LOCUS       sequence-274481-        6027 bp    DNA     circular SYN 15-APR-2025
DEFINITION  high-copy URA-selectable plasmid, WT ACT1 intron (unstable) inserted
            into URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6027)
  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 6027)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_unstable
FEATURES             Location/Qualifiers
     source          1..6027
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        167..380
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      complement(1625..2080)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1762..1783)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1974..1993
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2206..2228
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2220..2237
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2221..2237
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2247..2265
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2267..2287)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2267..2285)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2306..2322)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2306..2322)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2319..2341)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2330..2346
                     /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(2354..2360,2361..2378,2379..2384))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2399..2420
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2537..2554)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2708..3296)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2788..2807)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3467..4258,4259..4327))
                     /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     4090..4109
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4328..4432)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5052..5929
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     5966..5984
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(6026..6027,1..21))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tctgacacat gcagctcccg gagatctcgg tcacagcttg tctgtaagcg gatgccggga
       61 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
      121 atgcggcatc agagcagatt gtaccgagag tgcaccatac caccttttca attcatcatt
      181 ttttttattc ttttttttga tttcggtttc cttgaaattt ttttgattcg gtaatctccg
      241 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta
      301 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacct
      361 gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc
      421 ctagtcctgt tggtatgttc tagcgcttgc accatcccat ttaactgtaa gaagaattgc
      481 acggtcccaa ttgctcgaga gatttctctt ttaccttttt ttactatttt tcactctccc
      541 ataacctcct atattgactg atctgtaata accacgatat tattggaata aataggggct
      601 tgaaatttgg aaaaaaaaaa aaaaactgaa atattttcgt gataagtgat agtgatattc
      661 ttcttttatt tgctactgtt actaagtctc atgtactaac atcgattgct tcattctttt
      721 tgttgctata ttatatgttt agctgccaag ctatttaata tcatgcacga aaagcaaaca
      781 aacttgtgtg cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca
      841 ttaggtccca aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg
      901 gagggcacag ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgaa
      961 gacagaaaat ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac
     1021 agaatagcag aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt
     1081 agcggtttga agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta
     1141 gcagaattgt catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac
     1201 attgcgaaga gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga
     1261 agagatgaag gttacgattg gttgattatg acacccggtg tgggtttaga tgacaagggt
     1321 gatgcattgg gtcaacagta tagaaccgtg gatgatgtgg tctctacagg atctgacatt
     1381 attattgttg gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt
     1441 tacagaaaag caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact
     1501 gtattataag taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat
     1561 cagttattac cctatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     1621 caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     1681 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     1741 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     1801 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     1861 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     1921 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     1981 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     2041 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc
     2101 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     2161 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     2221 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact ataggccctt tagtgagggt
     2281 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     2341 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     2401 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     2461 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     2521 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     2581 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     2641 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     2701 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2761 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2821 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2881 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2941 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3001 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3061 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3121 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3181 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3241 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3301 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3361 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3421 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     3481 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     3541 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     3601 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     3661 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     3721 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     3781 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     3841 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     3901 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     3961 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4021 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4081 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4141 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4201 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4261 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4321 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     4381 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     4441 cccgaaaagt gccacctgct tatcgatgat aagctgtaaa acatgagaat taattccacg
     4501 gactatagac tatacctagt atactccgtc tactgtacga tacacttccg ctcaggtcct
     4561 tgtcctttaa cgaggcctta ccactctttt gttactctat tgatccagct cagcaaaggc
     4621 agtgtgatct aagattctat cttcgcgatg tagtaaaact agctagaccg agaaagagac
     4681 tagaaatgca aaaggcactt ctacaatggc tgccatcatt attatccgat gtgacgctgc
     4741 agcttctcaa tgatattcga atacgctttg aggagataca gcctaatatc cgacaaactg
     4801 ttttacagat ttacgatcgt acttgttacc catcattgaa ttttgaacat ccgaacctgg
     4861 gagttttccc tgaaacagat agtatatttg aacctgtata ataatatata gtctagcgct
     4921 ttacggaaga caatgtatgt atttcggttc ctggagaaac tattgcatct attgcatagg
     4981 taatcttgca catcgcatcc ccggttcatt ttctgcgttt ccatcttgca cttcaatagc
     5041 atatctttgt taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag
     5101 cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga
     5161 aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc
     5221 gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa
     5281 cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc
     5341 atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc
     5401 gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa
     5461 ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt
     5521 actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat
     5581 tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct
     5641 tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg
     5701 aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt
     5761 ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg
     5821 caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata
     5881 tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaatt aagaaaccat
     5941 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcgaattcg
     6001 cgcgtttcgg tgatgacggt gaaaacc
//
