LOCUS       sequence-183092-        8523 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Prp5 WT 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 WT
FEATURES             Location/Qualifiers
     source          1..8523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        317..504
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             505..1167
                     /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"
     rep_origin      complement(1427..1882)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1564..1585)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1776..1795
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2008..2030
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2022..2039
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2023..2039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2049..2068
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2049..2067
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     2092..2108
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2093..2109
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5750..5770)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5750..5768)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5789..5805)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5789..5805)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5802..5824)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5813..5829
                     /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(5837..5843,5844..5861,5862..5867))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5882..5903
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6020..6037)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6191..6779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6271..6290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6950..7741,7742..7810))
                     /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     7573..7592
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7811..7915)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7945..8448
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8496..8514
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataaatt cccgttttaa gagcttggtg agcgctagga gtcactgcca ggtatcgttt
      241 gaacacggca ttagtcaggg aagtcataac acagtccttt cccgcaattt tctttttcta
      301 ttactcttgg cctcctctag tacactctat atttttttat gcctcggtaa tgattttcat
      361 tttttttttt ccacctagcg gatgactctt tttttttctt agcgattggc attatcacat
      421 aatgaattat acattatata aagtaatgtg atttcttcga agaatatact aaaaaatgag
      481 caggcaagat aaacgaaggc aaagatgaca gagcagaaag ccctagtaaa gcgtattaca
      541 aatgaaacca agattcagat tgcgatctct ttaaagggtg gtcccctagc gatagagcac
      601 tcgatcttcc cagaaaaaga ggcagaagca gtagcagaac aggccacaca atcgcaagtg
      661 attaacgtcc acacaggtat agggtttctg gaccatatga tacatgctct ggccaagcat
      721 tccggctggt cgctaatcgt tgagtgcatt ggtgacttac acatagacga ccatcacacc
      781 actgaagact gcgggattgc tctcggtcaa gcttttaaag aggccctagg ggccgtgcgt
      841 ggagtaaaaa ggtttggatc aggatttgcg cctttggatg aggcactttc cagagcggtg
      901 gtagatcttt cgaacaggcc gtacgcagtt gtcgaacttg gtttgcaaag ggagaaagta
      961 ggagatctct cttgcgagat gatcccgcat tttcttgaaa gctttgcaga ggctagcaga
     1021 attaccctcc acgttgattg tctgcgaggc aagaatgatc atcaccgtag tgagagtgcg
     1081 ttcaaggctc ttgcggttgc cataagagaa gccacctcgc ccaatggtac caacgatgtt
     1141 ccctccacca aaggtgttct tatgtagtga caccgattat ttaaagctgc agcatacgat
     1201 atatatacat gtgtatatat gtatacctat gaatgtcagt aagtatgtat acgaacagta
     1261 tgatactgaa gatgacaagg taatgcatca ttctatacgt gtcattctga acgaggcgcg
     1321 ctttcctttt ttctttttgc tttttctttt tttttctctt gaactcgacg gatctatgcg
     1381 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt
     1441 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     1501 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     1561 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     1621 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     1681 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     1741 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     1801 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     1861 aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc
     1921 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     1981 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2041 agcgcgcgta atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcg
     2101 acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccccc tatgaattga
     2161 tgtgataaac ttaccattat taatattatt attcgattgt cagtttttcc cttccttatt
     2221 tttatcgtta tttcattatt ttttatttgt catatcacat ctcaaatcaa ctaaaaaact
     2281 attattatat tcttcaagcc taccattttc ttgataaatt tttttgaagt tacttttatt
     2341 ccatgttttg tttttatttg cttttctact tttattttcc cctcttaatt tttttttaaa
     2401 gcatttacct ttcattatat cctatgaatt ctcattgttt ctttcccact tttttaaatt
     2461 ccattttatt cgaatcgata aaaatttcaa gatttttttc tacatcatat attatatatt
     2521 attttgtttt tatttttcct aataattctc tttttttttt tttattatga ttgaatatgc
     2581 tttaaattac caccctaaaa atttcagaac taactacgaa agtatatagc accacgagtg
     2641 agttaaattc taaaaactag tatttagtac tcttcaaaga caagcttgca gcctttacga
     2701 ccccctcttt aactttttgc tccagcaatt ctatacttaa ttgtatgtct ttctcatctt
     2761 ggccttcgat caataagtat agtttaggct catcgttttc attctttggt tccttccctt
     2821 cagggtaaaa tttacccttg ttagtaatac tacatccggt ttcatgcttg ataaataata
     2881 aggttgtatt ctttgtagcc tcccatctta caatctgggg taaatcatta atataaacct
     2941 tggcgtaaaa gatgatcgac ccgtctgtag actctttgaa tagttcataa ttgagtctcg
     3001 gaataagcgc agaagactca gacacactgt catgcccttc gcgtggattg ctgattttct
     3061 tttccaagtc atcgctcctc ttatcattct ttttgagctc caaatcttta ttttgagctt
     3121 cctctctttt acttttgata ttttccagtc ctttgccgcc gaaccctttc gacaacctaa
     3181 acttaccctt tttcataccg gattcaaact tggcactcat ttcctgaagt tcttttgctt
     3241 gcagagggtc taaagctttt atttcttcat cgcgcattgc cttgctcaag atataagcac
     3301 ctgacagttc atcatgcaat agaagagtaa tagcagtacc ggagcgacta cctctggcag
     3361 ttcttccagt agtatgaaca tattgcgcga aagttttgac ggcgttatag ataataacca
     3421 acgacacttc gggaacattt aaacctcttg agagaacctc tgtacagaga agaatactgt
     3481 tcttttctcg tttgaacttt tctaagttca taagcctttc ttgatatggc ttacctgcat
     3541 gaatagcaca ggtcacaatt ccagcattta acagcttttt tgaaatgaag tcgcaaatat
     3601 tctgacttga tacaaatata atggctttag catcaacttc ttcaacatcg cttgactgtc
     3661 cgtcattttc actttgaacc tcgtcaaaaa attcactgcg ctcatggata agctgcacca
     3721 gattgtcaaa tttttcatct tctgaatggc atattctaaa cttttgtttt acgttttcat
     3781 taaccattcc ctttgaatta atcgtaatag atattgggga atgcaaaact cttacggcaa
     3841 aactgcgtag tttgttcgga aaagttgcac taaatagaac acactgttta tccggtcgaa
     3901 cagttttcat gatttgcgtt atttgaggtt caaaacctaa atcgaacagc ctgtctgcct
     3961 catccattac tacgaacgtt attcttttag tactaagtaa tttcccatca tttagtgtta
     4021 atatatcaat aaatcgtccc ggtgtggcaa caacaatctc agtgcctctt ttaagatcag
     4081 taatctgctt tttcatttca gaacctcctg tacagcatac tgatctaatg gatgtatctg
     4141 cttctgtgaa tttagtaact tcttcatgta tttgtaaagc tagctctcta gttggagcca
     4201 gaattaaacc catgggccct gtttcatgtt ttgataatgg ccgttgagct tttacttgtc
     4261 ttagcaatgg taaaagatag gagatggtct tgccggagcc agtttttgat attcctataa
     4321 catcacgacc tgacataata gcgggaagag cctgggattg aataggcgtt aaggagccaa
     4381 agtgcaactt ttctgtaatt aaaaccatgg tgtccgttga tagtcccagt tgagaccact
     4441 ttgtgactgg tttggggcaa cctgttccct ttattttgat attatccaga ctaagtctaa
     4501 gctcctcaac ttccatctct gacatcgagg aaactgtttc agattctata taaaaatttt
     4561 tttggaacgg ctcaagctct tctggagagt aataaatttg ctttacacgt ttttttgctt
     4621 tgagtttagc aattttcgct atattagaat tatccgtatt atcttcaaca tcctcatcgt
     4681 cggttcctcc aagttcaaac aattgatcct ctacatcaag aatgtcgcct ctgtcgtaca
     4741 tgcctttact attactcatt ttctcctcct tcaatgacgt cataaactca tccagtggat
     4801 ctttttcatt atcatgctct ggagcttttt gaatgtgctc cttcgatcta aaagaaactc
     4861 ctatttcaga atcatcgtca tcactgaagt cgaaggatat tttcttcttt accttacttc
     4921 tcttcgactt tttcttcgaa ggcttaaccg atacaggttc gtcagatttc ctaaattcat
     4981 tttttctctt ccgaagctct tcttttacac gttcctgtct ttccgtaaac ttttcagtag
     5041 tctgcttacc ttctaaactg ttagtaacaa tgtcattgcg tgaagtttga tgctctttct
     5101 gagcatcaaa ttgtgctttt ttttgtttcc attttgctag ttttttcctc ctttcctcca
     5161 ataaagactc cctattaata ttttgcttcg aatcaatagt ttccatacct caaataaacc
     5221 caaggcaaca gcgttatgat ttcccttctc gaattatttg gcgctgtatt atattagaaa
     5281 gaactctaat acctttagtt tggtccattg ctctaaatga ctttaattag tttcatctaa
     5341 tccgggtaat ggtaaatcaa gatatttcaa cataaattag aagatttctt attttaatcc
     5401 tttacatgtg agacactttt ccgattaatg tgacaactac tacaacgtgg agtcgtccat
     5461 tatccgtctt agaaaagtag aatgtcaacc aaaccagaga agccaatatg gatgtcacaa
     5521 gaagactatg accgtcagta cggctcaatt actggtgatg aatcgtccac agtctcaaaa
     5581 aaagactcca aagttactgc caatgcgcct ggtgatggta atggatcatt accagttttg
     5641 caaagcagct ctatccttac ttctaaggtg tctgatctac ccattgaggc agaatccggg
     5701 tttaaattct agagcggccg ccaccgcggt ggagctccag cttttgttcc ctttagtgag
     5761 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5821 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5881 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5941 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6001 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6061 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6121 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6181 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6241 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6301 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6361 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6421 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6481 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6541 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6601 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6661 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6721 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6781 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6841 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6901 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6961 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7021 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7081 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7141 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7201 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7261 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7321 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7381 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7441 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7501 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7561 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7621 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7681 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7741 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7801 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7861 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7921 ttccccgaaa agtgccacct gacggatcgc ttgcctgtaa cttacacgcg cctcgtatct
     7981 tttaatgatg gaataatttg ggaatttact ctgtgtttat ttatttttat gttttgtatt
     8041 tggattttag aaagtaaata aagaaggtag aagagttacg gaatgaagaa aaaaaaataa
     8101 acaaaggttt aaaaaatttc aacaaaaagc gtactttaca tatatattta ttagacaaga
     8161 aaagcagatt aaatagatat acattcgatt aacgataagt aaaatgtaaa atcacaggat
     8221 tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt cggcattaat acctgagagc
     8281 aggaagagca agataaaagg tagtatttgt tggcgatccc cctagagtct tttacatctt
     8341 cggaaaacaa aaactatttt ttctttaatt tcttttttta ctttctattt ttaatttata
     8401 tatttatatt aaaaaattta aattataatt atttttatag cacgtgatga aaaggaccct
     8461 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     8521 gtc
//
