LOCUS       sequence-182552-        8127 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Encodes Hsh155 R294L D450G for expression in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8127)
  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 8127)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 R294L/D450G
FEATURES             Location/Qualifiers
     source          1..8127
                     /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        191..471
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             472..1146
                     /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(1249..1704)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1386..1407)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1598..1617
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1830..1852
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1844..1861
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1845..1861
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1871..1890
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1871..1889
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5354..5374)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5354..5372)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5393..5409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5393..5409)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5406..5428)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5417..5433
                     /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(5441..5447,5448..5465,5466..5471))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5486..5507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5624..5641)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5795..6383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5875..5894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6554..7345,7346..7414))
                     /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     7177..7196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7415..7519)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7556..8059
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8100..8118
                     /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 accatagatc acgacattac tatatatata atataggaag catttaatag aacagcatcg
      241 taatatatgt gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat
      301 tgaaaaatag cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga
      361 ttaagaatta attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga
      421 ctattgagca cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt
      481 attaatttca caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag
      541 gccgcagaat gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat
      601 agaaagagaa caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat
      661 aaaaatagtt caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag
      721 gatgttttgg ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag
      781 tcgtggcaag aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca
      841 aaagactgca acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg
      901 tttgattcag aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg
      961 gttggaaggc aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca
     1021 gaaaatgttg gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt
     1081 gtggagacaa atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag
     1141 aaataggtta ttactgagta gtatttattt aagtattgtt tgtgcacttg ccgatctatg
     1201 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg
     1261 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1321 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     1381 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     1441 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     1501 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1561 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     1621 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1681 ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg
     1741 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     1801 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     1861 tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc cccctcgagc
     1921 atgtactttc cgcctaacta ccatcacctg tctttattta ttccgctgac agatatgcac
     1981 aaccaattat cgtctcttga aagcttttta gacgtgattg ctgttatttg gcacgttgtt
     2041 agtatttaag cttattcggg tttgcattga tatttgcccg agcagaaatc tccttctcga
     2101 tgagttagaa ctgaaacatc tcttcaatta cttatacata ataaacatag aaagccagga
     2161 cctgtcatca tgagtcatcc gattcaattt gtaaacgcta ataatagcga taagagccac
     2221 caattaggcg gtcaatattc tattcctcag gatttgaggg aaaacctgca gaaagaagct
     2281 gcacgtattg gtgagaacga aaaagatgtg ctacaggaaa aaatggaaac taggacagtg
     2341 caaaatcgag aggattcata ccacaaaaga aggtttgata tgaagtttga gccggattcc
     2401 gatacacaaa ctgttacgtc ttcagaaaat acccaagacg ctgtcgtgcc aaggaaaaga
     2461 aaatctcgtt gggatgtaaa agggtacgag ccacctgatg aatcttcaac agcggtcaaa
     2521 gaaaactctg atagtgcttt ggttaatgta gaaggaatcc atgatttgat gttttttaaa
     2581 ccttctgacc ataaatattt cgccgacgtt atatccaaaa aacccataga tgagttaaat
     2641 aaggatgaaa agaaagaaag aaccctgtcg atgttacttc tgaaaataaa aaatggcaat
     2701 acagcgagta ggaggacttc tatgcggatt ttaaccgata aagctgttac atttggtcca
     2761 gaaatgatat ttaaccgctt attgcccata ttactagaca gaagtttgga agaccaggaa
     2821 agacatctaa tgatcaagac catagatcgc gtcctttacc agctaggaga tttaacaaaa
     2881 ccttacgttc acaaaatact tgtagtagct gccccattgc taatagatga agatccaatg
     2941 gttcgttcaa caggccagga gataatcacc aatctgtcta cagtcgctgg cctaaaaacc
     3001 attctaacag taatgagacc tgatattgag aatgaggatg agtatgttct aaatgtaact
     3061 tcaagagcgg cggctgtggt ggcaaaagcc cttggtgtta atcaattgct tcctttcatc
     3121 aatgcagcat gccactcacg gaagtcttgg aaagcaaggc atactggcat aaaaattgtg
     3181 cagcaaatag gtatactttt gggaataggt gtcttgaatc atcttactgg gctgatgagt
     3241 tgtatcaaag actgcctaat ggatgatcac gttccggtgc gaattgttac agcacatact
     3301 ttatcaacac tagcagaaaa ttcttatcca tatggtatcg aagttttcaa cgttgtactg
     3361 gagccattat ggaagggtat aagaagtcat cgtggtaaag tgctatcttc ttttttaaaa
     3421 gctgttggat ctatgatccc attgatggac ccagagtatg ccggttatta taccacagaa
     3481 gcaatgagaa taattagacg tgaattcgac tctcccggcg atgaaatgaa aaagaccatt
     3541 cttttagtat tacaaaaatg tagcgctgta gaatccataa caccaaaatt tctgagagaa
     3601 gaaatcgcgc cagaattctt tcaaaaattt tgggtcagac gagtggcatt ggatagacca
     3661 ttaaacaaag ttgttacata tactactgtt acattagcaa agaaattagg ttgttcttat
     3721 accattgata aattattaac accattgaga gatgaagctg agccgttcag aacaatggct
     3781 gtccatgctg ttactagaac agtaaactta ttagggacag cagatttaga tgagagatta
     3841 gaaacgaggc ttattgatgc gctcttgata gcatttcaag aacaaacaaa tagtgattct
     3901 ataatcttta aagggtttgg ggcagtgacg gtgtcacttg acatccgtat gaaaccgttt
     3961 ttagcaccca ttgtgagtac gattttaaat catttaaagc ataaaactcc cttggttcgt
     4021 caacatgcag cagatctatg tgccatacta ataccagtca tcaagaactg tcacgaattc
     4081 gaaatgttga ataaattgaa cattatattg tatgaatcct taggcgaagt atatccagag
     4141 gtattaggtt ctataatcaa cgctatgtac tgtattacta gcgtaatgga tttggataaa
     4201 ctacaacctc ctattaatca aattttacca accttaactc caattttaag aaacaagcat
     4261 agaaaagtcg aagtaaacac tatcaaattt gttggactta ttggtaaatt agcacctacc
     4321 tatgcacctc ctaaagaatg gatgagaata tgttttgaac tattagaact tttgaaaagt
     4381 acaaataaag aaataagaag atcagcaaat gccacgtttg gattcattgc tgaagcaatt
     4441 ggacctcacg atgttcttgt tgcgttattg aacaatttaa aggttcaaga acgtcagtta
     4501 cgtgtatgca cggctgttgc tatcggtatc gttgctaaag tgtgtggtcc ttacaacgtg
     4561 ctgccagtaa tcatgaatga atacacaaca ccagagacta atgttcaaaa cggtgttctc
     4621 aaggccatgt cattcatgtt tgagtacatt ggtaacatgt ctaaggatta catatacttc
     4681 ataacaccat tattagaaga tgcactcacc gatagggatt tagttcatcg tcagacagcg
     4741 tcgaacgtaa taacccactt agctttaaac tgttcaggta caggtcacga agatgccttc
     4801 attcatttaa tgaatcttct aatacccaat atttttgaaa cttcaccaca tgctattatg
     4861 cgtattttag aaggtttgga ggcactgagt caagctcttg gccctggact attcatgaac
     4921 tacatatggg caggattgtt ccatccagcg aagaatgtaa ggaaagcatt ttggagggtg
     4981 tataataaca tgtacgtaat gtaccaggac gccatggtac ctttttaccc cgttacacca
     5041 gacaacaatg aagaatatat agaagaactg gatttagttc tgtgaatata tatagagcat
     5101 ataaataacc acaacttgta agaatatttt acttgacaca cctatatcat cacgtcacga
     5161 tgccttcata gattatgatg aaatataaac agaacctgct attggaaaaa aaagggcagg
     5221 aaaaaatttt gtgaaagtga gtaaaagaaa gatgaagagc gagacatgga taataagaaa
     5281 tagatgtcac taatggacaa ttggaagact gatatggaaa gttacgatga aggaggagct
     5341 ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc
     5401 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5461 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5521 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5581 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5641 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5701 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5761 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5821 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5881 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5941 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6001 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6061 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6121 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6181 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6241 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6301 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6361 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6421 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6481 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6541 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6601 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6661 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6721 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6781 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6841 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6901 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6961 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7021 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7081 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7141 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7201 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7261 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7321 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7381 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7441 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7501 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgggtcc ttttcatcac
     7561 gtgctataaa aataattata atttaaattt tttaatataa atatataaat taaaaataga
     7621 aagtaaaaaa agaaattaaa gaaaaaatag tttttgtttt ccgaagatgt aaaagactct
     7681 agggggatcg ccaacaaata ctacctttta tcttgctctt cctgctctca ggtattaatg
     7741 ccgaattgtt tcatcttgtc tgtgtagaag accacacacg aaaatcctgt gattttacat
     7801 tttacttatc gttaatcgaa tgtatatcta tttaatctgc ttttcttgtc taataaatat
     7861 atatgtaaag tacgcttttt gttgaaattt tttaaacctt tgtttatttt tttttcttca
     7921 ttccgtaact cttctacctt ctttatttac tttctaaaat ccaaatacaa aacataaaaa
     7981 taaataaaca cagagtaaat tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt
     8041 aagttacagg caagcgatcc gtctaagaaa ccattattat catgacatta acctataaaa
     8101 ataggcgtat cacgaggccc tttcgtc
//
