LOCUS       sequence-190955-        1966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  empty BB1 for promoter 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_12
FEATURES             Location/Qualifiers
     source          1..1966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     391..408
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     392..408
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(653..669)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(653..669)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      717..748
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        749..851
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             852..1667
                     /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(925..944)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1691..1718
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(join(1730..1966,1..351))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1810..1829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
       61 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      121 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      181 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      241 tacctcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      301 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      361 ttgattttct accgaagaaa ggcccacccg tgtaaaacga cggccagttt atctagtcag
      421 cttgattcta gctgatcgtg gaccggaagg tgagccagtg agttgattgc agtccagtta
      481 cgctggagtc tgaggctcgt cctgaatgat atgcggcctc gaattcgaag acgcggagtg
      541 agaccggatc catctcgagg gtctcacatg gcgtcttcga attcgcgcgt gatcttacgg
      601 cattatacgt atgatcggtc cacgatcagc tagattatct agtcagcttg atgtcatagc
      661 tgtttcctga ggctcaatac tgaccattta aatcatacct gacctccata gcagaaagtc
      721 aaaagcctcc gaccggaggc ttttgacttg atcggcacgt aagaggttcc aactttcacc
      781 ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta
      841 aggaagctaa aatgagccat attcaacggg aaacgtcttg cttgaagccg cgattaaatt
      901 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
      961 gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     1021 gcaaaggtag cgttgccaat gatgttacag atgagatggt caggctaaac tggctgacgg
     1081 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     1141 tcaccactgc gatcccaggg aaaacagcat tccaggtatt agaagaatat cctgattcag
     1201 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     1261 gtaattgtcc ttttaacggc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     1321 ataacggttt ggttggtgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     1381 aagtctggaa agaaatgcat aaactcttgc cattctcacc ggattcagtc gtcactcatg
     1441 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     1501 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     1561 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     1621 atatgaataa attgcagttt cacttgatgc tcgatgagtt tttctaatga ggacctaaat
     1681 gtaatcacct ggctcacctt cgggtgggcc tttctgcgtt gctggcgttt ttccataggc
     1741 tccgcccccc tgacgagcat cacaaaaatc gatgctcaag tcagaggtgg cgaaacccga
     1801 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1861 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1921 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcg
//
