LOCUS Exported 5542 bp ds-DNA circular SYN 10-FEB-2020 DEFINITION base genetic reporter construct for testing yeast promoter library. ACCESSION . VERSION . KEYWORDS RCP83 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5542) TITLE yeast cis-eQTL oligo reporter constructs and libraries JOURNAL Unpublished REFERENCE 2 (bases 1 to 5542) AUTHORS . TITLE Direct Submission JOURNAL Exported Monday, Feb 10, 2020 from SnapGene 5.0.1 https://www.snapgene.com FEATURES Location/Qualifiers source 1..5542 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 16..28 /label=SfiI misc_feature 29..131 /label=sharon2012_1922 misc_feature 140..239 /label=his3 CDS 240..953 /codon_start=1 /product="green fluorescent protein" /label=yEGFP /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" terminator 1031..1196 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(1086..1110) /label=pHybLex-R /note="ADH terminator, reverse primer" misc_feature 1512..2015 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" gene complement(2095..3451) /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" terminator 2095..2292 /label=TEF terminator /note="Ashbya gossypii TEF terminator" CDS complement(2298..3107) /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 3021..3040 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" promoter complement(3108..3451) /label=TEF promoter /note="Ashbya gossypii TEF promoter" terminator 3584..3615 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" promoter 3616..3718 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 3719..4579 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(3937..3956) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" CDS 4580..4771 /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" terminator 4795..4822 /label=T7Te terminator /note="phage T7 early transcription terminator" rep_origin complement(4834..5421) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4914..4933) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 5443..5472 /label=T3Te terminator /note="phage T3 early transcription terminator" misc_feature join(5521..5542,1..15) /label=PS1 ORIGIN 1 ccgctcttcc gatctggcca ttacggccaa gccggacagg cagcgacagc cctgacagac 61 acggaagact ctcctccgtc ctcgtcttca ccggtcgcgg aagactctcc tccgtgtgcc 121 tacagccgca cggcgcgcca ttggcattat cacataatga attatacatt atataaagta 181 atgtgatttc ttcgaagaat atactaaaaa atgagcaggc aagataaacg aaggcaaaga 241 tgtctaaagg tgaagaatta ttcactggtg ttgtcccaat tttggttgaa ttagatggtg 301 atgttaatgg tcacaaattt tctgtctccg gtgaaggtga aggtgatgct acttacggta 361 aattgacctt aaaatttatt tgtactactg gtaaattgcc agttccatgg ccaaccttag 421 tcactacttt aacttatggt gttcaatgtt tttctagata cccagatcat atgaaacaac 481 atgacttttt caagtctgcc atgccagaag gttatgttca agaaagaact atttttttca 541 aagatgacgg taactacaag accagagctg aagtcaagtt tgaaggtgat accttagtta 601 atagaatcga attaaaaggt attgatttta aagaagatgg taacatttta ggtcacaaat 661 tggaatacaa ctataactct cacaatgttt acatcatggc tgacaaacaa aagaatggta 721 tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg ttctgttcaa ttagctgacc 781 attatcaaca aaatactcca attggtgatg gtccagtctt gttaccagac aaccattact 841 tatccactca atctgcctta tccaaagatc caaacgaaaa gagagaccac atggtcttgt 901 tagaatttgt tactgctgct ggtattaccc atggtatgga tgaattgtac aaataacctg 961 caggctacag catgggagat acttagatcg gaagagcgtc gtgtagggaa agagtgtggc 1021 ctaattggcc gcgaatttct tatgatttat gatttttatt attaaataag ttataaaaaa 1081 aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat tcttattctt 1141 gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagtatgagg tcgctcttat 1201 tgaccacacc acagaagacg ggagacacta gcacacaact ttaccaggca aggtatttga 1261 cgctagcatg tgtccaattc agtgtcattt atgatttttt gtagtaggat ataaatatat 1321 acagcgctcc aaatagtgcg gttgccccaa aaacaccacg gaacctcatc tgttctcgta 1381 ctttgttgtg acaaagtagc tcactgcctt attatcacat tttcattatg caacgcttcg 1441 gaaaatacga tgttgaaaat ggatccggtt ccgcgcacat ttccccgaaa agtgccacct 1501 gggtcctttt catcacgtgc tataaaaata attataattt aaatttttta atataaatat 1561 ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt tgttttccga 1621 agatgtaaaa gactctaggg ggatcgccaa caaatactac cttttatctt gctcttcctg 1681 ctctcaggta ttaatgccga attgtttcat cttgtctgtg tagaagacca cacacgaaaa 1741 tcctgtgatt ttacatttta cttatcgtta atcgaatgta tatctattta atctgctttt 1801 cttgtctaat aaatatatat gtaaagtacg ctttttgttg aaatttttta aacctttgtt 1861 tatttttttt tcttcattcc gtaactcttc taccttcttt atttactttc taaaatccaa 1921 atacaaaaca taaaaataaa taaacacaga gtaaattccc aaattattcc atcattaaaa 1981 gatacgaggc gcgtgtaagt tacaggcaag cgatccgggt accctcgcat gcattaaggg 2041 ttctcgagag ctcgttttcg acactggatg gcggcgttag tatcgaatcg acagcagtat 2101 agcgaccagc attcacatac gattgacgca tgatattact ttctgcgcac ttaacttcgc 2161 atctgggcag atgatgtcga ggcgaaaaaa aatataaatc acgctaacat ttgattaaaa 2221 tagaacaact acaatataaa aaaactatac aaatgacaag ttcttgaaaa caagaatctt 2281 tttattgtca gtactgatta gaaaaactca tcgagcatca aatgaaactg caatttattc 2341 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac 2401 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt 2461 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa 2521 tcaccatgag tgacgactga atccggtgag aatggcaaaa gcttatgcat ttctttccag 2581 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg 2641 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa 2701 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt 2761 tcacctgaat caggatattc ttctaatacc tggaatgctg ttttgccggg gatcgcagtg 2821 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata 2881 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct 2941 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc 3001 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg 3061 ttggaattta atcgcggcct cgaaacgtga gtcttttcct tacccatggt tgtttatgtt 3121 cggatgtgat gtgagaactg tatcctagca agattttaaa aggaagtata tgaaagaaga 3181 acctcagtgg caaatcctaa ccttttatat ttctctacag gggcgcggcg tggggacaat 3241 tcaacgcgtc tgtgagggga gcgtttccct gctcgcaggt ctgcagcgag gagccgtaat 3301 ttttgcttcg cgccgtgcgg ccatcaaaat gtatggatgc aaatgattat acatggggat 3361 gtatgggcta aatgtacggg cgacagtcac atcatgcccc tgagctgcgc acgtcaagac 3421 tgtcaaggag ggtattctgg gcctccatgt cgctggccgg gtgacccggc ggggacgagg 3481 caagctaaac agatctctag acctatctaa tctagcgcga cgtctttaat taagcggccg 3541 ccatcactga ccatttaaat catacctgac ctccatagca gaaagtcaaa agcctccgac 3601 cggaggcttt tgacttgatc ggcacgtaag aggttccaac tttcaccata atgaaataag 3661 atcactaccg ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat 3721 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 3781 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 3841 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt tacgccccga 3901 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 3961 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 4021 tgagtactca ccagtcacag aaaagcatct cacggatggc atgacagtaa gagaattatg 4081 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctgg caacgatcgg 4141 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 4201 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 4261 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 4321 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggatcac ttctgcgctc 4381 ggccctcccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 4441 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgcatcgtag ttatctacac 4501 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 4561 actgattaag cattggtaag tgaccaaaca ggaaaaaacc gcccttaaca tggcccgctt 4621 tatcagaagc cagacattaa cgcttctgga gaaactcaac gagctggacg cggatgaaca 4681 ggcagacatc tgtgaatcgc ttcacgacca cgctgatgag ctttaccgca gctgcctcgc 4741 gcgtttcggt gatgacggtg aaaacctctg atgagggccc aaatgtaatc acctggctca 4801 ccttcgggtg ggcctttctg cgttgctggc gtttttccat aggctccgcc cccctgacga 4861 gcatcacaaa aatcgatgct caagtcagag gtggcgaaac ccgacaggac tataaagata 4921 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 4981 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 5041 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 5101 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca accaggtaag 5161 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 5221 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 5281 atttggtatc tgcgctctgc tgaagccagt tacctcggaa aaagagttgg tagctcttga 5341 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 5401 cgcagaaaaa aaggatctca agaagatcct ttgattttct accgaagaaa ggcccacccg 5461 tgaaggtgag ccagtgagtt gattgcagtc cagttacgct ggagtcaagc gcggccgcgg 5521 cggtctcggc attcctgctg aa //