LOCUS       sequence-488059-        3358 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 3358)
  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  JOURNAL   Unpublished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 3358)
  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            "Unpublished"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: pBSKa1_FRTspcFRT
FEATURES             Location/Qualifiers
     source          1..3358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    40..73
                     /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        93..164
                     /gene="bla"
                     /label=AmpR promoter
     CDS             165..956
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     protein_bind    969..1016
                     /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(1159..1747)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1239..1258)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1862..1964
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1965..2780
                     /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             1965..2780
                     /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(2038..2057)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2894..3349
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2981..3000)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3191..3212
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgtctcagga gattccgggg atccgtcgac ctgcagttcg aagttcctat tctctagaaa
       61 gtataggaac ttcagagcgc ttttgaagct cacgttcaaa tatgtatccg ctcatgagac
      121 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagg gaagcggtga
      181 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
      241 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
      301 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
      361 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
      421 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
      481 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
      541 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
      601 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
      661 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
      721 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
      781 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
      841 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
      901 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataataag
      961 gggatcttga agttcctatt ccgaagttcc tattctctag aaagtatagg aacttcgaag
     1021 cagctccagc ctacacgctg tcatgagacg aattctgaca cggttatcca cagaatcagg
     1081 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1141 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1201 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1261 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1321 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1381 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1441 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1501 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1561 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1621 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1681 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1741 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1801 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttgtg
     1861 atgatcggca cgtaagaggt tccaactttc accataatga aataagatca ctaccgggcg
     1921 tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatgagc catattcaac
     1981 gggaaacgtc ttgctcaagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     2041 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctaccga ttgtatggga
     2101 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     2161 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     2221 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatccca gggaaaacag
     2281 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     2341 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     2401 tatttcgact cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     2461 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     2521 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     2581 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     2641 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     2701 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     2761 tgctcgatga gtttttctga ttttttttaa ggcagttatt ggtgccctta aacgcctggt
     2821 tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgat gataagctgt
     2881 caaacatgat gttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2941 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3001 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3061 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3121 tcacgtagtg ggccagcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3181 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3241 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3301 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttt aacgaatt
//
