LOCUS       sequence-170836-        8524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Prp5 E235A for expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8524)
  AUTHORS   Carrocci TJ, Zoerner DM, Paulson JC, Hoskins AA
  TITLE     SF3b1 mutations associated with myelodysplastic syndromes alter the 
            fidelity of branchsite selection in yeast.
  JOURNAL   Nucleic Acids Res. 2017 Jan 6. pii: gkw1349. doi: 
            10.1093/nar/gkw1349.
  PUBMED    28062854
REFERENCE   2  (bases 1 to 8524)
  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: "Nucleic 
            Acids Res. 2017 Jan 6. pii: gkw1349. doi: 10.1093/nar/gkw1349."
COMMENT     Sequence Label: pRS413 Prp5 E235A
FEATURES             Location/Qualifiers
     source          1..8524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    70..573
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        610..714
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(715..783,784..1575)
                     /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     complement(933..952)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1746..2334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2235..2254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2488..2505
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(2658..2663,2664..2681,2682..2688)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2696..2712
                     /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)."
     primer_bind     2701..2723
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2720..2736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2720..2736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2755..2775
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2757..2775
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6416..6432)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6417..6433)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(6457..6476)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6458..6476)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6486..6503)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6486..6502)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6495..6517)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6647..7102
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6734..6753)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6944..6965
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7362..8021)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(8022..8208)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(8355..8374)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8474..8496
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata
      121 atttgggaat ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt
      181 aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa
      241 atttcaacaa aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata
      301 gatatacatt cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct
      361 tctacacaga caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata
      421 aaaggtagta tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact
      481 attttttctt taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa
      541 atttaaatta taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg
      601 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      661 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      721 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      781 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      841 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      901 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      961 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1021 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1081 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1141 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1201 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1261 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1321 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1381 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1441 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1501 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1561 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1621 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1681 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1741 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1801 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1861 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
     1921 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1981 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     2041 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     2101 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2161 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2221 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2281 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2341 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2401 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2461 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2521 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     2581 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact
     2641 cattaggcac cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg
     2701 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
     2761 aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg ctctagaatt
     2821 taaacccgga ttctgcctca atgggtagat cagacacctt agaagtaagg atagagctgc
     2881 tttgcaaaac tggtaatgat ccattaccat caccaggcgc attggcagta actttggagt
     2941 ctttttttga gactgtggac gattcatcac cagtaattga gccgtactga cggtcatagt
     3001 cttcttgtga catccatatt ggcttctctg gtttggttga cattctactt ttctaagacg
     3061 gataatggac gactccacgt tgtagtagtt gtcacattaa tcggaaaagt gtctcacatg
     3121 taaaggatta aaataagaaa tcttctaatt tatgttgaaa tatcttgatt taccattacc
     3181 cggattagat gaaactaatt aaagtcattt agagcaatgg accaaactaa aggtattaga
     3241 gttctttcta atataataca gcgccaaata attcgagaag ggaaatcata acgctgttgc
     3301 cttgggttta tttgaggtat ggaaactatt gattcgaagc aaaatattaa tagggagtct
     3361 ttattggagg aaaggaggaa aaaactagca aaatggaaac aaaaaaaagc acaatttgat
     3421 gctcagaaag agcatcaaac ttcacgcaat gacattgtta ctaacagttt agaaggtaag
     3481 cagactactg aaaagtttac ggaaagacag gaacgtgtaa aagaagagct tcggaagaga
     3541 aaaaatgaat ttaggaaatc tgacgaacct gtatcggtta agccttcgaa gaaaaagtcg
     3601 aagagaagta aggtaaagaa gaaaatatcc ttcgacttca gtgatgacga tgattctgaa
     3661 ataggagttt cttttagatc gaaggagcac attcaaaaag ctccagagca tgataatgaa
     3721 aaagatccac tggatgagtt tatgacgtca ttgaaggagg agaaaatgag taatagtaaa
     3781 ggcatgtacg acagaggcga cattcttgat gtagaggatc aattgtttga acttggagga
     3841 accgacgatg aggatgttga agataatacg gataattcta atatagcgaa aattgctaaa
     3901 ctcaaagcaa aaaaacgtgt aaagcaaatt tattactctc cagaagagct tgagccgttc
     3961 caaaaaaatt tttatataga atctgaaaca gtttcctcga tgtcagagat ggaagttgag
     4021 gcacttagac ttagtctgga taatatcaaa ataaagggaa caggttgccc caaaccagtc
     4081 acaaagtggt ctcaactggg actatcaacg gacaccatgg ttttaattac agaaaagttg
     4141 cactttggct ccttaacgcc tattcaatcc caggctcttc ccgctattat gtcaggtcgt
     4201 gatgttatag gaatatcaaa aactggctcc ggcaagacca tctcctatct tttaccattg
     4261 ctaagacaag taaaagctca acggccatta tcaaaacatg aaacagggcc catgggttta
     4321 attctggctc caactagaga gctagcttta caaatacatg aagaagttac taaattcaca
     4381 gaagcagata catccattag atcagtatgc tgtacaggag gttctgaaat gaaaaagcag
     4441 attactgatc ttaaaagagg cactgagatt gttgttgcca caccgggacg atttattgat
     4501 atattaacac taaatgatgg gaaattactt agtactaaaa gaataacgtt cgtagtaatg
     4561 gatgaggcag acaggctgtt cgatttaggt tttgaacctc aaataacgca aatcatgaaa
     4621 actgttcgac cggataaaca gtgtgttcta tttagtgcaa cttttccgaa caaactacgc
     4681 agttttgccg taagagtttt gcattcccca atatctatta cgattaattc aaagggaatg
     4741 gttaatgaaa acgtaaaaca aaagtttaga atatgccatt cagaagatga aaaatttgac
     4801 aatctggtgc agcttatcca tgagcgcagt gaattttttg acgaggttca aagtgaaaat
     4861 gacggacagt caagcgatgt tgaagaagtt gatgctaaag ccattatatt tgtatcaagt
     4921 cagaatattt gcgacttcat ttcaaaaaag ctgttaaatg ctggaattgt gacctgtgct
     4981 attcatgcag gtaagccata tcaagaaagg cttatgaact tagaaaagtt caaacgagaa
     5041 aagaacagta ttcttctctg tacagaggtt ctctcaagag gtttaaatgt tcccgaagtg
     5101 tcgttggtta ttatctataa cgccgtcaaa actttcgcgc aatatgttca tactactgga
     5161 agaactgcca gaggtagtcg ctccggtact gctattactc ttctattgca tgatgaactg
     5221 tcaggtgctt atatcttgag caaggcaatg cgcgatgaag aaataaaagc tttagaccct
     5281 ctgcaagcaa aagaacttca ggaaatgagt gccaagtttg aatccggtat gaaaaagggt
     5341 aagtttaggt tgtcgaaagg gttcggcggc aaaggactgg aaaatatcaa aagtaaaaga
     5401 gaggaagctc aaaataaaga tttggagctc aaaaagaatg ataagaggag cgatgacttg
     5461 gaaaagaaaa tcagcaatcc acgcgaaggg catgacagtg tgtctgagtc ttctgcgctt
     5521 attccgagac tcaattatga actattcaaa gagtctacag acgggtcgat catcttttac
     5581 gccaaggttt atattaatga tttaccccag attgtaagat gggaggctac aaagaataca
     5641 accttattat ttatcaagca tgaaaccgga tgtagtatta ctaacaaggg taaattttac
     5701 cctgaaggga aggaaccaaa gaatgaaaac gatgagccta aactatactt attgatcgaa
     5761 ggccaagatg agaaagacat acaattaagt atagaattgc tggagcaaaa agttaaagag
     5821 ggggtcgtaa aggctgcaag cttgtctttg aagagtacta aatactagtt tttagaattt
     5881 aactcactcg tggtgctata tactttcgta gttagttctg aaatttttag ggtggtaatt
     5941 taaagcatat tcaatcataa taaaaaaaaa aaaagagaat tattaggaaa aataaaaaca
     6001 aaataatata taatatatga tgtagaaaaa aatcttgaaa tttttatcga ttcgaataaa
     6061 atggaattta aaaaagtggg aaagaaacaa tgagaattca taggatataa tgaaaggtaa
     6121 atgctttaaa aaaaaattaa gaggggaaaa taaaagtaga aaagcaaata aaaacaaaac
     6181 atggaataaa agtaacttca aaaaaattta tcaagaaaat ggtaggcttg aagaatataa
     6241 taatagtttt ttagttgatt tgagatgtga tatgacaaat aaaaaataat gaaataacga
     6301 taaaaataag gaagggaaaa actgacaatc gaataataat attaataatg gtaagtttat
     6361 cacatcaatt cataggggga tcccccgggc tgcaggaatt cgatatcaag cttatcgata
     6421 ccgtcgacct cgaggggggg cccggtaccc aattcgccct atagtgagtc gtattacgcg
     6481 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     6541 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     6601 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcgacgc gccctgtagc
     6661 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     6721 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     6781 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     6841 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     6901 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     6961 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     7021 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     7081 aaaatattaa cgtttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta
     7141 tttcacaccg catagatccg tcgagttcaa gagaaaaaaa aagaaaaagc aaaaagaaaa
     7201 aaggaaagcg cgcctcgttc agaatgacac gtatagaatg atgcattacc ttgtcatctt
     7261 cagtatcata ctgttcgtat acatacttac tgacattcat aggtatacat atatacacat
     7321 gtatatatat cgtatgctgc agctttaaat aatcggtgtc actacataag aacacctttg
     7381 gtggagggaa catcgttggt accattgggc gaggtggctt ctcttatggc aaccgcaaga
     7441 gccttgaacg cactctcact acggtgatga tcattcttgc ctcgcagaca atcaacgtgg
     7501 agggtaattc tgctagcctc tgcaaagctt tcaagaaaat gcgggatcat ctcgcaagag
     7561 agatctccta ctttctccct ttgcaaacca agttcgacaa ctgcgtacgg cctgttcgaa
     7621 agatctacca ccgctctgga aagtgcctca tccaaaggcg caaatcctga tccaaacctt
     7681 tttactccac gcgccagtag ggcctcttta aaagcttgac cgagagcaat cccgcagtct
     7741 tcagtggtgt gatggtcgtc tatgtgtaag tcaccaatgc actcaacgat tagcgaccag
     7801 ccggaatgct tggccagagc atgtatcata tggtccagaa accctatacc tgtgtggacg
     7861 ttaatcactt gcgattgtgt ggcctgttct gctactgctt ctgcctcttt ttctgggaag
     7921 atcgagtgct ctatcgctag gggaccaccc tttaaagaga tcgcaatctg aatcttggtt
     7981 tcatttgtaa tacgctttac tagggctttc tgctctgtca tctttgcctt cgtttatctt
     8041 gcctgctcat tttttagtat attcttcgaa gaaatcacat tactttatat aatgtataat
     8101 tcattatgtg ataatgccaa tcgctaagaa aaaaaaagag tcatccgcta ggggaaaaaa
     8161 aaaaatgaaa atcattaccg aggcataaaa aaatatagag tgtactagag gaggccaaga
     8221 gtaatagaaa aagaaaattg cgggaaagga ctgtgttatg acttccctga ctaatgccgt
     8281 gttcaaacga tacctggcag tgactcctag cgctcaccaa gctcttaaaa cgggaattta
     8341 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     8401 ccaacacgcg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     8461 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     8521 gcga
//
