LOCUS       sequence-274474-        6049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  high-copy URA-selectable plasmid, WT ECM33 intron (stable) inserted 
            into URA3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6049)
  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 6049)
  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-ECM_stable
FEATURES             Location/Qualifiers
     source          1..6049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     88..107
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        131..346
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     rep_origin      complement(1611..2066)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1748..1769)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1960..1979
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2192..2214
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2206..2223
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2207..2223
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2233..2251
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2253..2273)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2253..2271)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2292..2308)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2292..2308)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2305..2327)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2316..2332
                     /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(2340..2346,2347..2364,2365..2370))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2385..2406
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2523..2540)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2694..3282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2774..2793)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3453..4244,4245..4313))
                     /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     4076..4095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4314..4418)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5038..5915
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     5952..5970
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6012..6034)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
       61 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtaccg agagtgcacc
      121 ataccacctt ttcaattcat catttttttt ttattctttt ttttgatttc ggtttccttg
      181 aaattttttt gattcggtaa tctccgaaca gaaggaagaa cgaaggaagg agcacagact
      241 tagattggta tatatacgca tatgtagtgt tgaagaaaca tgaaattgcc cagtattctt
      301 aacccaactg cacagaacaa aaacctgcag gaaacgaaga taaatcatgt cgaaagctac
      361 atataaggaa cgtgctgcta ctcatcctag tcctgttggt atgtacacat tctccttttt
      421 tttcatcttt ttttctttat ttgcctcttt cctcacaaat ctcgagtaga ttcgtggtcc
      481 tcttcattct tttctttttt ctttgtcgat tactgggctt tttttcatag gtctcgattg
      541 acgcggacgg acaatgcgaa aaaaaaaaat ttccaaaaga ggaaacgggt ttcgagatct
      601 cgtgaggatg gttttggaat attcttcaat atttctgcct gtcataggat tagggcgagt
      661 aatgaaacag aataatacat gtataaatcg atcgggaata ctaacactac ttttctttat
      721 ctaagcagct gccaagctat ttaatatcat gcacgaaaag caaacaaact tgtgtgcttc
      781 attggatgtt cgtaccacca aggaattact ggagttagtt gaagcattag gtcccaaaat
      841 ttgtttacta aaaacacatg tggatatctt gactgatttt tccatggagg gcacagttaa
      901 gccgctaaag gcattatccg ccaagtacaa ttttttactc ttcgaagaca gaaaatttgc
      961 tgacattggt aatacagtca aattgcagta ctctgcgggt gtatacagaa tagcagaatg
     1021 ggcagacatt acgaatgcac acggtgtggt gggcccaggt attgttagcg gtttgaagca
     1081 ggcggcagaa gaagtaacaa aggaacctag aggccttttg atgttagcag aattgtcatg
     1141 caagggctcc ctatctactg gagaatatac taagggtact gttgacattg cgaagagcga
     1201 caaagatttt gttatcggct ttattgctca aagagacatg ggtggaagag atgaaggtta
     1261 cgattggttg attatgacac ccggtgtggg tttagatgac aagggtgatg cattgggtca
     1321 acagtataga accgtggatg atgtggtctc tacaggatct gacattatta ttgttggaag
     1381 aggactattt gcaaagggaa gggatgctaa ggtagagggt gaacgttaca gaaaagcagg
     1441 ctgggaagca tatttgagaa gatgcggcca gcaaaactaa aaaactgtat tataagtaaa
     1501 tgcatgtata ctaaactcac aaattagagc ttcaatttaa ttatatcagt tattacccta
     1561 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag
     1621 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     1681 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     1741 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     1801 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     1861 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     1921 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     1981 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     2041 gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa
     2101 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     2161 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     2221 agtgagcgcg cgtaatacga ctcactatag gccctttagt gagggttaat tgcgcgcttg
     2281 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     2341 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     2401 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     2461 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     2521 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2581 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2641 acaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2701 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2761 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2821 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2881 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2941 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3001 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3061 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3121 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3181 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3241 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3301 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3361 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3421 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3481 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3541 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3601 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3661 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3721 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     3781 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3841 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3901 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3961 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4021 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4081 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4141 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4201 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4261 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4321 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4381 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     4441 cctgcttatc gatgataagc tgtcaaacat gagaattaat tccacggact atagactata
     4501 cctagtatac tccgtctact gtacgataca cttccgctca ggtccttgtc ctttaacgag
     4561 gccttaccac tcttttgtta ctctattgat ccagctcagc aaaggcagtg tgatctaaga
     4621 ttctatcttc gcgatgtagt aaaactagct agaccgagaa agagactaga aatgcaaaag
     4681 gcacttctac aatggctgcc atcattatta tccgatgtga cgctgcagct tctcaatgat
     4741 attcgaatac gctttgagga gatacagcct aatatccgac aaactgtttt acagatttac
     4801 gatcgtactt gttacccatc attgaatttt gaacatccga acctgggagt tttccctgaa
     4861 acagatagta tatttgaacc tgtataataa tatatagtct agcgctttac ggaagacaat
     4921 gtatgtattt cggttcctgg agaaactatt gcatctattg cataggtaat cttgcacgtc
     4981 gcatccccgg ttcattttct gcgtttccat cttgcacttc aatagcatat ctttgttaac
     5041 gaagcatctg tgcttcattt tgtagaacaa aaatgcaacg cgagagcgct aatttttcaa
     5101 acaaagaatc tgagctgcat ttttacagaa cagaaatgca acgcgaaagc gctattttac
     5161 caacgaagaa tctgtgcttc atttttgtaa aacaaaaatg caacgcgaga gcgctaattt
     5221 ttcaaacaaa gaatctgagc tgcattttta cagaacagaa atgcaacgcg agagcgctat
     5281 tttaccaaca aagaatctat acttcttttt tgttctacaa aaatgcatcc cgagagcgct
     5341 atttttctaa caaagcatct tagattactt tttttctcct ttgtgcgctc tataatgcag
     5401 tctcttgata actttttgca ctgtaggtcc gttaaggtta gaagaaggct actttggtgt
     5461 ctattttctc ttccataaaa aaagcctgac tccacttccc gcgtttactg attactagcg
     5521 aagctgcggg tgcatttttt cgagataaag gcatccccga ttatattcta taccgatgtg
     5581 gattgcgcat actttgtgaa cagaaagtga tagcgttgat gattcttcat tggtcagaaa
     5641 attatgaacg gtttcttcta ttttgtctct atatactacg tataggaaat gtttacattt
     5701 tcgtattgtt ttcgattcac tctatgaata gttcttacta caattttttt gtctaaagag
     5761 taatactaga gataaacata aaaaatgtag aggtcgagtt tagatgcaag ttcaaggagc
     5821 gaaaggtgga tgggtaggtt atatagggat atagcacaga gatatatagc aaagagatac
     5881 ttttgagcaa tgtttgtgga agcggtattc gcaattaaga aaccattatt atcatgacat
     5941 taacctataa aaataggcgt atcacgaggc cctttcgtcg aattcgcgcg tttcggtgat
     6001 gacggtgaaa acctctgaca catgcagctc ccggagatct cggtcacag
//
