LOCUS       sequence-332939-        6530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  WT Ecm2 mutated to introduce stop codon after AA 325 of Ecm2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6530)
  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 6530)
  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 1-325
FEATURES             Location/Qualifiers
     source          1..6530
                     /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..2847)
                     /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(3018..3809,3810..3878))
                     /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     3641..3660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3879..3983)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4020..4523
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4564..4582
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4620..4642)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4742..4761
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4785..5000
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             5001..5804
                     /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(5935..6390)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6072..6093)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6284..6303
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(6516..6530,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6530,1..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 aatatacttc tgcgagcaac atcagatggc ccaagtactc
      841 cgttaataaa aaacacaact gcattgtata aaaatgaaaa aggtgcaaat gaggtcaaaa
      901 acttggaaaa gtacgcgtcc gtggacattt ctcatatttt gaagaaacta cccttgaatg
      961 aatcgttttt aaaaaacccc tccaccaaat cattttttct gtataatatt gatgcctcga
     1021 tccctgaatg gaaaataact gatacagttt cgcaattatt gggtataaag aaatggaagg
     1081 atggaaattc attgtcatta atagtcaacc acaaggcgaa gtgtggtggc ctaagatttc
     1141 aatctagtga attgggggag cggtttgtca gtaaaataag tgagacactt gtcacaccaa
     1201 agggcctaaa gagaggagtt ttacttattg accgttttag aatctttatc attccgtggt
     1261 cgtctggctt ttcagctgcg tcattcggaa ccaacactgc agaaaatata aaacttagtt
     1321 taagtttgaa taagcttatt cagttggagc tagggtaata gtaatctttt 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 tttttgtttg caagcagcag
     2821 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2881 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     2941 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3001 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3061 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3121 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3181 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3241 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3301 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3361 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3421 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3481 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3541 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3601 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3661 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     3721 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     3781 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     3841 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     3901 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     3961 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctgg gtccttttca
     4021 tcacgtgcta taaaaataat tataatttaa attttttaat ataaatatat aaattaaaaa
     4081 tagaaagtaa aaaaagaaat taaagaaaaa atagtttttg ttttccgaag atgtaaaaga
     4141 ctctaggggg atcgccaaca aatactacct tttatcttgc tcttcctgct ctcaggtatt
     4201 aatgccgaat tgtttcatct tgtctgtgta gaagaccaca cacgaaaatc ctgtgatttt
     4261 acattttact tatcgttaat cgaatgtata tctatttaat ctgcttttct tgtctaataa
     4321 atatatatgt aaagtacgct ttttgttgaa attttttaaa cctttgttta tttttttttc
     4381 ttcattccgt aactcttcta ccttctttat ttactttcta aaatccaaat acaaaacata
     4441 aaaataaata aacacagagt aaattcccaa attattccat cattaaaaga tacgaggcgc
     4501 gtgtaagtta caggcaagcg atccgtctaa gaaaccatta ttatcatgac attaacctat
     4561 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     4621 ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc
     4681 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat
     4741 gcggcatcag agcagattgt actgagagtg caccatacca ccttttcaat tcatcatttt
     4801 ttttttattc ttttttttga tttcggtttc cttgaaattt ttttgattcg gtaatctccg
     4861 aacagaagga agaacgaagg aaggagcaca gacttagatt ggtatatata cgcatatgta
     4921 gtgttgaaga aacatgaaat tgcccagtat tcttaaccca actgcacaga acaaaaacct
     4981 gcaggaaacg aagataaatc atgtcgaaag ctacatataa ggaacgtgct gctactcatc
     5041 ctagtcctgt tgctgccaag ctatttaata tcatgcacga aaagcaaaca aacttgtgtg
     5101 cttcattgga tgttcgtacc accaaggaat tactggagtt agttgaagca ttaggtccca
     5161 aaatttgttt actaaaaaca catgtggata tcttgactga tttttccatg gagggcacag
     5221 ttaagccgct aaaggcatta tccgccaagt acaatttttt actcttcgaa gacagaaaat
     5281 ttgctgacat tggtaataca gtcaaattgc agtactctgc gggtgtatac agaatagcag
     5341 aatgggcaga cattacgaat gcacacggtg tggtgggccc aggtattgtt agcggtttga
     5401 agcaggcggc agaagaagta acaaaggaac ctagaggcct tttgatgtta gcagaattgt
     5461 catgcaaggg ctccctatct actggagaat atactaaggg tactgttgac attgcgaaga
     5521 gcgacaaaga ttttgttatc ggctttattg ctcaaagaga catgggtgga agagatgaag
     5581 gttacgattg gttgattatg acacccggtg tgggtttaga tgacaaggga gacgcattgg
     5641 gtcaacagta tagaaccgtg gatgatgtgg tctctacagg atctgacatt attattgttg
     5701 gaagaggact atttgcaaag ggaagggatg ctaaggtaga gggtgaacgt tacagaaaag
     5761 caggctggga agcatatttg agaagatgcg gccagcaaaa ctaaaaaact gtattataag
     5821 taaatgcatg tatactaaac tcacaaatta gagcttcaat ttaattatat cagttattac
     5881 cctatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg
     5941 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     6001 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     6061 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     6121 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     6181 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     6241 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     6301 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     6361 ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg
     6421 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     6481 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
//
