LOCUS       sequence-332936-        6529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  WT Ecm2 mutated to introduce stop codon after AA 143 of Ecm2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6529)
  AUTHORS   van der Feltz C, Nikolai B, Schneider C, Paulson JC, Fu X, Hoskins 
            AA
  TITLE     Saccharomyces cerevisiae Ecm2 Modulates the Catalytic Steps of 
            pre-mRNA Splicing.
  JOURNAL   RNA. 2021 Feb 5. pii: rna.077727.120. doi: 10.1261/rna.077727.120.
  PUBMED    33547186
REFERENCE   2  (bases 1 to 6529)
  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"; journalName: "RNA. 2021 
            Feb 5. pii: rna.077727.120. doi: 10.1261/rna.077727.120."
COMMENT     Sequence Label: Ecm2 1-143
FEATURES             Location/Qualifiers
     source          1..6529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1818..1838)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1818..1836)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1857..1873)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1857..1873)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1870..1892)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1881..1897
                     /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(1905..1911,1912..1929,1930..1935))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1950..1971
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2088..2105)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2259..2846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2339..2358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3017..3808,3809..3877))
                     /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     3640..3659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3878..3982)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4019..4522
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4563..4581
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4619..4641)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4741..4760
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4784..4999
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5000..5803
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(5934..6389)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6071..6092)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6283..6302
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(6515..6529,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6529,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg
       61 ggccccccct cgaggtcgac gataaatagt aataaaacta agcgggttag taatactcac
      121 caagaacaat agcaggaacg atagtcggtt ctctttcgta atttttttaa gtcacaaccc
      181 ggagggacgt gaagtttcaa accttattca ttccgggatt accctctcct gtgatgacgt
      241 ttaggcaata aggactaaaa aaaaaaaaaa gaaatgctat ggcaaatttt attgatagaa
      301 gtggtaatga tgatggaagg gcttttgagc aggtgaatag aaggtagcta acggaaaaga
      361 gttattgcac ccttaatata atgaatgatg aaattaatga gccaccgccc aatatatgtg
      421 agcagtgctt aggtgatgag gctaatatac gaatgactaa aattccacaa ggctctgaat
      481 gtaagatttg taccttaccg ttcaccttat accattttaa gacgtctaaa cgaagcaata
      541 atatcattaa aacattaata tgtgtacggt gcgccactca gagaaacatt tgtcaatgct
      601 gtatgcttga ttcaagatgg cacatcccta tacaactgcg ggatcattta atatcccttg
      661 tgaacgagga aaacgttatg acggaggaag caaaaaacga catgatgaaa aggtttctgt
      721 cactaaaaaa cgtgaaatta ggaggagctc agattacgag tgatccgtca gaagcagata
      781 atatcgttga taaattgaaa aatatacttt aatagtaacg agcaacatca gatggcccaa
      841 gtactccgtt aataaaaaac acaactgcat tgtataaaaa tgaaaaaggt gcaaatgagg
      901 tcaaaaactt ggaaaagtac gcgtccgtgg acatttctca tattttgaag aaactaccct
      961 tgaatgaatc gtttttaaaa aacccctcca ccaaatcatt ttttctgtat aatattgatg
     1021 cctcgatccc tgaatggaaa ataactgata cagtttcgca attattgggt ataaagaaat
     1081 ggaaggatgg aaattcattg tcattaatag tcaaccacaa ggcgaagtgt ggtggcctaa
     1141 gatttcaatc tagtgaattg ggggagcggt ttgtcagtaa aataagtgag acacttgtca
     1201 caccaaaggg cctaaagaga ggagttttac ttattgaccg ttttagaatc tttatcattc
     1261 cgtggtcgtc tggcttttca gctgcgtcat tcggaaccaa cactgcagaa aatataaaac
     1321 ttagtttaag tttgaataag cttattcagt tggagctagg gctttctttt cctacaaaaa
     1381 gcacagataa cgcaaagaat gacaagaaaa agacatcaaa aaaagttcat aaggacagat
     1441 caaagaaatc gaaacctcgc gctaacaagt taacaatata gatgaaacgt aagaaaaatc
     1501 atgggttgaa gaaacgaact gtttgatcaa tttcgaccat ccatctacct tgcaagtaat
     1561 aaacaaaagc aggttgagtg gggaacttta tctaagttga tcagtatatc tgacaaattt
     1621 atacggaatg ctgcagtttg gattagcacg aacattttat ctgtacggtt tgtcgctttt
     1681 agctgctcgg gctcttcctc tcggttaccg agatgataac tcggccgaat tttgtaagta
     1741 caaataaaaa gaaagtgaca caattctaaa agtatcacta tgcggccgcc accgcggtgg
     1801 agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     1861 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1921 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     1981 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2041 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2101 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2161 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2221 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2281 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2341 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2401 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2461 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2521 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2581 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2641 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2701 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2761 tcttgatccg gcaaacaaac caccgctggt agcggtggtt ttttgtttgc aagcagcaga
     2821 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2881 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2941 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3001 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3061 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3121 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3181 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3241 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3301 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3361 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3421 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3481 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3541 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3601 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3661 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3721 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3781 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3841 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     3901 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     3961 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg tccttttcat
     4021 cacgtgctat aaaaataatt ataatttaaa ttttttaata taaatatata aattaaaaat
     4081 agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt tttccgaaga tgtaaaagac
     4141 tctaggggga tcgccaacaa atactacctt ttatcttgct cttcctgctc tcaggtatta
     4201 atgccgaatt gtttcatctt gtctgtgtag aagaccacac acgaaaatcc tgtgatttta
     4261 cattttactt atcgttaatc gaatgtatat ctatttaatc tgcttttctt gtctaataaa
     4321 tatatatgta aagtacgctt tttgttgaaa ttttttaaac ctttgtttat ttttttttct
     4381 tcattccgta actcttctac cttctttatt tactttctaa aatccaaata caaaacataa
     4441 aaataaataa acacagagta aattcccaaa ttattccatc attaaaagat acgaggcgcg
     4501 tgtaagttac aggcaagcga tccgtctaag aaaccattat tatcatgaca ttaacctata
     4561 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     4621 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     4681 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     4741 cggcatcaga gcagattgta ctgagagtgc accataccac cttttcaatt catcattttt
     4801 tttttattct tttttttgat ttcggtttcc ttgaaatttt tttgattcgg taatctccga
     4861 acagaaggaa gaacgaagga aggagcacag acttagattg gtatatatac gcatatgtag
     4921 tgttgaagaa acatgaaatt gcccagtatt cttaacccaa ctgcacagaa caaaaacctg
     4981 caggaaacga agataaatca tgtcgaaagc tacatataag gaacgtgctg ctactcatcc
     5041 tagtcctgtt gctgccaagc tatttaatat catgcacgaa aagcaaacaa acttgtgtgc
     5101 ttcattggat gttcgtacca ccaaggaatt actggagtta gttgaagcat taggtcccaa
     5161 aatttgttta ctaaaaacac atgtggatat cttgactgat ttttccatgg agggcacagt
     5221 taagccgcta aaggcattat ccgccaagta caatttttta ctcttcgaag acagaaaatt
     5281 tgctgacatt ggtaatacag tcaaattgca gtactctgcg ggtgtataca gaatagcaga
     5341 atgggcagac attacgaatg cacacggtgt ggtgggccca ggtattgtta gcggtttgaa
     5401 gcaggcggca gaagaagtaa caaaggaacc tagaggcctt ttgatgttag cagaattgtc
     5461 atgcaagggc tccctatcta ctggagaata tactaagggt actgttgaca ttgcgaagag
     5521 cgacaaagat tttgttatcg gctttattgc tcaaagagac atgggtggaa gagatgaagg
     5581 ttacgattgg ttgattatga cacccggtgt gggtttagat gacaagggag acgcattggg
     5641 tcaacagtat agaaccgtgg atgatgtggt ctctacagga tctgacatta ttattgttgg
     5701 aagaggacta tttgcaaagg gaagggatgc taaggtagag ggtgaacgtt acagaaaagc
     5761 aggctgggaa gcatatttga gaagatgcgg ccagcaaaac taaaaaactg tattataagt
     5821 aaatgcatgt atactaaact cacaaattag agcttcaatt taattatatc agttattacc
     5881 ctatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     5941 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     6001 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     6061 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     6121 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6181 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6241 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6301 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     6361 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     6421 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     6481 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgtt
//
