgatttt ctaccgaaga aaggcccacc cgt
//
