LOCUS       sequence-6748-ex        5444 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5444)
  AUTHORS   Sheff MA, Thorn KS
  TITLE     Optimized cassettes for fluorescent protein tagging in Saccharomyces
            cerevisiae.
  JOURNAL   Yeast 2004 Jun;21(8):661-70.
  PUBMED    15197731
REFERENCE   2  (bases 1 to 5444)
  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: "Yeast"; 
            date: "2004-06"; volume: "21(8)"; pages: "661-70"
COMMENT     Sequence Label: pKT0233
FEATURES             Location/Qualifiers
     source          1..5444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      26..213
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(81..105)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            288..1644
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        288..631
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             632..1441
                     /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(699..718)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1447..1644
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1748..1767)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1749..1767)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1854..1871)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2025..2613)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2105..2124)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2784..3575,3576..3644))
                     /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     3407..3426
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3645..3749)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3817..3835
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3873..3895)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3995..4014
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4095..4113
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4095..4112
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4183..4896
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             4915..5412
                     /codon_start=1
                     /product="13 tandem Myc epitope tags"
                     /label=13xMyc
                     /translation="EQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLIS
                     EEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLD
                     GEQKLISEEDLNGEQKLISEEDLNGEQKLISEEDLNGLDGEQKLISEEDLNGEQKLISE
                     EDL"
ORIGIN
        1 aattcggcgc gccacttcta aataagcgaa tttcttatga tttatgattt ttattattaa
       61 ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg
      121 aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagta
      181 tgaggtcgct cttattgacc acacctctac cggcagatcc gctagggata acagggtaat
      241 atagatctgt ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac atggaggccc
      301 agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt gactgtcgcc
      361 cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat tttgatggcc
      421 gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag ggaaacgctc
      481 ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat ataaaaggtt
      541 aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg ctaggataca
      601 gttctcacat cacatccgaa cataaacaac catgggtaag gaaaagactc acgtttcgag
      661 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
      721 tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg cgccagagtt
      781 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
      841 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
      901 tgatgcatgg ttactcacca ctgcgatccc cggcaaaaca gcattccagg tattagaaga
      961 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     1021 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc
     1081 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     1141 ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct caccggattc
     1201 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     1261 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     1321 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     1381 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     1441 atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt atagtttttt
     1501 tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt ttttcgcctc
     1561 gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg cgtcaatcgt
     1621 atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca tccagtttaa
     1681 acgagctcga attcatcgat gatatcagat ccactagtgg cctatgcggc cgcggatctg
     1741 ccggtctccc tatagtgagt cgtattaatt tcgataagcc aggttaacct gcattaatga
     1801 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1861 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1921 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1981 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2041 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2101 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2161 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     2221 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2281 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2341 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2401 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2461 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2521 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2581 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2641 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2701 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2761 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2821 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2881 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2941 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3001 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3061 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3121 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3181 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3241 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3301 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3361 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3421 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3481 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3541 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3601 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3661 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3721 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3781 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     3841 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     3901 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     3961 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     4021 gtgcaccata tggacatatt gtcgttagaa cgcggctaca attaatacat aaccttatgt
     4081 atcatacaca tacgatttag gtgacactat agaacgcggc cgccagctga agcttcgtac
     4141 gctgcaggtc gacggatcgg tgacggtgct ggtttaatta acatgtctaa aggtgaagaa
     4201 ttattcactg gtgttgtccc aattttggtt gaattagatg gtgatgttaa tggtcacaaa
     4261 ttttctgtct ccggtgaagg tgaaggtgat gctacttacg gtaaattgac cttaaaattt
     4321 atttgtacta ctggtaaatt gccagttcca tggccaacct tagtcactac tttaggttat
     4381 ggtttgatgt gttttgctag atacccagat catatgaaac aacatgactt tttcaagtct
     4441 gccatgccag aaggttatgt tcaagaaaga actatttttt tcaaagatga cggtaactac
     4501 aagaccagag ctgaagtcaa gtttgaaggt gataccttag ttaatagaat cgaattaaaa
     4561 ggtattgatt ttaaagaaga tggtaacatt ttaggtcaca aattggaata caactataac
     4621 tctcacaatg tttacatcat ggctgacaaa caaaagaatg gtatcaaagt taacttcaaa
     4681 attagacaca acattgaaga tggttctgtt caattagctg accattatca acaaaatact
     4741 ccaattggtg atggtccagt cttgttacca gacaaccatt acttatccta tcaatctgcc
     4801 ttatccaaag atccaaacga aaagagagac cacatggtct tgttagaatt tgttactgct
     4861 gctggtatta cccatggtat ggatgaattg tacaaacccg ggttaattaa cggtgaacaa
     4921 aagctaatct ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg
     4981 aacggactcg acggtgaaca aaagttgatt tctgaagaag atttgaacgg tgaacaaaag
     5041 ctaatctccg aggaagactt gaacggtgaa caaaaattaa tctcagaaga agacttgaac
     5101 ggactcgacg gtgaacaaaa gttgatttct gaagaagatt tgaacggtga acaaaagcta
     5161 atctccgagg aagacttgaa cggtgaacaa aaattaatct cagaagaaga cttgaacgga
     5221 ctcgacggtg aacaaaagtt gatttctgaa gaagatttga acggtgaaca aaagctaatc
     5281 tccgaggaag acttgaacgg tgaacaaaaa ttaatctcag aagaagactt gaacggactc
     5341 gacggtgaac aaaagttgat ttctgaagaa gatttgaacg gtgaacaaaa gctaatctcc
     5401 gaggaagact tgaacggtga acaaaaatta atcaatcact agtg
//
