LOCUS       sequence-177321-        1966 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Linker to construct BB1 from PCR product with FS2 and FS3 (in with 
            BsaI, out with BbsI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1966)
  AUTHORS   Sarkari P, Marx H, Blumhoff ML, Mattanovich D, Sauer M, Steiger MG
  TITLE     An efficient tool for metabolic pathway construction and gene 
            integration for Aspergillus niger.
  JOURNAL   Bioresour Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 
            10.1016/j.biortech.2017.05.004.
  PUBMED    28533066
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: "Bioresour 
            Technol. 2017 May 4. pii: S0960-8524(17)30643-0. doi: 
            10.1016/j.biortech.2017.05.004."
COMMENT     Sequence Label: BB1_L_23_syn_BsaI
FEATURES             Location/Qualifiers
     source          1..1966
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(17..33)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(17..33)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      81..112
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        113..215
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             216..1031
                     /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(289..308)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1055..1082
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1094..1681)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1174..1193)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1721..1738
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1722..1738
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 atctagtcag cttgatgtca tagctgtttc ctgaggctca atactgacca tttaaatcat
       61 acctgacctc catagcagaa agtcaaaagc ctccgaccgg aggcttttga cttgatcggc
      121 acgtaagagg ttccaacttt caccataatg aaataagatc actaccgggc gtattttttg
      181 agttatcgag attttcagga gctaaggaag ctaaaatgag ccatattcaa cgggaaacgt
      241 cttgcttgaa gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
      301 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
      361 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
      421 tggtcaggct aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
      481 gtactcctga tgatgcatgg ttactcacca ctgcgatccc agggaaaaca gcattccagg
      541 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
      601 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cggcgatcgc gtatttcgtc
      661 tcgctcaggc gcaatcacga atgaataacg gtttggttgg tgcgagtgat tttgatgacg
      721 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactc ttgccattct
      781 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
      841 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
      901 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
      961 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcacttg atgctcgatg
     1021 agtttttcta atgaggacct aaatgtaatc acctggctca ccttcgggtg ggcctttctg
     1081 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgatgct
     1141 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1201 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1261 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1321 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1381 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1441 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1501 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1561 tgaagccagt tacctcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1621 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1681 agaagatcct ttgattttct accgaagaaa ggcccacccg tgtaaaacga cggccagttt
     1741 atctagtcag cttgattcta gctgatcgtg gaccggaagg tgagccagtg agttgattgc
     1801 agtccagtta cgctggagtc tgaggctcgt cctgaatgat atgcggcctc gaattcgaag
     1861 acgccatgtg agaccggatc catctcgagg gtctcagctt gcgtcttcga attcgcgcgt
     1921 gatcttacgg cattatacgt atgatcggtc cacgatcagc tagatt
//
