LOCUS       sequence-190958-        1966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  empty BB1 for terminator cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1966)
  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 1966)
  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
FEATURES             Location/Qualifiers
     source          1..1966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     288..305
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     289..305
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(550..566)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(550..566)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      614..645
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        646..748
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             749..1564
                     /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(822..841)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1588..1615
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(join(1627..1966,1..248))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1707..1726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
       61 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
      121 gctctgctga agccagttac ctcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
      181 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
      241 gatctcaaga agatcctttg attttctacc gaagaaaggc ccacccgtgt aaaacgacgg
      301 ccagtttatc tagtcagctt gattctagct gatcgtggac cggaaggtga gccagtgagt
      361 tgattgcagt ccagttacgc tggagtctga ggctcgtcct gaatgatatg cggcctcgaa
      421 ttcgaagacg cgctttgaga ccggatccat ctcgagggtc tcacgctgcg tcttcgaatt
      481 cgcgcgtgat cttacggcat tatacgtatg atcggtccac gatcagctag attatctagt
      541 cagcttgatg tcatagctgt ttcctgaggc tcaatactga ccatttaaat catacctgac
      601 ctccatagca gaaagtcaaa agcctccgac cggaggcttt tgacttgatc ggcacgtaag
      661 aggttccaac tttcaccata atgaaataag atcactaccg ggcgtatttt ttgagttatc
      721 gagattttca ggagctaagg aagctaaaat gagccatatt caacgggaaa cgtcttgctt
      781 gaagccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
      841 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
      901 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
      961 gctaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     1021 tgatgatgca tggttactca ccactgcgat cccagggaaa acagcattcc aggtattaga
     1081 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     1141 gcattcgatt cctgtttgta attgtccttt taacggcgat cgcgtatttc gtctcgctca
     1201 ggcgcaatca cgaatgaata acggtttggt tggtgcgagt gattttgatg acgagcgtaa
     1261 tggctggcct gttgaacaag tctggaaaga aatgcataaa ctcttgccat tctcaccgga
     1321 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     1381 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     1441 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     1501 tggtattgat aatcctgata tgaataaatt gcagtttcac ttgatgctcg atgagttttt
     1561 ctaatgagga cctaaatgta atcacctggc tcaccttcgg gtgggccttt ctgcgttgct
     1621 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgat gctcaagtca
     1681 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1741 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1801 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1861 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1921 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcg
//
