LOCUS       sequence-492176-        3260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 Flp recombinase excisable antibiotic resistance 
            cassette in the grammar of an assembled transcription unit. May be 
            omega-assembled with another transcription unit..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3260)
  AUTHORS   Cooke MB, Welch KT, Deus Ramirez L, Wen AX, Marciano DC,, Herman C
  TITLE     GoldenBraid2.0 E. coli: A comprehensive and characterized toolkit 
            for enterics.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3260)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBSKa2_FRTcamFRT
FEATURES             Location/Qualifiers
     source          1..3260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    44..77
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     promoter        99..201
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             202..861
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(268..287)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     protein_bind    874..921
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     rep_origin      complement(1060..1648)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1140..1159)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1763..1865
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1866..2681
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             1866..2681
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1939..1958)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2795..3250
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2882..2901)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3092..3113
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgtctcagtc aggagattcc ggggatccgt cgacctgcag ttcgaagttc ctattctcta
       61 gaaagtatag gaacttcaga gcgcttttga agctcacgtg atcggcacgt aagaggttcc
      121 aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt atcgagattt
      181 tcaggagcta aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata
      241 tcccaatggc atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat
      301 aaccagaccg ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac
      361 aagttttatc cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc
      421 cgtatggcaa tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc
      481 gttttccatg agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc
      541 cggcagtttc tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat
      601 ttccctaaag ggtttattga gaatatgttt ttcgtgtcag ccaatccctg ggtgagtttc
      661 accagttttg atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg
      721 ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat
      781 gccgtttgtg atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat
      841 gagtggcagg gcggggcgta ataaggggat cttgaagttc ctattccgaa gttcctattc
      901 tctagaaagt ataggaactt cgaagcagct ccagcctaca cgcttgagac gaagcttgac
      961 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1021 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1081 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1141 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1201 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1261 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1321 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1381 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1441 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1501 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1561 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1621 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1681 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     1741 cttcacctag atccttttgt gatgatcggc acgtaagagg ttccaacttt caccataatg
     1801 aaataagatc actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag
     1861 ctaaaatgag ccatattcaa cgggaaacgt cttgctcaag gccgcgatta aattccaaca
     1921 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     1981 caatctaccg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     2041 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     2101 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     2161 ctgcgatccc agggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     2221 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     2281 gtccttttaa cagcgatcgc gtatttcgac tcgctcaggc gcaatcacga atgaataacg
     2341 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     2401 ggaaagaaat gcataaactt ttgccattct caccggattc agtagtcact catggtgatt
     2461 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2521 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     2581 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     2641 ataaattgca gtttcatttg atgctcgatg agtttttctg atttttttta aggcagttat
     2701 tggtgccctt aaacgcctgg ttgctacgcc tgaataagtg ataataagcg gatgaatggc
     2761 agaaattcga tgataagctg tcaaacatga tgttacgcgc cctgtagcgg cgcattaagc
     2821 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2881 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2941 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3001 aaacttgatt agggtgatgg ttcacgtagt gggccagcgc cctgatagac ggtttttcgc
     3061 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3121 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3181 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3241 cttacaattt taacaagctt
//
