LOCUS       sequence-6753-ex        5416 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 5416)
  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 5416)
  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: pKT0146
FEATURES             Location/Qualifiers
     source          1..5416
                     /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..2887
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        1687..2030
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             2031..2684
                     /codon_start=1
                     /gene="Schizosaccharomyces pombe his5"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=S. pombe his5
                     /note="yeast auxotrophic marker; corresponds to S. 
                     cerevisiae HIS3"
                     /translation="MGRRAFVERNTNETKISVAIALDKAPLPEESNFIDELITSKHANQ
                     KGEQVIQVDTGIGFLDHMYHALAKHAGWSLRLYSRGDLIIDDHHTAEDTAIALGIAFKQ
                     AMGNFAGVKRFGHAYCPLDEALSRSVVDLSGRPYAVIDLGLKREKVGELSCEMIPHLLY
                     SFSVAAGITLHVTCLYGSNDHHRAESAFKSLAVAMRAATSLTGSSEVPSTKGVL"
     terminator      2690..2887
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(2991..3010)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2992..3010)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3097..3114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3268..3856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3348..3367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4027..4818,4819..4887))
                     /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     4650..4669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4888..4992)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5060..5078
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5116..5138)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5238..5257
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5338..5356
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5338..5355
                     /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 atgggtagga
     2041 gggcttttgt agaaagaaat acgaacgaaa cgaaaatcag cgttgccatc gctttggaca
     2101 aagctccctt acctgaagag tcgaatttta ttgatgaact tataacttcc aagcatgcaa
     2161 accaaaaggg agaacaagta atccaagtag acacgggaat tggattcttg gatcacatgt
     2221 atcatgcact ggctaaacat gcaggctgga gcttacgact ttactcaaga ggtgatttaa
     2281 tcatcgatga tcatcacact gcagaagata ctgctattgc acttggtatt gcattcaagc
     2341 aggctatggg taactttgcc ggcgttaaaa gatttggaca tgcttattgt ccacttgacg
     2401 aagctctttc tagaagcgta gttgacttgt cgggacggcc ctatgctgtt atcgatttgg
     2461 gattaaagcg tgaaaaggtt ggggaattgt cctgtgaaat gatccctcac ttactatatt
     2521 ccttttcggt agcagctgga attactttgc atgttacctg cttatatggt agtaatgacc
     2581 atcatcgtgc tgaaagcgct tttaaatctc tggctgttgc catgcgcgcg gctactagtc
     2641 ttactggaag ttctgaagtc ccaagcacga agggagtgtt gtaaagagta ctgacaataa
     2701 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct
     2761 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga
     2821 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc
     2881 tatactgctg tcgattcgat actaacgccg ccatccagtt taaacgagct cgaattcatc
     2941 gatgatatca gatccactag tggcctatgc ggccgcggat ctgccggtct ccctatagtg
     3001 agtcgtatta atttcgataa gccaggttaa cctgcattaa tgaatcggcc aacgcgcggg
     3061 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3121 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3181 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     3241 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     3301 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     3361 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     3421 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     3481 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     3541 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     3601 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     3661 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     3721 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     3781 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     3841 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     3901 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3961 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4021 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4081 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4141 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4201 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     4261 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     4321 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     4381 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     4441 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     4501 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     4561 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     4621 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     4681 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     4741 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     4801 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     4861 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     4921 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4981 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5041 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     5101 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     5161 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     5221 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatggacat
     5281 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
     5341 taggtgacac tatagaacgc ggccgccagc tgaagcttcg tacgctgcag gtcgacggat
     5401 cggtgacggt gctggt
//
