LOCUS Exported 6008 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION high-copy URA-selectable plasmid, Mut ACT1 intron (stable) inserted into URA3. ACCESSION . VERSION . KEYWORDS pRS426-ACT_stable SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6008) 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 6008) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6008 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(1007..1462) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1144..1165) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1356..1375 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1588..1610 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1602..1619 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1603..1619 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 1629..1646 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1648..1668) /label=T3 /note="T3 promoter, forward primer" promoter complement(1648..1666) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1687..1703) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1687..1703) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1700..1722) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 1711..1727 /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(1735..1765) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 1780..1801 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1918..1935) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2089..2677) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2169..2188) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2848..3708) /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 3471..3490 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3709..3813) /gene="bla" /label=AmpR promoter rep_origin 4433..5310 /label=2u ori /note="yeast 2u plasmid origin of replication" primer_bind 5347..5365 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(5407..5429) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 5532..5551 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 5575..5790 /gene="S. cerevisiae URA3" /label=URA3 promoter ORIGIN 1 cttgaaattt ggaaaaaaaa aaaaaactga aatattttcg tgataagtga tagtgatatt 61 cttcttttat ttgctactgt tactaagtct catgtactaa catgttgcta tattatatgt 121 ttagctgcca agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg 181 gatgttcgta ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt 241 ttactaaaaa cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg 301 ctaaaggcat tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac 361 attggtaata cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca 421 gacattacga atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg 481 gcagaagaag taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag 541 ggctccctat ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa 601 gattttgtta tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat 661 tggttgatta tgacacccgg tgtgggttta gatgacaagg gtgatgcatt gggtcaacag 721 tatagaaccg tggatgatgt ggtctctaca ggatctgaca ttattattgt tggaagagga 781 ctatttgcaa agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg 841 gaagcatatt tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca 901 tgtatactaa actcacaaat tagagcttca atttaattat atcagttatt accctatgcg 961 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt 1021 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag 1081 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt 1141 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga 1201 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg 1261 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct 1321 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc 1381 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt 1441 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc 1501 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc 1561 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg 1621 agcgcgcgta atacgactca ctatagccct ttagtgaggg ttaattgcgc gcttggcgta 1681 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat 1741 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt 1801 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 1861 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 1921 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 1981 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgaacaaa 2041 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 2101 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 2161 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 2221 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 2281 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 2341 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 2401 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 2461 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 2521 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 2581 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 2641 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 2701 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 2761 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 2821 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 2881 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 2941 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 3001 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 3061 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 3121 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 3181 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 3241 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 3301 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 3361 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 3421 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 3481 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 3541 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 3601 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 3661 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 3721 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 3781 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgc 3841 ttatcgatga taagctgtca aacatgagaa ttaattccac ggactataga ctatacctag 3901 tatactccgt ctactgtacg atacacttcc gctcaggtcc ttgtccttta acgaggcctt 3961 accactcttt tgttactcta ttgatccagc tcagcaaagg cagtgtgatc taagattcta 4021 tcttcgcgat gtagtaaaac tagctagacc gagaaagaga ctagaaatgc aaaaggcact 4081 tctacaatgg ctgccatcat tattatccga tgtgacgctg cagcttctca atgatattcg 4141 aatacgcttt gaggagatac agcctaatat ccgacaaact gttttacaga tttacgatcg 4201 tacttgttac ccatcattga attttgaaca tccgaacctg ggagttttcc ctgaaacaga 4261 tagtatattt gaacctgtat aataatatat agtctagcgc tttacggaag acaatgtatg 4321 tatttcggtt cctggagaaa ctattgcatc tattgcatag gtaatcttgc acgtcgcatc 4381 cccggttcat tttctgcgtt tccatcttgc acttcaatag catatctttg ttaacgaagc 4441 atctgtgctt cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa 4501 gaatctgagc tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg 4561 aagaatctgt gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa 4621 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac 4681 caacaaagaa tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt 4741 tctaacaaag catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct 4801 tgataacttt ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt 4861 ttctcttcca taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct 4921 gcgggtgcat tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg 4981 cgcatacttt gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat 5041 gaacggtttc ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta 5101 ttgttttcga ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata 5161 ctagagataa acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag 5221 gtggatgggt aggttatata gggatatagc acagagatat atagcaaaga gatacttttg 5281 agcaatgttt gtggaagcgg tattcgcaat taagaaacca ttattatcat gacattaacc 5341 tataaaaata ggcgtatcac gaggcccttt cgtcgaattc gcgcgtttcg gtgatgacgg 5401 tgaaaacctc tgacacatgc agctcccgga gatctcggtc acagcttgtc tgtaagcgga 5461 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg 5521 gcttaactat gcggcatcag agcagattgt accgagagtg caccatacca ccttttcaat 5581 tcatcatttt ttttttattc ttttttttga tttcggtttc cttgaaattt ttttgatttg 5641 gtaatctccg aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata 5701 cgcatatgta gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga 5761 acaaaaacct gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct 5821 gctactcatc ctagtcctgt tggtatgttc tagcgcttgc accatcccat ttaactgtaa 5881 gaagaattgc acggtcccaa ttgctcgaga gatttctctt ttaccttttt ttactatttt 5941 tcactctccc ataacctcct atattgactg atctgtaata accacgatat tattggaata 6001 aatagggg //