LOCUS       sequence-190959-        2429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BB1 with insert: promoter PGAP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2429)
  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 2429)
  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_pGAP
FEATURES             Location/Qualifiers
     source          1..2429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(37..56)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     584..601
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     585..601
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        738..1214
                     /label=GAP promoter
                     /note="promoter for Pichia pastoris 
                     glyceraldehyde-3-phosphate dehydrogenase"
     primer_bind     complement(1309..1325)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1309..1325)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      1373..1404
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        1405..1507
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1508..2323
                     /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(1581..1600)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2347..2374
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(join(2386..2429,1..544))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
       61 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      121 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      181 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      241 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      301 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      361 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      421 tgctgaagcc agttacctcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      481 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      541 tcaagaagat cctttgattt tctaccgaag aaaggcccac ccgtgtaaaa cgacggccag
      601 tttatctagt cagcttgatt ctagctgatc gtggaccgga aggtgagcca gtgagttgat
      661 tgcagtccag ttacgctgga gtctgaggct cgtcctgaat gatatgcggc ctcgaattcg
      721 aagacgcgga ggatcctttt ttgtagaaat gtcttggtgt cctcgtccaa tcaggtagcc
      781 atctctgaaa tatctggctc cgttgcaact ccgaacgacc tgctggcaac gtaaaattct
      841 ccggggtaaa acttaaatgt ggagtaatgg aaccagaaac gtctcttccc ttctctctcc
      901 ttccaccgcc cgttaccgtc cctaggaaat tttactctgc tggagagctt cttctacggc
      961 ccccttgcag caatgctctt cccagcatta cgttgcgggt aaaacggagg tcgtgtaccc
     1021 gacctagcag cccagggatg gaaaagtccc ggccgtcgct ggcaataata gcgggcggac
     1081 gcatgtcatg agattattgg aaaccaccag aatcgaatat aaaaggcgaa cacctttccc
     1141 aattttggtt tctcctgacc caaagacttt aaatttaatt tatttgtccc tatttcaatc
     1201 aattgaacaa ctatcaaaac accatggcgt cttcgaattc gcgcgtgatc ttacggcatt
     1261 atacgtatga tcggtccacg atcagctaga ttatctagtc agcttgatgt catagctgtt
     1321 tcctgaggct caatactgac catttaaatc atacctgacc tccatagcag aaagtcaaaa
     1381 gcctccgacc ggaggctttt gacttgatcg gcacgtaaga ggttccaact ttcaccataa
     1441 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
     1501 agctaaaatg agccatattc aacgggaaac gtcttgcttg aagccgcgat taaattccaa
     1561 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc
     1621 gacaatctat cgattgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa
     1681 aggtagcgtt gccaatgatg ttacagatga gatggtcagg ctaaactggc tgacggaatt
     1741 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac
     1801 cactgcgatc ccagggaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga
     1861 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa
     1921 ttgtcctttt aacggcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa
     1981 cggtttggtt ggtgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt
     2041 ctggaaagaa atgcataaac tcttgccatt ctcaccggat tcagtcgtca ctcatggtga
     2101 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg
     2161 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga
     2221 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat
     2281 gaataaattg cagtttcact tgatgctcga tgagtttttc taatgaggac ctaaatgtaa
     2341 tcacctggct caccttcggg tgggcctttc tgcgttgctg gcgtttttcc ataggctccg
     2401 cccccctgac gagcatcaca aaaatcgat
//
