LOCUS Exported 4894 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pKT0150 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4894 /organism="synthetic DNA construct" /mol_type="other DNA" 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(3799..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(4888..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 //