LOCUS       sequence-85544-e        2169 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FLAG-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2169)
  AUTHORS   Radeck J, Kraft K, Bartels J, Cikovic T, Durr F, Emenegger J, 
            Kelterborn S, Sauer C, Fritz G, Gebhard S, Mascher T
  TITLE     The Bacillus BioBrick Box: generation and evaluation of essential 
            genetic building blocks for standardized work with Bacillus 
            subtilis.
  JOURNAL   J Biol Eng. 2013 Dec 2;7(1):29. doi: 10.1186/1754-1611-7-29.
  PUBMED    24295448
REFERENCE   2  (bases 1 to 2169)
  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: "J Biol 
            Eng."; date: "2013-12-2"; volume: "7(1):29. doi"; pages: " 
            10.1186/1754-1611-7-29"
COMMENT     Sequence Label: FLAG-tag
FEATURES             Location/Qualifiers
     source          1..2169
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             46..111
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    121..141
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      142..199
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(223..240)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(394..982)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(474..493)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1165..1259)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1280..1939)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1854..1873
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2101..2119
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2123..2166)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
ORIGIN
        1 aattccgcgg ccgcttctag ataaggagga actactatgg ccggcgatta taaggatcat
       61 gatggtgatt ataaggatca tgatatcgac tacaaagacg atgacgacaa gaccggttaa
      121 tactagtagc ggccgctgca gtccggcaaa aaagggcaag gtgtcaccac cctgcccttt
      181 ttctttaaaa ccgaaaagat tacttcgcgt tatgcaggct tcctcgctca ctgactcgct
      241 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
      301 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
      361 caggaaccgt aaaaaggccg cgttgctggc gtttttccac aggctccgcc cccctgacga
      421 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
      481 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
      541 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
      601 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
      661 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
      721 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
      781 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
      841 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
      901 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
      961 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1021 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1081 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1141 ttggtctgac agctcgaggc ttggattctc accaataaaa aacgcccggc ggcaaccgag
     1201 cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca acaggagtcc
     1261 aagcgagctc gatatcaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt
     1321 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     1381 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     1441 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     1501 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     1561 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     1621 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     1681 tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga tgagcattca
     1741 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     1801 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     1861 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     1921 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa
     1981 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgcccga
     2041 tcaactcgag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     2101 aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat
     2161 gatttctgg
//
