LOCUS       sequence-6734-ex        4738 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 4738)
  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 4738)
  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: pKT0101
FEATURES             Location/Qualifiers
     source          1..4738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      19..206
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(74..98)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            281..1481
                     /label=HIS3MX6
                     /note="yeast selectable marker encoding the S. pombe his5 
                     gene, which corresponds to S. cerevisiae HIS3"
     promoter        281..624
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             625..1278
                     /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      1284..1481
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     primer_bind     complement(1585..1604)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1586..1604)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1691..1708)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1862..2450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1942..1961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2621..3412,3413..3481))
                     /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     3244..3263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3482..3586)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3654..3672
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3710..3732)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3832..3851
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3932..3950
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3932..3949
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4020..4736
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYIYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
ORIGIN
        1 cgcgccactt ctaaataagc gaatttctta tgatttatga tttttattat taaataagtt
       61 ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa acgaaaattc
      121 ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata gtatgaggtc
      181 gctcttattg accacacctc taccggcaga tccgctaggg ataacagggt aatatagatc
      241 tgtttagctt gcctcgtccc cgccgggtca cccggccagc gacatggagg cccagaatac
      301 cctccttgac agtcttgacg tgcgcagctc aggggcatga tgtgactgtc gcccgtacat
      361 ttagcccata catccccatg tataatcatt tgcatccata cattttgatg gccgcacggc
      421 gcgaagcaaa aattacggct cctcgctgca gacctgcgag cagggaaacg ctcccctcac
      481 agacgcgttg aattgtcccc acgccgcgcc cctgtagaga aatataaaag gttaggattt
      541 gccactgagg ttcttctttc atatacttcc ttttaaaatc ttgctaggat acagttctca
      601 catcacatcc gaacataaac aaccatgggt aggagggctt ttgtagaaag aaatacgaac
      661 gaaacgaaaa tcagcgttgc catcgctttg gacaaagctc ccttacctga agagtcgaat
      721 tttattgatg aacttataac ttccaagcat gcaaaccaaa agggagaaca agtaatccaa
      781 gtagacacgg gaattggatt cttggatcac atgtatcatg cactggctaa acatgcaggc
      841 tggagcttac gactttactc aagaggtgat ttaatcatcg atgatcatca cactgcagaa
      901 gatactgcta ttgcacttgg tattgcattc aagcaggcta tgggtaactt tgccggcgtt
      961 aaaagatttg gacatgctta ttgtccactt gacgaagctc tttctagaag cgtagttgac
     1021 ttgtcgggac ggccctatgc tgttatcgat ttgggattaa agcgtgaaaa ggttggggaa
     1081 ttgtcctgtg aaatgatccc tcacttacta tattcctttt cggtagcagc tggaattact
     1141 ttgcatgtta cctgcttata tggtagtaat gaccatcatc gtgctgaaag cgcttttaaa
     1201 tctctggctg ttgccatgcg cgcggctact agtcttactg gaagttctga agtcccaagc
     1261 acgaagggag tgttgtaaag agtactgaca ataaaaagat tcttgttttc aagaacttgt
     1321 catttgtata gtttttttat attgtagttg ttctatttta atcaaatgtt agcgtgattt
     1381 atattttttt tcgcctcgac atcatctgcc cagatgcgaa gttaagtgcg cagaaagtaa
     1441 tatcatgcgt caatcgtatg tgaatgctgg tcgctatact gctgtcgatt cgatactaac
     1501 gccgccatcc agtttaaacg agctcgaatt catcgatgat atcagatcca ctagtggcct
     1561 atgcggccgc ggatctgccg gtctccctat agtgagtcgt attaatttcg ataagccagg
     1621 ttaacctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     1681 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1741 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1801 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1861 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1921 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1981 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2041 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2101 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2161 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2221 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2281 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     2341 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2401 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2461 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2521 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2581 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2641 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2701 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2761 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2821 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2881 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2941 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3001 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3061 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3121 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3181 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3241 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3301 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3361 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3421 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3481 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3541 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3601 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     3661 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3721 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     3781 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     3841 agcagattgt actgagagtg caccatatgg acatattgtc gttagaacgc ggctacaatt
     3901 aatacataac cttatgtatc atacacatac gatttaggtg acactataga acgcggccgc
     3961 cagctgaagc ttcgtacgct gcaggtcgac ggatcggtga cggtgctggt ttaattaaca
     4021 tgtctaaagg tgaagaatta ttcactggtg ttgtcccaat tttggttgaa ttagatggtg
     4081 atgttaatgg tcacaaattt tctgtctccg gtgaaggtga aggtgatgct acttacggta
     4141 aattgacctt aaaatttatt tgtactactg gtaaattgcc agttccatgg ccaaccttag
     4201 tcactacttt aacttggggt gttcaatgtt tttctagata cccagatcat atgaaacaac
     4261 atgacttttt caagtctgcc atgccagaag gttatgttca agaaagaact atttttttca
     4321 aagatgacgg taactacaag accagagctg aagtcaagtt tgaaggtgat accttagtta
     4381 atagaatcga attaaaaggt attgatttta aagaagatgg taacatttta ggtcacaaat
     4441 tggaatacat ttataactct cacaatgttt acatcactgc tgacaaacaa aagaatggta
     4501 tcaaagctaa cttcaaaatt agacacaaca ttgaagatgg ttctgttcaa ttagctgacc
     4561 attatcaaca aaatactcca attggtgatg gtccagtctt gttaccagac aaccattact
     4621 tatccactca atctgcctta tccaaagatc caaacgaaaa gagagaccac atggtcttgt
     4681 tagaatttgt tactgctgct ggtattaccc atggtatgga tgaattgtac aaataagg
//
