LOCUS       sequence-170294-        8124 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Hsh155 N295D H331D for expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8124)
  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 8124)
  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: pRS414 Hsh155 N295D/H331D
FEATURES             Location/Qualifiers
     source          1..8124
                     /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        187..467
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             468..1142
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(1241..1696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1378..1399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1590..1609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1826..1848
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1840..1857
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1841..1857
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1867..1886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1867..1885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5350..5370)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5350..5368)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5389..5405)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5389..5405)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5402..5424)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5413..5429
                     /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(5437..5443,5444..5461,5462..5467))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(5620..5637)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5791..6379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5871..5890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6550..7341,7342..7410))
                     /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     7173..7192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7411..7515)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7552..8055
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8097..8115
                     /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 accataaacg acattactat atatataata taggaagcat ttaatagaca gcatcgtaat
      241 atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt ctttattgaa
      301 aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt tgccgattaa
      361 gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat tggtgactat
      421 tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg tctgttatta
      481 atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc acagaggccg
      541 cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg cccaatagaa
      601 agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa gcatataaaa
      661 atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct aaggaggatg
      721 ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga gatgagtcgt
      781 ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta tttccaaaag
      841 actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt cccttgtttg
      901 attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct gactgggttg
      961 gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg acgccagaaa
     1021 atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc ggaggtgtgg
     1081 agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat gctaagaaat
     1141 aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgccta tgcggtgtga
     1201 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaaa cgttaatatt
     1261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     1321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     1381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     1441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     1501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     1561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     1621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     1681 ccgctacagg gcgcgtcgcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1741 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     1801 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgag
     1861 cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggccccccct cgagcatgta
     1921 ctttccgcct aactaccatc acctgtcttt atttattccg ctgacagata tgcacaacca
     1981 attatcgtct cttgaaagct ttttagacgt gattgctgtt atttggcacg ttgttagtat
     2041 ttaagcttat tcgggtttgc attgatattt gcccgagcag aaatctcctt ctcgatgagt
     2101 tagaactgaa acatctcttc aattacttat acataataaa catagaaagc caggacctgt
     2161 catcatgagt catccgattc aatttgtaaa cgctaataat agcgataaga gccaccaatt
     2221 aggcggtcaa tattctattc ctcaggattt gagggaaaac ctgcagaaag aagctgcacg
     2281 tattggtgag aacgaaaaag atgtgctaca ggaaaaaatg gaaactagga cagtgcaaaa
     2341 tcgagaggat tcataccaca aaagaaggtt tgatatgaag tttgagccgg attccgatac
     2401 acaaactgtt acgtcttcag aaaataccca agacgctgtc gtgccaagga aaagaaaatc
     2461 tcgttgggat gtaaaagggt acgagccacc tgatgaatct tcaacagcgg tcaaagaaaa
     2521 ctctgatagt gctttggtta atgtagaagg aatccatgat ttgatgtttt ttaaaccttc
     2581 tgaccataaa tatttcgccg acgttatatc caaaaaaccc atagatgagt taaataagga
     2641 tgaaaagaaa gaaagaaccc tgtcgatgtt acttctgaaa ataaaaaatg gcaatacagc
     2701 gagtaggagg acttctatgc ggattttaac cgataaagct gttacatttg gtccagaaat
     2761 gatatttaac cgcttattgc ccatattact agacagaagt ttggaagacc aggaaagaca
     2821 tctaatgatc aagaccatag atcgcgtcct ttaccagcta ggagatttaa caaaacctta
     2881 cgttcacaaa atacttgtag tagctgcccc attgctaata gatgaagatc caatggttcg
     2941 ttcaacaggc caggagataa tcaccaatct gtctacagtc gctggcctaa aaaccattct
     3001 aacagtaatg agacctgata ttgagaatga ggatgagtat gttagagatg taacttcaag
     3061 agcggcggct gtggtggcaa aagcccttgg tgttaatcaa ttgcttcctt tcatcaatgc
     3121 agcatgccac tcacggaagt cttggaaagc aagggatact ggcataaaaa ttgtgcagca
     3181 aataggtata cttttgggaa taggtgtctt gaatcatctt actgggctga tgagttgtat
     3241 caaagactgc ctaatggatg atcacgttcc ggtgcgaatt gttacagcac atactttatc
     3301 aacactagca gaaaattctt atccatatgg tatcgaagtt ttcaacgttg tactggagcc
     3361 attatggaag ggtataagaa gtcatcgtgg taaagtgcta tcttcttttt taaaagctgt
     3421 tggatctatg atcccattga tggacccaga gtatgccggt tattatacca cagaagcaat
     3481 gagaataatt agacgtgaat tcgactctcc cgatgatgaa atgaaaaaga ccattctttt
     3541 agtattacaa aaatgtagcg ctgtagaatc cataacgcca aaatttctga gagaagaaat
     3601 cgcgccagaa ttctttcaaa aattttgggt cagacgagtg gcattggata gaccattaaa
     3661 caaagttgtt acatatacta ctgttacatt agcaaagaaa ttaggttgtt cttataccat
     3721 tgataaatta ttaacaccat tgagagatga agctgagccg ttcagaacaa tggctgtcca
     3781 tgctgttact agaacagtaa acttattagg gacagcagat ttagatgaga gattagaaac
     3841 gaggcttatt gatgcgctct tgatagcatt tcaagaacaa acaaatagtg attctataat
     3901 ctttaaaggg tttggggcag tgacggtgtc acttgacatc cgtatgaaac cgtttttagc
     3961 acccattgtg agtacgattt taaatcattt aaagcataaa actcccttgg ttcgtcaaca
     4021 tgcagcagat ctatgtgcca tactaatacc agtcatcaag aactgtcacg aattcgaaat
     4081 gttgaataaa ttgaacatta tattgtatga atccttaggc gaagtatatc cagaggtatt
     4141 aggttctata atcaacgcta tgtactgtat tactagcgta atggatttgg ataaactaca
     4201 acctcctatt aatcaaattt taccaacctt aactccaatt ttaagaaaca agcatagaaa
     4261 agtcgaagta aacactatca aatttgttgg acttattggt aaattagcac ctacctatgc
     4321 acctcctaaa gaatggatga gaatatgttt tgaactatta gaacttttga aaagtacaaa
     4381 taaagaaata agaagatcag caaatgccac gtttggattc attgctgaag caattggacc
     4441 tcacgatgtt cttgttgcgt tattgaacaa tttaaaggtt caagaacgtc agttacgtgt
     4501 atgcacggct gttgctatcg gtatcgttgc taaagtgtgt ggtccttaca acgtgctgcc
     4561 agtaatcatg aatgaataca caacaccaga gactaatgtt caaaacggtg ttctcaaggc
     4621 catgtcattc atgtttgagt acattggtaa catgtctaag gattacatat acttcataac
     4681 accattatta gaagatgcac tcaccgatag ggatttagtt catcgtcaga cagcgtcgaa
     4741 cgtaataacc cacttagctt taaactgttc aggtacaggt cacgaagatg ccttcattca
     4801 tttaatgaat cttctaatac ccaatatttt tgaaacttca ccacatgcta ttatgcgtat
     4861 tttagaaggt ttggaggcac tgagtcaagc tcttggccct ggactattca tgaactacat
     4921 atgggcagga ttgttccatc cagcgaagaa tgtaaggaaa gcattttgga gggtgtataa
     4981 taacatgtac gtaatgtacc aggacgccat ggtacctttt taccccgtta caccagacaa
     5041 caatgaagaa tatatagaag aactggattt agttctgtga atatatatag agcatataaa
     5101 taaccacaac ttgtaagaat attttacttg acacacctat atcatcacgt cacgatgcct
     5161 tcatagatta tgatgaaata taaacagaac ctgctattgg aaaaaaaagg gcaggaaaaa
     5221 attttgtgaa agtgagtaaa agaaagatga agagcgagac atggataata agaaatagat
     5281 gtcactaatg gacaattgga agactgatat ggaaagttac gatgaaggag gagctcccag
     5341 cttttgttcc ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt
     5401 tcctgtgtga aattgttatc cgctcacaat tccacacaac ataggagccg gaagcataaa
     5461 gtgtaaagcc tggggtgcct aatgagtgag gtaactcaca ttaattgcgt tgcgctcact
     5521 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5581 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5641 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5701 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5761 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5821 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5881 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5941 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6001 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6061 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6121 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6181 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     6241 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6301 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6361 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6421 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6481 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6541 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6601 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6661 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6721 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6781 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6841 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6901 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6961 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7021 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7081 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7141 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7201 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7261 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7321 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7381 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     7441 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7501 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gggtcctttt catcacgtgc
     7561 tataaaaata attataattt aaatttttta atataaatat ataaattaaa aatagaaagt
     7621 aaaaaaagaa attaaagaaa aaatagtttt tgttttccga agatgtaaaa gactctaggg
     7681 ggatcgccaa caaatactac cttttatctt gctcttcctg ctctcaggta ttaatgccga
     7741 attgtttcat cttgtctgtg tagaagacca cacacgaaaa tcctgtgatt ttacatttta
     7801 cttatcgtta atcgaatgta tatctattta atctgctttt cttgtctaat aaatatatat
     7861 gtaaagtacg ctttttgttg aaatttttta aacctttgtt tatttttttt tcttcattcc
     7921 gtaactcttc taccttcttt atttactttc taaaatccaa atacaaaaca taaaaataaa
     7981 taaacacaga gtaaattccc aaattattcc atcattaaaa gatacgaggc gcgtgtaagt
     8041 tacaggcaag cgatccgtcc taagaaacca ttattatcat gacattaacc tataaaaata
     8101 ggcgtatcac gaggcccttt cgtc
//
