LOCUS       sequence-213542-        8127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This is a shuffle plasmid containing the HSH155 gene and the 
            surrounding genomic regions. It bears a TRP marker for shuffle into 
            a HSH155 deletion strain. It has the K227A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8127)
  AUTHORS   Carrocci TJ, Paulson JC, Hoskins AA
  TITLE     Functional Analysis of Hsh155/SF3b1 Interactions with the U2 
            snRNA/Branch Site Duplex.
  JOURNAL   RNA. 2018 May 11. pii: rna.065664.118. doi: 10.1261/rna.065664.118.
  PUBMED    29752352
REFERENCE   2  (bases 1 to 8127)
  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: "RNA. 2018 
            May 11. pii: rna.065664.118. doi: 10.1261/rna.065664.118."
COMMENT     Sequence Label: Hsh155 K227A
FEATURES             Location/Qualifiers
     source          1..8127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3233..3253)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3233..3251)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3272..3288)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3272..3288)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3285..3307)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3296..3312
                     /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(3320..3326,3327..3344,3345..3350))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3365..3386
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3503..3520)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3674..4262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3754..3773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4433..5224,5225..5293))
                     /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     5056..5075
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5294..5398)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5435..5938
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     5979..5997
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6035..6057)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6157..6176
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6197..6477
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6478..7152
                     /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(7255..7710)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7392..7413)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7604..7623
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7836..7858
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7850..7867
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7851..7867
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7877..7896
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7877..7895
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cttcaattac ttatacataa taaacataga aagccaggac ctgtcatcat gagtcatccg
       61 attcaatttg taaacgctaa taatagcgat aagagccacc aattaggcgg tcaatattct
      121 attcctcagg atttgaggga aaacctgcag aaagaagctg cacgtattgg tgagaacgaa
      181 aaagatgtgc tacaggaaaa aatggaaact aggacagtgc aaaatcgaga ggattcatac
      241 cacaaaagaa ggtttgatat gaagtttgag ccggattccg atacacaaac tgttacgtct
      301 tcagaaaata cccaagacgc tgtcgtgcca aggaaaagaa aatctcgttg ggatgtaaaa
      361 gggtacgagc cacctgatga atcttcaaca gcggtcaaag aaaactctga tagtgctttg
      421 gttaatgtag aaggaatcca tgatttgatg ttttttaaac cttctgacca taaatatttc
      481 gccgacgtta tatccaaaaa acccatagat gagttaaata aggatgaaaa gaaagaaaga
      541 accctgtcga tgttacttct gaaaataaaa aatggcaata cagcgagtag gaggacttct
      601 atgcggattt taaccgataa agctgttaca tttggtccag aaatgatatt taaccgctta
      661 ttgcccatat tactagacag aagtttggaa gaccaggaaa gacatctaat gatcaagacc
      721 atagatgctg tcctttacca gctaggagat ttaacaaaac cttacgttca caaaatactt
      781 gtagtagctg ccccattgct aatagatgaa gatccaatgg ttcgttcaac aggccaggag
      841 ataatcacca atctgtctac agtcgctggc ctaaaaacca ttctaacagt aatgagacct
      901 gatattgaga atgaggatga gtatgttaga aatgtaactt caagagcggc ggctgtggtg
      961 gcaaaagccc ttggtgttaa tcaattgctt cctttcatca atgcagcatg ccactcacgg
     1021 aagtcttgga aagcaaggca tactggcata aaaattgtgc agcaaatagg tatacttttg
     1081 ggaataggtg tcttgaatca tcttactggg ctgatgagtt gtatcaaaga ctgcctaatg
     1141 gatgatcacg ttccggtgcg aattgttaca gcacatactt tatcaacact agcagaaaat
     1201 tcttatccat atggtatcga agttttcaac gttgtactgg agccattatg gaagggtata
     1261 agaagtcatc gtggtaaagt gctatcttct tttttaaaag ctgttggatc tatgatccca
     1321 ttgatggacc cagagtatgc cggttattat accacagaag caatgagaat aattagacgt
     1381 gaattcgact ctcccgatga tgaaatgaaa aagaccattc ttttagtatt acaaaaatgt
     1441 agcgctgtag aatccataac gccaaaattt ctgagagaag aaatcgcgcc agaattcttt
     1501 caaaaatttt gggtcagacg agtggcattg gatagaccat taaacaaagt tgttacatat
     1561 actactgtta cattagcaaa gaaattaggt tgttcttata ccattgataa attattaaca
     1621 ccattgagag atgaagctga gccgttcaga acaatggctg tccatgctgt tactagaaca
     1681 gtaaacttat tagggacagc agatttagat gagagattag aaacgaggct tattgatgcg
     1741 ctcttgatag catttcaaga acaaacaaat agtgattcta taatctttaa agggtttggg
     1801 gcagtgacgg tgtcacttga catccgtatg aaaccgtttt tagcacccat tgtgagtacg
     1861 attttaaatc atttaaagca taaaactccc ttggttcgtc aacatgcagc agatctatgt
     1921 gccatactaa taccagtcat caagaactgt cacgaattcg aaatgttgaa taaattgaac
     1981 attatattgt atgaatcctt aggcgaagta tatccagagg tattaggttc tataatcaac
     2041 gctatgtact gtattactag cgtaatggat ttggataaac tacaacctcc tattaatcaa
     2101 attttaccaa ccttaactcc aattttaaga aacaagcata gaaaagtcga agtaaacact
     2161 atcaaatttg ttggacttat tggtaaatta gcacctacct atgcacctcc taaagaatgg
     2221 atgagaatat gttttgaact attagaactt ttgaaaagta caaataaaga aataagaaga
     2281 tcagcaaatg ccacgtttgg attcattgct gaagcaattg gacctcacga tgttcttgtt
     2341 gcgttattga acaatttaaa ggttcaagaa cgtcagttac gtgtatgcac ggctgttgct
     2401 atcggtatcg ttgctaaagt gtgtggtcct tacaacgtgc tgccagtaat catgaatgaa
     2461 tacacaacac cagagactaa tgttcaaaac ggtgttctca aggccatgtc attcatgttt
     2521 gagtacattg gtaacatgtc taaggattac atatacttca taacaccatt attagaagat
     2581 gcactcaccg atagggattt agttcatcgt cagacagcgt cgaacgtaat aacccactta
     2641 gctttaaact gttcaggtac aggtcacgaa gatgccttca ttcatttaat gaatcttcta
     2701 atacccaata tttttgaaac ttcaccacat gctattatgc gtattttaga aggtttggag
     2761 gcactgagtc aagctcttgg ccctggacta ttcatgaact acatatgggc aggattgttc
     2821 catccagcga agaatgtaag gaaagcattt tggagggtgt ataataacat gtacgtaatg
     2881 taccaggacg ccatggtacc tttttacccc gttacaccag acaacaatga agaatatata
     2941 gaagaactgg atttagttct gtgaatatat atagagcata taaataacca caacttgtaa
     3001 gaatatttta cttgacacac ctatatcatc acgtcacgat gccttcatag attatgatga
     3061 aatataaaca gaacctgcta ttggaaaaaa aagggcagga aaaaattttg tgaaagtgag
     3121 taaaagaaag atgaagagcg agacatggat aataagaaat agatgtcact aatggacaat
     3181 tggaagactg atatggaaag ttacgatgaa ggaggagctc cagcttttgt tccctttagt
     3241 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3301 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3361 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3421 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3481 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3541 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3601 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     3661 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     3721 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     3781 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     3841 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     3901 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     3961 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4021 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4081 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4141 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4201 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4261 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4321 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4381 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4441 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4501 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4561 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4621 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     4681 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     4741 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     4801 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     4861 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     4921 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     4981 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5041 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5101 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5161 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5221 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5281 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5341 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5401 catttccccg aaaagtgcca cctgggtcct tttcatcacg tgctataaaa ataattataa
     5461 tttaaatttt ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag
     5521 aaaaaatagt ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac
     5581 taccttttat cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct
     5641 gtgtagaaga ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat
     5701 gtatatctat ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg
     5761 ttgaaatttt ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc
     5821 tttatttact ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt
     5881 cccaaattat tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg
     5941 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     6001 ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga
     6061 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     6121 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     6181 gagtgcacca tagatcacga cattactata tatataatat aggaagcatt taatagaaca
     6241 gcatcgtaat atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt
     6301 ctttattgaa aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt
     6361 tgccgattaa gaattaattc ggtcgaaaaa agaaaaggag agggccaaga gggagggcat
     6421 tggtgactat tgagcacgtg agtatacgtg attaagcaca caaaggcagc ttggagtatg
     6481 tctgttatta atttcacagg tagttctggt ccattggtga aagtttgcgg cttgcagagc
     6541 acagaggccg cagaatgtgc tctagattcc gatgctgact tgctgggtat tatatgtgtg
     6601 cccaatagaa agagaacaat tgacccggtt attgcaagga aaatttcaag tcttgtaaaa
     6661 gcatataaaa atagttcagg cactccgaaa tacttggttg gcgtgtttcg taatcaacct
     6721 aaggaggatg ttttggctct ggtcaatgat tacggcattg atatcgtcca actgcatgga
     6781 gatgagtcgt ggcaagaata ccaagagttc ctcggtttgc cagttattaa aagactcgta
     6841 tttccaaaag actgcaacat actactcagt gcagcttcac agaaacctca ttcgtttatt
     6901 cccttgtttg attcagaagc aggtgggaca ggtgaacttt tggattggaa ctcgatttct
     6961 gactgggttg gaaggcaaga gagccccgaa agcttacatt ttatgttagc tggtggactg
     7021 acgccagaaa atgttggtga tgcgcttaga ttaaatggcg ttattggtgt tgatgtaagc
     7081 ggaggtgtgg agacaaatgg tgtaaaagac tctaacaaaa tagcaaattt cgtcaaaaat
     7141 gctaagaaat aggttattac tgagtagtat ttatttaagt attgtttgtg cacttgccga
     7201 tctatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggaaattg
     7261 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     7321 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     7381 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     7441 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     7501 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     7561 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     7621 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     7681 ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca ttcaggctgc gcaactgttg
     7741 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
     7801 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
     7861 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg ggcccccccc
     7921 tcgagcatgt actttccgcc taactaccat cacctgtctt tatttattcc gctgacagat
     7981 atgcacaacc aattatcgtc tcttgaaagc tttttagacg tgattgctgt tatttggcac
     8041 gttgttagta tttaagctta ttcgggtttg cattgatatt tgcccgagca gaaatctcct
     8101 tctcgatgag ttagaactga aacatct
//
