LOCUS Exported 5121 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION low-copy URA-selectable plasmid, ECM33 intron inserted into URA3. ACCESSION . VERSION . KEYWORDS pRS416-ECM33_intron SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5121) AUTHORS Morgan JT, Fink GR, Bartel DP TITLE Excised linear introns regulate growth in yeast. JOURNAL Nature. 2019 Jan;565(7741):606-611. doi: 10.1038/s41586-018-0828-1. Epub 2019 Jan 16. PUBMED 30651636 REFERENCE 2 (bases 1 to 5121) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5121 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 415..433 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(475..497) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 600..619 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 643..858 /gene="S. cerevisiae URA3" /label=URA3 promoter rep_origin complement(2123..2578) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2260..2281) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2472..2491 /label=F1ori-R /note="F1 origin, reverse primer" CDS complement(2555..2833) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSAQLTLTKGNKSWVPNSPYSESYYARSLAVVLQRRDWEN PGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWTRPVAAH" primer_bind 2704..2726 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 2718..2735 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 2719..2735 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 2745..2764 /label=T7 /note="T7 promoter, forward primer" promoter 2745..2763 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2790..2810) /label=T3 /note="T3 promoter, forward primer" promoter complement(2790..2808) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2829..2845) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2829..2845) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2842..2864) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2853..2869 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(2877..2907) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2922..2943 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3060..3077) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3231..3819) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3311..3330) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3990..4850) /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 4613..4632 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4851..4955) /gene="bla" /label=AmpR promoter misc_feature 4992..374 /label=CEN/ARS /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" ORIGIN 1 gatcgccaac aaatactacc ttttatcttg ctcttcctgc tctcaggtat taatgccgaa 61 ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat cctgtgattt tacattttac 121 ttatcgttaa tcgaatgtat atctatttaa tctgcttttc ttgtctaata aatatatatg 181 taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt cttcattccg 241 taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat aaaaataaat 301 aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg cgtgtaagtt 361 acaggcaagc gatccgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 421 cgtatcacga ggccctttcg tcgaattcgc gcgtttcggt gatgacggtg aaaacctctg 481 acacatgcag ctcccggaga tctcggtcac agcttgtctg taagcggatg ccgggagcag 541 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc 601 ggcatcagag cagattgtac cgagagtgca ccataccacc ttttcaattc atcatttttt 661 ttttattctt ttttttgatt tcggtttcct tgaaattttt ttgattcggt aatctccgaa 721 cagaaggaag aacgaaggaa ggagcacaga cttagattgg tatatatacg catatgtagt 781 gttgaagaaa catgaaattg cccagtattc ttaacccaac tgcacagaac aaaaacctgc 841 aggaaacgaa gataaatcat gtcgaaagct acatataagg aacgtgctgc tactcatcct 901 agtcctgttg gtatgtacac attctccttt tttttcatct ttttttcttt atttgcctct 961 ttcctcacaa atctcgagta gattcgtggt cctcttcatt cttttctttt ttctttgtcg 1021 attactgggc tttttttcat aggtctcgat tgacgcggac ggacaatgcg aaaaaaaaaa 1081 atttccaaaa gaggaaacgg gtttcgagat ctcgtgagga tggttttgga atattcttca 1141 atatttctgc ctgtcatagg attagggcga gtaatgaaac agaataatac atgtataaat 1201 cgatcgggaa tactaacact acttttcttt atctaagcag ctgccaagct atttaatatc 1261 atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac caaggaatta 1321 ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca tgtggatatc 1381 ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc cgccaagtac 1441 aattttttac tcttcgaaga cagaaaattt gctgacattg gtaatacagt caaattgcag 1501 tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc acacggtgtg 1561 gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac aaaggaacct 1621 agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac tggagaatat 1681 actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg ctttattgct 1741 caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac acccggtgtg 1801 ggtttagatg acaagggtga tgcattgggt caacagtata gaaccgtgga tgatgtggtc 1861 tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg aagggatgct 1921 aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag aagatgcggc 1981 cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc acaaattaga 2041 gcttcaattt aattatatca gttattaccc tatgcggtgt gaaataccgc acagatgcgt 2101 aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa attcgcgtta 2161 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat 2221 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca 2281 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc 2341 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta 2401 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg 2461 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg 2521 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc 2581 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat 2641 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt 2701 tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat 2761 agggcgaatt gggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg 2821 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac 2881 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca 2941 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat 3001 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc 3061 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 3121 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 3181 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 3241 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 3301 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 3361 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 3421 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 3481 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 3541 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 3601 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 3661 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 3721 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 3781 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 3841 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 3901 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 3961 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 4021 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 4081 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 4141 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 4201 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 4261 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg 4321 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 4381 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 4441 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 4501 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 4561 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc 4621 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 4681 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 4741 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 4801 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 4861 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 4921 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 4981 gggtcctttt catcacgtgc tataaaaata attataattt aaatttttta atataaatat 5041 ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt tgttttccga 5101 agatgtaaaa gactctaggg g //