LOCUS GM-pESC-URA 6495 bp DNA circular SYN 15-JUL-2025 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 6495) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 4, 2025 from SnapGene 8.1.1 https://www.snapgene.com FEATURES Location/Qualifiers source 1..6495 /mol_type="other DNA" /organism="recombinant plasmid" promoter 201..416 /gene="S. cerevisiae URA3" /label=URA3 promoter CDS 417..1220 /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" rep_origin complement(1351..1806) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1860..1884) /label=GMVec-R primer_bind 1865..1932 /label=GMIns-F promoter 1885..2528 /gene="S. cerevisiae TDH3" /label=GAP promoter /note="promoter for glyceraldehyde-3-phosphate dehydrogenase; also known as the TDH3 promoter" primer_bind 1885..1917 /label=pGAP-F misc_feature 2533..2566 /label=psiMCS /note="multiple cloning site 1" primer_bind complement(2552..2576) /label=片段 1.反向 primer_bind 2557..2596 /label=片段 2.正向 CDS join(2573..2629,2630..2827,2828..2839) /codon_start=1 /gene="MF-alpha-1" /product="N-terminal secretion signal from S. cerevisiae alpha-factor" /label=alpha-factor secretion signal /note="Cleavage by the Kex2 protease occurs after the dibasic KR sequence. The EA dipeptides are then removed by dipeptidyl aminopeptidase A." /translation="MRFPSIFTAVLFAASSALAAPVNTTTEDETAQIPAEAVIGYSDLE GDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA" primer_bind complement(2816..2859) /label=GMIns-R primer_bind 2826..2848 /label=MFM-F misc_feature 2840..2897 /label=MCS 2 /note="multiple cloning site 2" primer_bind 2840..2867 /label=GMVec-F CDS 2843..2872 /codon_start=1 /product="Myc (human c-Myc oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" terminator 2902..3091 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(3020..3038) /label=tCYC1-R rep_origin complement(3334..3922) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(join(4093..4884,4885..4953)) /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" promoter complement(4954..5058) /gene="bla" /label=AmpR promoter rep_origin 5085..6427 /label=2u ori /note="yeast 2u plasmid origin of replication" BASE COUNT 1848 a 1379 c 1462 g 1806 t ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accataccac agcttttcaa ttcaattcat catttttttt ttattctttt ttttgatttc 241 ggtttctttg aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg 301 agcacagact tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc 361 cagtattctt aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt 421 cgaaagctac atataaggaa cgtgctgcta ctcatcctag tcctgttgct gccaagctat 481 ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc attggatgtt cgtaccacca 541 aggaattact ggagttagtt gaagcattag gtcccaaaat ttgtttacta aaaacacatg 601 tggatatctt gactgatttt tccatggagg gcacagttaa gccgctaaag gcattatccg 661 ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc tgacattggt aatacagtca 721 aattgcagta ctctgcgggt gtatacagaa tagcagaatg ggcagacatt acgaatgcac 781 acggtgtggt gggcccaggt attgttagcg gtttgaagca ggcggcagaa gaagtaacaa 841 aggaacctag aggccttttg atgttagcag aattgtcatg caagggctcc ctatctactg 901 gagaatatac taagggtact gttgacattg cgaagagcga caaagatttt gttatcggct 961 ttattgctca aagagacatg ggtggaagag atgaaggtta cgattggttg attatgacac 1021 ccggtgtggg tttagatgac aagggagacg cattgggtca acagtataga accgtggatg 1081 atgtggtctc tacaggatct gacattatta ttgttggaag aggactattt gcaaagggaa 1141 gggatgctaa ggtagagggt gaacgttaca gaaaagcagg ctgggaagca tatttgagaa 1201 gatgcggcca gcaaaactaa aaaactgtat tataagtaaa tgcatgtata ctaaactcac 1261 aaattagagc ttcaatttaa ttatatcagt tattacccta tgcggtgtga aataccgcac 1321 agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt ttgttaaaat 1381 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa 1441 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca 1501 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg 1561 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta 1621 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg 1681 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa 1741 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg 1801 gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc 1861 ttcgctatta cgccagctga attgtcatta tcaatactcg ccatttcaaa gaatacgtaa 1921 ataattaata gtagtgattt tcctaacttt atttagtcaa aaaattagcc ttttaattct 1981 gctgtaaccc gtacatgccc aaaatagggg gcgggttaca cagaatatat aacatcgtag 2041 gtgtctgggt gaacagttta ttcctggcat ccactaaata taatggagcc cgctttttaa 2101 gctggcatcc agaaaaaaaa agaatcccag caccaaaata ttgttttctt caccaaccat 2161 cagttcatag gtccattctc ttagcgcaac tacagagaac aggggcacaa acaggcaaaa 2221 aacgggcaca acctcaatgg agtgatgcaa cctgcctgga gtaaatgatg acacaaggca 2281 attgacccac gcatgtatct atctcatttt cttacacctt ctattacctt ctgctctctc 2341 tgatttggaa aaagctgaaa aaaaaggttg aaaccagttc cctgaaatta ttcccctact 2401 tgactaataa gtatataaag acggtaggta ttgattgtaa ttctgtaaat ctatttctta 2461 aacttcttaa attctacttt tatagttagt ctttttttta gttttaaaac accagaactt 2521 agtttcgaat tcgaattcaa ccctcactaa agggcggccg cactaggcca ccatgagatt 2581 tccttcaatt tttactgcag ttttattcgc agcatcctcc gcattagctg ctccagtcaa 2641 cactacaaca gaagatgaaa cggcacaaat tccggctgaa gctgtcatcg gttactcaga 2701 tttagaaggg gatttcgatg ttgctgtttt gccattttcc aacagcacaa ataacgggtt 2761 attgtttata aatactacta ttgccagcat tgctgctaaa gaagaagggg tatctctcga 2821 gaaaagagag gctgaagcta tggaacagaa gttgatttcc gaagaagacc tcgagtaagc 2881 ttggtaccgc ggctagctaa gatccgctct aaccgaaaag gaaggagtta gacaacctga 2941 agtctaggtc cctatttatt tttttatagt tatgttagta ttaagaacgt tatttatatt 3001 tcaaattttt cttttttttc tgtacagacg cgtgtacgca tgtaacatta tactgaaaac 3061 cttgcttgag aaggttttgg gacgctcgaa gatccagctg cattaatgaa tcggccaacg 3121 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 3181 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 3241 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 3301 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 3361 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 3421 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 3481 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 3541 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 3601 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 3661 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 3721 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt 3781 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 3841 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 3901 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 3961 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 4021 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 4081 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 4141 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 4201 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 4261 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 4321 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 4381 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 4441 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 4501 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 4561 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 4621 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 4681 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 4741 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 4801 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 4861 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 4921 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 4981 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 5041 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgaacgaa gcatctgtgc 5101 ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga 5161 gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct 5221 gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa 5281 tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag 5341 aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa 5401 agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact 5461 ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc 5521 cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc 5581 attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact 5641 ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt 5701 tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc 5761 gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat 5821 aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg 5881 gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt 5941 ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt 6001 ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac 6061 tttctagaga ataggaactt cggaatagga acttcaaagc gtttccgaaa acgagcgctt 6121 ccgaaaatgc aacgcgagct gcgcacatac agctcactgt tcacgtcgca cctatatctg 6181 cgtgttgcct gtatatatat atacatgaga agaacggcat agtgcgtgtt tatgcttaaa 6241 tgcgtactta tatgcgtcta tttatgtagg atgaaaggta gtctagtacc tcctgtgata 6301 ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc actacccttt agctgttcta 6361 tatgctgcca ctcctcaatt ggattagtct catccttcaa tgctatcatt tcctttgata 6421 ttggatcata ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca 6481 cgaggccctt tcgtc //