LOCUS       sequence-492173-        3489 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 3489)
  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  JOURNAL   Unpublished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 3489)
  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            "Unpublished"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_FRTampFRT
FEATURES             Location/Qualifiers
     source          1..3489
                     /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        125..229
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(230..298,299..1090)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(448..467)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     protein_bind    1103..1150
                     /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(1289..1877)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1369..1388)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1992..2094
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2095..2910
                     /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             2095..2910
                     /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(2168..2187)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3024..3479
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3111..3130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3321..3342
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgtctcagtc aggagattcc ggggatccgt cgacctgcag ttcgaagttc ctattctcta
       61 gaaagtatag gaacttcaga gcgcttttga agctcacgca ggtggcactt ttcggggaaa
      121 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      181 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      241 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      301 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
      361 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      421 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
      481 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
      541 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
      601 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
      661 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
      721 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
      781 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
      841 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
      901 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggttctc gcggtatcat
      961 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1021 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1081 gcattggtaa taaggggatc ttgaagttcc tattccgaag ttcctattct ctagaaagta
     1141 taggaacttc gaagcagctc cagcctacac gcttgagacg aagcttgaca cggttatcca
     1201 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1261 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1321 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     1381 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     1441 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1501 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1561 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1621 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1681 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1741 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1801 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1861 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1921 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1981 tccttttgtg atgatcggca cgtaagaggt tccaactttc accataatga aataagatca
     2041 ctaccgggcg tattttttga gttatcgaga ttttcaggag ctaaggaagc taaaatgagc
     2101 catattcaac gggaaacgtc ttgctcaagg ccgcgattaa attccaacat ggatgctgat
     2161 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctaccga
     2221 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     2281 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     2341 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatccca
     2401 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     2461 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     2521 agcgatcgcg tatttcgact cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     2581 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     2641 cataaacttt tgccattctc accggattca gtagtcactc atggtgattt ctcacttgat
     2701 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     2761 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     2821 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     2881 tttcatttga tgctcgatga gtttttctga ttttttttaa ggcagttatt ggtgccctta
     2941 aacgcctggt tgctacgcct gaataagtga taataagcgg atgaatggca gaaattcgat
     3001 gataagctgt caaacatgat gttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3061 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3121 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3181 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3241 gggtgatggt tcacgtagtg ggccagcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3301 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3361 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3421 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttt
     3481 aacaagctt
//
