LOCUS pESC-URA-pOpM-EGFP-Sed1 8487 bp DNA circular SYN 06-OCT-2025 DEFINITION pESC-URA-pOpM-EGFP-Sed1 ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8487) AUTHORS GenScript TITLE Direct Submission JOURNAL Exported Oct 6, 2025 from SnapGene 8.1.1 https://www.snapgene.com COMMENT http://www.genscript.com/ ORIGDB|GenBank COMMENT Sequence Label: pESC-URA-pOpM-EGFP-Sed1 FEATURES Location/Qualifiers source 1..8487 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 196..416 /gene="S. cerevisiae URA3" /label=URA3 promoter /note="color: #ffffff; direction: RIGHT" promoter 201..416 /gene="S. cerevisiae URA3" /label=URA3 promoter /note="color: #ffffff" 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)" /note="color: #993366" /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" /note="color: #ffff00" terminator complement(1885..2050) /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" /note="color: #ffffff; direction: LEFT" CDS complement(2198..2221) /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /note="color: #993366" /translation="DYKDDDDK" promoter 2268..2932 /gene="GAL1 and GAL10 genes of S. cerevisiae" /label=GAL1,10 promoter /note="divergent inducible promoter, regulated by Gal4" /note="color: #ffffff" protein_bind 2484..2601 /label=UAS /note="upstream activating sequence mediating Gal4-dependent induction" /note="color: #31849b" promoter 2943..2961 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" /note="color: #ffffff" sig_peptide 2969..3244 /codon_start=1 /label=preOst1-ProMF-alpha-1 /note="color: #ffff99" /translation="MRQVWFSWIVGLFLCFFNVSSAAPVNTTTEDETAQIPAEAVIGYS DLEGDFDVAVLPFSNSTNNGLLFINTTIASIAAKEEGVSLEKREAEA" CDS 3245..3262 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /note="color: #cc99b2" /translation="HHHHHH" CDS 3263..3973 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="color: #05fc14" /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF VTAAGITLGMDELYK" CDS 3974..4018 /codon_start=1 /label=GS linker /note="color: #a6acb3" /translation="GGGGSGGGGSGGGGS" CDS 4019..4858 /codon_start=1 /label=Sed1 /note="color: #993366" /translation="ALPTNGTSTEAPTDTTTEAPTTGLPTNGTTSAFPPTTSLPPSNTT TTPPYNPSTDYTTDYTVVTEYTTYCPEPTTFTTNGKTYTVTEPTTLTITDCPCTIEKPT TTSTTEYTVVTEYTTYCPEPTTFTTNGKTYTVTEPTTLTITDCPCTIEKPTTTSTTEYT VVTEYTTYCPEPTTFTTNGKTYTVTEPTTLTITDCPCTIEKSEAPESSVPVTESKGTTT KETGVTTKQTTANPSLTVSTVVPVSSSASSHSVVINSNGANVVVPGALGLAGVAMLFL" terminator 4894..5083 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" /note="color: #ffffff" rep_origin complement(5326..5914) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" CDS complement(6085..6945) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="color: #993366" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6946..7050) /gene="bla" /label=AmpR promoter /note="color: #ffffff; direction: LEFT" rep_origin complement(7077..8419) /direction=LEFT /label=2u ori /note="yeast 2u plasmid origin of replication" /note="color: #ffff00" protein_bind complement(8040..8087) /label=FRT /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." /note="color: #31849b; direction: LEFT" 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 ggtttccttg 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 aaattgtaag 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 attggagcga cctcatgcta tacctgagaa agcaacctga 1921 cctacaggaa agagttactc aagaataaga attttcgttt taaaacctaa gagtcacttt 1981 aaaatttgta tacacttatt ttttttataa cttatttaat aataaaaatc ataaatcata 2041 agaaattcgc ttatttagaa gtgtcaacaa cgtatctacc aacgatttga cccttttcca 2101 tcttttcgta aatttctggc aaggtagaca agccgacaac cttgattgga gacttgacca 2161 aacctctggc gaagaattgt taattaagag ctcagatctt atcgtcgtca tccttgtaat 2221 ccatcgatac tagtgcggcc gccctttagt gagggttgaa ttcgaatttt caaaaattct 2281 tacttttttt ttggatggac gcaaagaagt ttaataatca tattacatgg cattaccacc 2341 atatacatat ccatatacat atccatatct aatcttactt atatgttgtg gaaatgtaaa 2401 gagccccatt atcttagcct aaaaaaacct tctctttgga actttcagta atacgcttaa 2461 ctgctcattg ctatattgaa gtacggatta gaagccgccg agcgggtgac agccctccga 2521 aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa acgcagatgt 2581 gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt tatggttatg 2641 aagaggaaaa attggcagta acctggcccc acaaaccttc aaatgaacga atcaaattaa 2701 caaccatagg atgataatgc gattagtttt ttagccttat ttctggggta attaatcagc 2761 gaagcgatga tttttgatct attaacagat atataaatgc aaaaactgca taaccacttt 2821 aactaatact ttcaacattt tcggtttgta ttacttctta ttcaaatgta ataaaagtat 2881 caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa accccggatc 2941 cgtaatacga ctcactatag ggcccgggat gcgtcaagtc tggttcagct ggattgtcgg 3001 tttgttcttg tgtttcttca acgtttcctc tgccgctcca gttaacacca ccactgaaga 3061 tgaaactgct caaattccag ctgaagcagt cattggttac tctgacttgg aaggtgactt 3121 cgatgttgcc gtcttgccat tctctaactc caccaacaac ggtttattat tcatcaacac 3181 tactattgct tccattgctg ctaaggaaga aggtgtttct ttggaaaaac gtgaagctga 3241 agctcaccat caccaccatc actctaaggg tgaagaatta ttcaccggtg ttgtcccaat 3301 tttggttgaa ttggacggtg atgttaatgg tcacaagttt tctgtttctg gtgaaggtga 3361 aggtgacgct acctacggta aactaacgtt gaagttcatc tgtactactg gtaagttgcc 3421 agttccatgg ccaactttgg ttacgacttt gacctacggt gtccaatgtt tctccagata 3481 cccagaccac atgaagcaac acgatttctt taaatccgcc atgccagaag gttacgttca 3541 agaaagaacc atcttcttca aggatgacgg taactataag accagagctg aagttaagtt 3601 tgaaggtgat actttggtca acagaattga attaaagggt attgacttca aggaagatgg 3661 taatatcttg ggtcacaagt tggaatataa ctacaactcc cacaacgtct acatcatggc 3721 tgataaacaa aagaacggta tcaaggttaa cttcaagata agacacaaca ttgaagacgg 3781 ttccgtccaa ttggccgacc attaccaaca aaacactcca atcggcgatg gtccagtttt 3841 gttgcctgac aaccactact tgtctactca atctgctttg tctaaggacc caaacgaaaa 3901 gcgtgaccac atggttttgt tggaattcgt taccgccgct ggtatcactt taggcatgga 3961 cgaattgtac aagggtggtg gtggttccgg tggtggaggt tctggtggtg gtggttctgc 4021 tttgccaact aatggtacct ccactgaagc cccaactgac acaactaccg aagccccaac 4081 taccggtcta ccaaccaatg gtaccacttc cgctttccca ccaacgacct ctttacctcc 4141 atccaacact actacaactc caccatacaa cccatctaca gactacacta ccgattacac 4201 cgtcgtcact gaatacacca cttactgtcc agaaccaacc acttttacca ccaacggtaa 4261 gacttacacc gtcactgagc ctaccactct aactatcact gactgtccat gtactattga 4321 aaagcctaca accacttcaa ccactgaata taccgtcgta actgaatata ctacctactg 4381 tccagaacca acgaccttca ccaccaacgg taagacctat actgtcaccg aaccaaccac 4441 tcttaccatt actgattgcc catgtacgat cgaaaagcca accactactt ctaccactga 4501 atacaccgtt gttactgaat acaccaccta ctgtccagaa cctactacct tcactactaa 4561 cggtaagacc tacactgtta ctgaaccaac caccttgact atcaccgact gtccatgcac 4621 catcgaaaag tctgaagctc cagaatcttc ggttccagtt actgaatcaa agggtactac 4681 tactaaggaa accggtgtta ccaccaagca aactaccgct aacccatctt tgaccgtttc 4741 tactgtggtc ccagtttctt cttccgcttc ctctcactct gttgttatca acagcaatgg 4801 tgccaacgtt gttgttccag gtgctttggg tttggctggt gttgctatgt tgtttttgtg 4861 actcgagtaa gcttggtacc gcggctagct aagatccgct ctaaccgaaa aggaaggagt 4921 tagacaacct gaagtctagg tccctattta tttttttata gttatgttag tattaagaac 4981 gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat 5041 tatactgaaa accttgcttg agaaggtttt gggacgctcg aagatccagc tgcattaatg 5101 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct 5161 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc 5221 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg 5281 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg 5341 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg 5401 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac 5461 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca 5521 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt 5581 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc 5641 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag 5701 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac 5761 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt 5821 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa 5881 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg 5941 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa 6001 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat 6061 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc 6121 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat 6181 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc 6241 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc 6301 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag 6361 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg 6421 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg 6481 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag 6541 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt 6601 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga 6661 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc 6721 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc 6781 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc 6841 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc 6901 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca 6961 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat 7021 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgaacg 7081 aagcatctgt gcttcatttt gtagaacaaa aatgcaacgc gagagcgcta atttttcaaa 7141 caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgaaagcg ctattttacc 7201 aacgaagaat ctgtgcttca tttttgtaaa acaaaaatgc aacgcgagag cgctaatttt 7261 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga gagcgctatt 7321 ttaccaacaa agaatctata cttctttttt gttctacaaa aatgcatccc gagagcgcta 7381 tttttctaac aaagcatctt agattacttt ttttctcctt tgtgcgctct ataatgcagt 7441 ctcttgataa ctttttgcac tgtaggtccg ttaaggttag aagaaggcta ctttggtgtc 7501 tattttctct tccataaaaa aagcctgact ccacttcccg cgtttactga ttactagcga 7561 agctgcgggt gcattttttc aagataaagg catccccgat tatattctat accgatgtgg 7621 attgcgcata ctttgtgaac agaaagtgat agcgttgatg attcttcatt ggtcagaaaa 7681 ttatgaacgg tttcttctat tttgtctcta tatactacgt ataggaaatg tttacatttt 7741 cgtattgttt tcgattcact ctatgaatag ttcttactac aatttttttg tctaaagagt 7801 aatactagag ataaacataa aaaatgtaga ggtcgagttt agatgcaagt tcaaggagcg 7861 aaaggtggat gggtaggtta tatagggata tagcacagag atatatagca aagagatact 7921 tttgagcaat gtttgtggaa gcggtattcg caatatttta gtagctcgtt acagtccggt 7981 gcgtttttgg ttttttgaaa gtgcgtcttc agagcgcttt tggttttcaa aagcgctctg 8041 aagttcctat actttctaga gaataggaac ttcggaatag gaacttcaaa gcgtttccga 8101 aaacgagcgc ttccgaaaat gcaacgcgag ctgcgcacat acagctcact gttcacgtcg 8161 cacctatatc tgcgtgttgc ctgtatatat atatacatga gaagaacggc atagtgcgtg 8221 tttatgctta aatgcgtact tatatgcgtc tatttatgta ggatgaaagg tagtctagta 8281 cctcctgtga tattatccca ttccatgcgg ggtatcgtat gcttccttca gcactaccct 8341 ttagctgttc tatatgctgc cactcctcaa ttggattagt ctcatccttc aatgctatca 8401 tttcctttga tattggatca tactaagaaa ccattattat catgacatta acctataaaa 8461 ataggcgtat cacgaggccc tttcgtc //