LOCUS       sequence-332935-        6524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ecm2 +/-300bp of up- and downstream seq. cloned into the NotI and 
            SalI sites of pRS416.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6524)
  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 6524)
  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: WT Ecm2
FEATURES             Location/Qualifiers
     source          1..6524
                     /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(1812..1832)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1812..1830)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1851..1867)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1851..1867)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1864..1886)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1875..1891
                     /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(1899..1905,1906..1923,1924..1929))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1944..1965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2082..2099)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2253..2841)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2333..2352)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3012..3803,3804..3872))
                     /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     3635..3654
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3873..3977)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    4014..4517
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     4558..4576
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4614..4636)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4736..4755
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        4779..4994
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4995..5798
                     /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(5929..6384)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6066..6087)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6278..6297
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(6510..6524,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6524,1..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 tagggctttc ttttcctaca aaaagcacag
     1381 ataacgcaaa gaatgacaag aaaaagacat caaaaaaagt tcataaggac agatcaaaga
     1441 aatcgaaacc tcgcgctaac aagttaacaa tatagatgaa acgtaagaaa aatcatgggt
     1501 tgaagaaacg aactgtttga tcaatttcga ccatccatct accttgcaag taataaacaa
     1561 aagcaggttg agtggggaac tttatctaag ttgatcagta tatctgacaa atttatacgg
     1621 aatgctgcag tttggattag cacgaacatt ttatctgtac ggtttgtcgc ttttagctgc
     1681 tcgggctctt cctctcggtt accgagatga taactcggcc gaattttgta agtacaaata
     1741 aaaagaaagt gacacaattc taaaagtatc actatgcggc cgccaccgcg gtggagctcc
     1801 agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg
     1861 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1921 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1981 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     2041 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2101 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2161 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2221 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2281 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2341 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2401 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2461 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2521 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2581 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2641 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2701 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2761 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2821 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2881 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2941 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     3001 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3061 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3121 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3181 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3241 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3301 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3361 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3421 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3481 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3541 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3601 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3661 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3721 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3781 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3841 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3901 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3961 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcctt ttcatcacgt
     4021 gctataaaaa taattataat ttaaattttt taatataaat atataaatta aaaatagaaa
     4081 gtaaaaaaag aaattaaaga aaaaatagtt tttgttttcc gaagatgtaa aagactctag
     4141 ggggatcgcc aacaaatact accttttatc ttgctcttcc tgctctcagg tattaatgcc
     4201 gaattgtttc atcttgtctg tgtagaagac cacacacgaa aatcctgtga ttttacattt
     4261 tacttatcgt taatcgaatg tatatctatt taatctgctt ttcttgtcta ataaatatat
     4321 atgtaaagta cgctttttgt tgaaattttt taaacctttg tttatttttt tttcttcatt
     4381 ccgtaactct tctaccttct ttatttactt tctaaaatcc aaatacaaaa cataaaaata
     4441 aataaacaca gagtaaattc ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa
     4501 gttacaggca agcgatccgt ctaagaaacc attattatca tgacattaac ctataaaaat
     4561 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     4621 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     4681 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca
     4741 tcagagcaga ttgtactgag agtgcaccat accacctttt caattcatca tttttttttt
     4801 attctttttt ttgatttcgg tttccttgaa atttttttga ttcggtaatc tccgaacaga
     4861 aggaagaacg aaggaaggag cacagactta gattggtata tatacgcata tgtagtgttg
     4921 aagaaacatg aaattgccca gtattcttaa cccaactgca cagaacaaaa acctgcagga
     4981 aacgaagata aatcatgtcg aaagctacat ataaggaacg tgctgctact catcctagtc
     5041 ctgttgctgc caagctattt aatatcatgc acgaaaagca aacaaacttg tgtgcttcat
     5101 tggatgttcg taccaccaag gaattactgg agttagttga agcattaggt cccaaaattt
     5161 gtttactaaa aacacatgtg gatatcttga ctgatttttc catggagggc acagttaagc
     5221 cgctaaaggc attatccgcc aagtacaatt ttttactctt cgaagacaga aaatttgctg
     5281 acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt atacagaata gcagaatggg
     5341 cagacattac gaatgcacac ggtgtggtgg gcccaggtat tgttagcggt ttgaagcagg
     5401 cggcagaaga agtaacaaag gaacctagag gccttttgat gttagcagaa ttgtcatgca
     5461 agggctccct atctactgga gaatatacta agggtactgt tgacattgcg aagagcgaca
     5521 aagattttgt tatcggcttt attgctcaaa gagacatggg tggaagagat gaaggttacg
     5581 attggttgat tatgacaccc ggtgtgggtt tagatgacaa gggagacgca ttgggtcaac
     5641 agtatagaac cgtggatgat gtggtctcta caggatctga cattattatt gttggaagag
     5701 gactatttgc aaagggaagg gatgctaagg tagagggtga acgttacaga aaagcaggct
     5761 gggaagcata tttgagaaga tgcggccagc aaaactaaaa aactgtatta taagtaaatg
     5821 catgtatact aaactcacaa attagagctt caatttaatt atatcagtta ttaccctatg
     5881 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg
     5941 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     6001 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     6061 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     6121 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     6181 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     6241 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     6301 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     6361 ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg
     6421 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6481 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgtt
//
