LOCUS Exported 5416 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pKT0146 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5416 /organism="synthetic DNA construct" /mol_type="other DNA" 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(4027..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 //