LOCUS       sequence-274482-        6008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  high-copy URA-selectable plasmid, Mut ACT1 intron (stable) inserted 
            into URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41586-018-0828-1"; journalName: "Nature"; date: "2019-01";
            volume: "565"; issue: "7741"; pages: "606-611"
COMMENT     Sequence Label: pRS426-ACT_stable
FEATURES             Location/Qualifiers
     source          1..6008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     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(join(1735..1741,1742..1759,1760..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(join(2848..3639,3640..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
//
