LOCUS       sequence-160060-        4662 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Useful for tagging yeast proteins with N-terminal CFP tag; G418 
            selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4662)
  AUTHORS   Hailey DW, Davis TN, Muller EG
  TITLE     Fluorescence resonance energy transfer using color variants of green
            fluorescent protein.
  JOURNAL   Methods Enzymol. 2002;351:34-49.
  PUBMED    12073355
REFERENCE   2  (bases 1 to 4662)
  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: "Methods 
            Enzymol."; date: "2002"; volume: "351"; pages: "34-49"
COMMENT     Sequence Label: pBS5
FEATURES             Location/Qualifiers
     source          1..4662
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(19..52)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     gene            102..1458
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        102..445
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             446..1255
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(513..532)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1261..1458
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    complement(1511..1544)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1551..2264
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     HFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2299..2318)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2300..2318)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2405..2422)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2576..3164)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2656..2675)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3335..4126,4127..4195))
                     /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     3958..3977
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4196..4300)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4368..4386
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4424..4446)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4546..4565
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        join(4646..4662,1..2)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     join(4646..4662,1)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 gaacgcggcc gccaggggat aacttcgtat agcatacatt atacgaagtt atctagagat
       61 ctgtttagct tgcctcgtcc ccgccgggtc acccggccag cgacatggag gcccagaata
      121 ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt cgcccgtaca
      181 tttagcccat acatccccat gtataatcat ttgcatccat acattttgat ggccgcacgg
      241 cgcgaagcaa aaattacggc tcctcgctgc agacctgcga gcagggaaac gctcccctca
      301 cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa ggttaggatt
      361 tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga tacagttctc
      421 acatcacatc cgaacataaa caaccatggg taaggaaaag actcacgttt cgaggccgcg
      481 attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg ataatgtcgg
      541 gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag agttgtttct
      601 gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca gactaaactg
      661 gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc ctgatgatgc
      721 atggttactc accactgcga tccccggcaa aacagcattc caggtattag aagaatatcc
      781 tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat
      841 tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc
      901 acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta atggctggcc
      961 tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg attcagtcgt
     1021 cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat taataggttg
     1081 tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca tcctatggaa
     1141 ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat atggtattga
     1201 taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt tctaatcagt
     1261 actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt
     1321 gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc
     1381 atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga
     1441 atgctggtcg ctatactgct gtcgattcga tactaacgcc gccatccagt gtcgaaaacg
     1501 agctctcgag ataacttcgt atagcataca ttatacgaag ttatcagatc atgtctaaag
     1561 gtgaagaatt attcactggt gttgtcccaa ttttggttga attagatggt gatgttaatg
     1621 gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc tacttacggt aaattgacct
     1681 taaaatttat ttgtactact ggtaaattgc cagttccatg gccaacctta gtcactactt
     1741 tgacttgggg tgttcaatgt tttgctagat acccagatca tatgaaacaa catgactttt
     1801 tcaagtctgc catgccagaa ggttatgttc aagaaagaac tatttttttc aaagatgacg
     1861 gtaactacaa gaccagagct gaagtcaagt ttgaaggtga taccttagtt aatagaatcg
     1921 aattaaaagg tattgatttt aaagaagatg gtaacatttt aggtcacaaa ttggaataca
     1981 actatatttc tcacaatgtt tacatcactg ctgacaaaca aaagaatggt atcaaagctc
     2041 atttcaaaat tagacacaac attgaagatg gttctgttca attagctgac cattatcaac
     2101 aaaatactcc aattggtgat ggtccagtct tgttaccaga caaccattac ttatccactc
     2161 aatctgcctt atccaaagat ccaaacgaaa agagagacca catggtcttg ttagaatttg
     2221 ttactgctgc tggtattacc catggtatgg atgaattgta caaactagtg gcctatgcgg
     2281 ccgcggatct gccggtctcc ctatagtgag tcgtattaat ttcgataagc caggttaacc
     2341 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2401 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2461 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2521 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2581 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2641 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     2701 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     2761 cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     2821 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     2881 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2941 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3001 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3061 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3121 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3181 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3241 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3301 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3361 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3421 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3481 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3541 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3601 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3661 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     3721 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     3781 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     3841 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3901 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3961 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4021 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4081 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4141 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4201 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4261 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4321 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     4381 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     4441 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     4501 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     4561 ttgtactgag agtgcaccat atggacatat tgtcgttaga acgcggctac aattaataca
     4621 taaccttatg tatcatacac atacgattta ggtgacacta ta
//
