LOCUS       sequence-207097-        4894 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 4894)
  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 4894)
  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: pKT0150
FEATURES             Location/Qualifiers
     source          1..4894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..135
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        216..234
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     216..233
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             304..1020
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSIQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGIDELYK"
     terminator      1041..1228
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1096..1120)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1303..2659
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1303..1646
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             1647..2456
                     /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(1714..1733)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2462..2659
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2763..2782)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2764..2782)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2869..2886)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3040..3628)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3120..3139)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3799..4590,4591..4659))
                     /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     4422..4441
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4660..4764)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4832..4850
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(4888..4894,1..16))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
       61 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
      121 tcagagcaga ttgtactgag agtgcaccat atggacatat tgtcgttaga acgcggctac
      181 aattaataca taaccttatg tatcatacac atacgattta ggtgacacta tagaacgcgg
      241 ccgccagctg aagcttcgta cgctgcaggt cgacggatcg gtgacggtgc tggtttaatt
      301 aacatgtcta aaggtgaaga attattcact ggtgttgtcc caattttggt tgaattagat
      361 ggtgatgtta atggtcacaa attttctgtc tccggtgaag gtgaaggtga tgctacttac
      421 ggtaaattga ccttaaaatt tatttgtact actggtaaat tgccagttcc atggccaacc
      481 ttagtcacta ctttttctta tggtgttcaa tgttttgcta gatacccaga tcatatgaaa
      541 caacatgact ttttcaagtc tgccatgcca gaaggttatg ttcaagaaag aactattttt
      601 ttcaaagatg acggtaacta caagaccaga gctgaagtca agtttgaagg tgatacctta
      661 gttaatagaa tcgaattaaa aggtattgat tttaaagaag atggtaacat tttaggtcac
      721 aaattggaat acaactttaa ctctcacaat gtttacatca tggctgacaa acaaaagaat
      781 ggtatcaaag ttaacttcaa aattagacac aacattgaag atggttctgt tcaattagct
      841 gaccattatc aacaaaatac tccaattggt gatggtccag tcttgttacc agacaaccat
      901 tacttatcca ttcaatctgc cttatccaaa gatccaaacg aaaagagaga ccacatggtc
      961 ttgttagaat ttgttactgc tgctggtatt acccatggta ttgatgaatt gtacaaataa
     1021 ggcgcgccac ttctaaataa gcgaatttct tatgatttat gatttttatt attaaataag
     1081 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
     1141 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg
     1201 tcgctcttat tgaccacacc tctaccggca gatccgctag ggataacagg gtaatataga
     1261 tctgtttagc ttgcctcgtc cccgccgggt cacccggcca gcgacatgga ggcccagaat
     1321 accctccttg acagtcttga cgtgcgcagc tcaggggcat gatgtgactg tcgcccgtac
     1381 atttagccca tacatcccca tgtataatca tttgcatcca tacattttga tggccgcacg
     1441 gcgcgaagca aaaattacgg ctcctcgctg cagacctgcg agcagggaaa cgctcccctc
     1501 acagacgcgt tgaattgtcc ccacgccgcg cccctgtaga gaaatataaa aggttaggat
     1561 ttgccactga ggttcttctt tcatatactt ccttttaaaa tcttgctagg atacagttct
     1621 cacatcacat ccgaacataa acaaccatgg gtaaggaaaa gactcacgtt tcgaggccgc
     1681 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     1741 ggcaatcagg tgcgacaatc tatcgattgt atgggaagcc cgatgcgcca gagttgtttc
     1801 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     1861 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     1921 catggttact caccactgcg atccccggca aaacagcatt ccaggtatta gaagaatatc
     1981 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     2041 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     2101 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     2161 ctgttgaaca agtctggaaa gaaatgcata agcttttgcc attctcaccg gattcagtcg
     2221 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2281 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2341 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2401 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     2461 tactgacaat aaaaagattc ttgttttcaa gaacttgtca tttgtatagt ttttttatat
     2521 tgtagttgtt ctattttaat caaatgttag cgtgatttat attttttttc gcctcgacat
     2581 catctgccca gatgcgaagt taagtgcgca gaaagtaata tcatgcgtca atcgtatgtg
     2641 aatgctggtc gctatactgc tgtcgattcg atactaacgc cgccatccag tttaaacgag
     2701 ctcgaattca tcgatgatat cagatccact agtggcctat gcggccgcgg atctgccggt
     2761 ctccctatag tgagtcgtat taatttcgat aagccaggtt aacctgcatt aatgaatcgg
     2821 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     2881 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     2941 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3001 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3061 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3121 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3181 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3241 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3301 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3361 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3421 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3481 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3541 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3601 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3661 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3721 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3781 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3841 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3901 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3961 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4021 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4081 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4141 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4201 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4261 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4321 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4381 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4441 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4501 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4561 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4621 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4681 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4741 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga
     4801 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     4861 cgcgcgtttc ggtgatgacg gtgaaaacct ctga
//
