LOCUS       sequence-189179-        8523 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 8523)
  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 8523)
  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..8523
                     /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    76..579
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        609..713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(714..782,783..1574)
                     /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(932..951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1745..2333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2234..2253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2487..2504
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2621..2642
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2657..2662,2663..2680,2681..2687)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2695..2711
                     /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     2700..2722
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2719..2735
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2719..2735
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2754..2774
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2756..2774
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6415..6431)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6416..6432)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(6456..6475)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6457..6475)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6485..6502)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6485..6501)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6494..6516)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      6642..7097
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6729..6748)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6939..6960
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7357..8019)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(8020..8207)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(8354..8373)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8473..8495
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagggtcc ttttcatcac gtgctataaa aataattata atttaaattt tttaatataa
      121 atatataaat taaaaataga aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt
      181 ccgaagatgt aaaagactct agggggatcg ccaacaaata ctacctttta tcttgctctt
      241 cctgctctca ggtattaatg ccgaattgtt tcatcttgtc tgtgtagaag accacacacg
      301 aaaatcctgt gattttacat tttacttatc gttaatcgaa tgtatatcta tttaatctgc
      361 ttttcttgtc taataaatat atatgtaaag tacgcttttt gttgaaattt tttaaacctt
      421 tgtttatttt tttttcttca ttccgtaact cttctacctt ctttatttac tttctaaaat
      481 ccaaatacaa aacataaaaa taaataaaca cagagtaaat tcccaaatta ttccatcatt
      541 aaaagatacg aggcgcgtgt aagttacagg caagcgatcc gtcaggtggc acttttcggg
      601 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      661 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      721 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      781 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
      841 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
      901 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
      961 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1021 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1081 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1141 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1201 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1261 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1321 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1381 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1441 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1501 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1561 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1621 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1681 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1741 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1801 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     1861 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     1921 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     1981 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2041 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2101 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2161 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2221 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2281 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2341 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2401 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2461 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2521 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2581 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2641 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2701 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag cgcgcaatta
     2761 accctcacta aagggaacaa aagctggagc tccaccgcgg tggcggccgc tctagaattt
     2821 aaacccggat tctgcctcaa tgggtagatc agacacctta gaagtaagga tagagctgct
     2881 ttgcaaaact ggtaatgatc cattaccatc accaggcgca ttggcagtaa ctttggagtc
     2941 tttttttgag actgtggacg attcatcacc agtaattgag ccgtactgac ggtcatagtc
     3001 ttcttgtgac atccatattg gcttctctgg tttggttgac attctacttt tctaagacgg
     3061 ataatggacg actccacgtt gtagtagttg tcacattaat cggaaaagtg tctcacatgt
     3121 aaaggattaa aataagaaat cttctaattt atgttgaaat atcttgattt accattaccc
     3181 ggattagatg aaactaatta aagtcattta gagcaatgga ccaaactaaa ggtattagag
     3241 ttctttctaa tataatacag cgccaaataa ttcgagaagg gaaatcataa cgctgttgcc
     3301 ttgggtttat ttgaggtatg gaaactattg attcgaagca aaatattaat agggagtctt
     3361 tattggagga aaggaggaaa aaactagcaa aatggaaaca aaaaaaagca caatttgatg
     3421 ctcagaaaga gcatcaaact tcacgcaatg acattgttac taacagttta gaaggtaagc
     3481 agactactga aaagtttacg gaaagacagg aacgtgtaaa agaagagctt cggaagagaa
     3541 aaaatgaatt taggaaatct gacgaacctg tatcggttaa gccttcgaag aaaaagtcga
     3601 agagaagtaa ggtaaagaag aaaatatcct tcgacttcag tgatgacgat gattctgaaa
     3661 taggagtttc ttttagatcg aaggagcaca ttcaaaaagc tccagagcat gataatgaaa
     3721 aagatccact ggatgagttt atgacgtcat tgaaggagga gaaaatgagt aatagtaaag
     3781 gcatgtacga cagaggcgac attcttgatg tagaggatca attgtttgaa cttggaggaa
     3841 ccgacgatga ggatgttgaa gataatacgg ataattctaa tatagcgaaa attgctaaac
     3901 tcaaagcaaa aaaacgtgta aagcaaattt attactctcc agaagagctt gagccgttcc
     3961 aaaaaaattt ttatatagaa tctgaaacag tttcctcgat gtcagagatg gaagttgagg
     4021 cacttagact tagtctggat aatatcaaaa taaagggaac aggttgcccc aaaccagtca
     4081 caaagtggtc tcaactggga ctatcaacgg acaccatggt tttaattaca gaaaagttgc
     4141 actttggctc cttaacgcct attcaatccc aggctcttcc cgctattatg tcaggtcgtg
     4201 atgttatagg aatatcaaaa actggctccg gcaagaccat ctcctatctt ttaccattgc
     4261 taagacaagt aaaagctcaa cggccattat caaaacatga aacagggccc atgggtttaa
     4321 ttctggctcc aactagagag ctagctttac aaatacatga agaagttact aaattcacag
     4381 aagcagatac atccattaga tcagtatgct gtacaggagg ttctgaaatg aaaaagcaga
     4441 ttactgatct taaaagaggc actgagattg ttgttgccac accgggacga tttattgata
     4501 tattaacact aaatgatggg aaattactta gtactaaaag aataacgttc gtagtaatgg
     4561 atgaggcaga caggctgttc gatttaggtt ttgaacctca aataacgcaa atcatgaaaa
     4621 ctgttcgacc ggataaacag tgtgttctat ttagtgcaac ttttccgaac aaactacgca
     4681 gttttgccgt aagagttttg cattccccaa tatctattac gattaattca aagggaatgg
     4741 ttaatgaaaa cgtaaaacaa aagtttagaa tatgccattc agaagatgaa aaatttgaca
     4801 atctggtgca gcttatccat gagcgcagtg aattttttga cgaggttcaa agtgaaaatg
     4861 acggacagtc aagcgatgtt gaagaagttg atgctaaagc cattatattt gtatcaagtc
     4921 agaatatttg cgacttcatt tcaaaaaagc tgttaaatgc tggaattgtg acctgtgcta
     4981 ttcatgcagg taagccatat caagaaaggc ttatgaactt agaaaagttc aaacgagaaa
     5041 agaacagtat tcttctctgt acagaggttc tctcaagagg tttaaatgtt cccgaagtgt
     5101 cgttggttat tatctataac gccgtcaaaa ctttcgcgca atatgttcat actactggaa
     5161 gaactgccag aggtagtcgc tccggtactg ctattactct tctattgcat gatgaactgt
     5221 caggtgctta tatcttgagc aaggcaatgc gcgatgaaga aataaaagct ttagaccctc
     5281 tgcaagcaaa agaacttcag gaaatgagtg ccaagtttga atccggtatg aaaaagggta
     5341 agtttaggtt gtcgaaaggg ttcggcggca aaggactgga aaatatcaaa agtaaaagag
     5401 aggaagctca aaataaagat ttggagctca aaaagaatga taagaggagc gatgacttgg
     5461 aaaagaaaat cagcaatcca cgcgaagggc atgacagtgt gtctgagtct tctgcgctta
     5521 ttccgagact caattatgaa ctattcaaag agtctacaga cgggtcgatc atcttttacg
     5581 ccaaggttta tattaatgat ttaccccaga ttgtaagatg ggaggctaca aagaatacaa
     5641 ccttattatt tatcaagcat gaaaccggat gtagtattac taacaagggt aaattttacc
     5701 ctgaagggaa ggaaccaaag aatgaaaacg atgagcctaa actatactta ttgatcgaag
     5761 gccaagatga gaaagacata caattaagta tagaattgct ggagcaaaaa gttaaagagg
     5821 gggtcgtaaa ggctgcaagc ttgtctttga agagtactaa atactagttt ttagaattta
     5881 actcactcgt ggtgctatat actttcgtag ttagttctga aatttttagg gtggtaattt
     5941 aaagcatatt caatcataat aaaaaaaaaa aaagagaatt attaggaaaa ataaaaacaa
     6001 aataatatat aatatatgat gtagaaaaaa atcttgaaat ttttatcgat tcgaataaaa
     6061 tggaatttaa aaaagtggga aagaaacaat gagaattcat aggatataat gaaaggtaaa
     6121 tgctttaaaa aaaaattaag aggggaaaat aaaagtagaa aagcaaataa aaacaaaaca
     6181 tggaataaaa gtaacttcaa aaaaatttat caagaaaatg gtaggcttga agaatataat
     6241 aatagttttt tagttgattt gagatgtgat atgacaaata aaaaataatg aaataacgat
     6301 aaaaataagg aagggaaaaa ctgacaatcg aataataata ttaataatgg taagtttatc
     6361 acatcaattc atagggggat cccccgggct gcaggaattc gatatcaagc ttatcgatac
     6421 cgtcgacctc gagggggggc ccggtaccca attcgcccta tagtgagtcg tattacgcgc
     6481 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     6541 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     6601 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     6661 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     6721 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     6781 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     6841 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     6901 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     6961 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     7021 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     7081 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     7141 caccgcatag atccgtcgag ttcaagagaa aaaaaaagaa aaagcaaaaa gaaaaaagga
     7201 aagcgcgcct cgttcagaat gacacgtata gaatgatgca ttaccttgtc atcttcagta
     7261 tcatactgtt cgtatacata cttactgaca ttcataggta tacatatata cacatgtata
     7321 tatatcgtat gctgcagctt taaataatcg gtgtcactac ataagaacac ctttggtgga
     7381 gggaacatcg ttggtaccat tgggcgaggt ggcttctctt atggcaaccg caagagcctt
     7441 gaacgcactc tcactacggt gatgatcatt cttgcctcgc agacaatcaa cgtggagggt
     7501 aattctgcta gcctctgcaa agctttcaag aaaatgcggg atcatctcgc aagagagatc
     7561 tcctactttc tccctttgca aaccaagttc gacaactgcg tacggcctgt tcgaaagatc
     7621 taccaccgct ctggaaagtg cctcatccaa aggcgcaaat cctgatccaa acctttttac
     7681 tccacgcacg gcccctaggg cctctttaaa agcttgaccg agagcaatcc cgcagtcttc
     7741 agtggtgtga tggtcgtcta tgtgtaagtc accaatgcac tcaacgatta gcgaccagcc
     7801 ggaatgcttg gccagagcat gtatcatatg gtccagaaac cctatacctg tgtggacgtt
     7861 aatcacttgc gattgtgtgg cctgttctgc tactgcttct gcctcttttt ctgggaagat
     7921 cgagtgctct atcgctaggg gaccaccctt taaagagatc gcaatctgaa tcttggtttc
     7981 atttgtaata cgctttacta gggctttctg ctctgtcatc tttgccttcg tttatcttgc
     8041 ctgctcattt tttagtatat tcttcgaaga aatcacatta ctttatataa tgtataattc
     8101 attatgtgat aatgccaatc gctaagaaaa aaaaagagtc atccgctagg tggaaaaaaa
     8161 aaaatgaaaa tcattaccga ggcataaaaa aatatagagt gtactagagg aggccaagag
     8221 taatagaaaa agaaaattgc gggaaaggac tgtgttatga cttccctgac taatgccgtg
     8281 ttcaaacgat acctggcagt gactcctagc gctcaccaag ctcttaaaac gggaatttat
     8341 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     8401 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     8461 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     8521 cga
//
