LOCUS       sequence-6755-ex        5572 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 5572)
  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 5572)
  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: pKT0178
FEATURES             Location/Qualifiers
     source          1..5572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             10..687
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MVASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFL"
     CDS             730..1404
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="ASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAKL
                     KVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGVV
                     TVTQDSSLQDGTLIYKVKFRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQA
                     LKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHHL
                     FL"
     terminator      1425..1612
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(1480..1504)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            1687..3043
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        1687..2030
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2031..2840
                     /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(2098..2117)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      2846..3043
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(3147..3166)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3148..3166)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3253..3270)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3424..4012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3504..3523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4183..4974,4975..5043))
                     /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     4806..4825
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5044..5148)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5216..5234
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5272..5294)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5394..5413
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5494..5512
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5494..5511
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 ttaattaaca tggtggcctc ctccgaggac gtcatcaaag agttcatgcg cttcaaggtg
       61 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      121 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
      181 gcctgggaca tcctgtcccc ccagttccag tacggctcca aggcgtacgt gaagcacccc
      241 gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
      301 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
      361 acgctgatct acaaggtgaa gttccgcggc accaacttcc cccccgacgg ccccgtaatg
      421 cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg
      481 ctgaagggcg agatccacca ggccctgaag ctgaaggacg gcggccacta cctggtggag
      541 ttcaagacca tctacatggc caagaagccc gtgcagctgc ccggctacta ctacgtggac
      601 accaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc
      661 tccgagggcc gccaccacct gttcctgggg catggcaccg gcagcaccgg cagcggcagc
      721 tccggcaccg cctcctccga ggacgtcatc aaagagttca tgcgcttcaa ggtgcgcatg
      781 gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
      841 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
      901 gacatcctgt ccccccagtt ccagtacggc tccaaggcgt acgtgaagca ccccgccgac
      961 atccccgact acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
     1021 ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga cggcacgctg
     1081 atctacaagg tgaagttccg cggcaccaac ttcccccccg acggccccgt aatgcagaag
     1141 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag
     1201 ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag
     1261 accatctaca tggccaagaa gcccgtgcag ctgcccggct actactacgt ggacaccaag
     1321 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag
     1381 ggccgccacc acctgttcct gtagggcgcg ccacttctaa ataagcgaat ttcttatgat
     1441 ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg
     1501 actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt
     1561 tgctttctca ggtatagtat gaggtcgctc ttattgacca cacctctacc ggcagatccg
     1621 ctagggataa cagggtaata tagatctgtt tagcttgcct cgtccccgcc gggtcacccg
     1681 gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg
     1741 gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca
     1801 tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc
     1861 tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg
     1921 tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt
     1981 aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaagg
     2041 aaaagactca cgtttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     2101 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga
     2161 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     2221 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     2281 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggcaaaacag
     2341 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     2401 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     2461 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     2521 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt
     2581 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     2641 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     2701 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     2761 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     2821 tgctcgatga gtttttctaa tcagtactga caataaaaag attcttgttt tcaagaactt
     2881 gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat
     2941 ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt
     3001 aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta
     3061 acgccgccat ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc cactagtggc
     3121 ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt cgataagcca
     3181 ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3241 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3301 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3361 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3421 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3481 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3541 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3601 aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3661 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3721 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3781 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3841 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     3901 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3961 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4021 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4081 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4141 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4201 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4261 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4321 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4381 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4441 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4501 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4561 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4621 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4681 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4741 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4801 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4861 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4921 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4981 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5041 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5101 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5161 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     5221 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     5281 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     5341 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     5401 agagcagatt gtactgagag tgcaccatat ggacatattg tcgttagaac gcggctacaa
     5461 ttaatacata accttatgta tcatacacat acgatttagg tgacactata gaacgcggcc
     5521 gccagctgaa gcttcgtacg ctgcaggtcg acggatcggt gacggtgctg gt
//
