LOCUS       sequence-213584-        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 N747A 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 N747A
FEATURES             Location/Qualifiers
     source          1..8127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     10..29
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        10..28
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3493..3513)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3493..3511)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3532..3548)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3532..3548)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3545..3567)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3556..3572
                     /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(3580..3586,3587..3604,3605..3610))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3625..3646
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3763..3780)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3934..4522)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4014..4033)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4693..5484,5485..5553))
                     /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     5316..5335
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5554..5658)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5695..6198
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6239..6257
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6295..6317)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6417..6436
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6457..6737
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6738..7412
                     /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(7515..7970)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7652..7673)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7864..7883
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8096..8118
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8110..8127
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8111..8127
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gagcgcgcgt aatacgactc actatagggc gaattgggta ccgggccccc ccctcgagca
       61 tgtactttcc gcctaactac catcacctgt ctttatttat tccgctgaca gatatgcaca
      121 accaattatc gtctcttgaa agctttttag acgtgattgc tgttatttgg cacgttgtta
      181 gtatttaagc ttattcgggt ttgcattgat atttgcccga gcagaaatct ccttctcgat
      241 gagttagaac tgaaacatct cttcaattac ttatacataa taaacataga aagccaggac
      301 ctgtcatcat gagtcatccg attcaatttg taaacgctaa taatagcgat aagagccacc
      361 aattaggcgg tcaatattct attcctcagg atttgaggga aaacctgcag aaagaagctg
      421 cacgtattgg tgagaacgaa aaagatgtgc tacaggaaaa aatggaaact aggacagtgc
      481 aaaatcgaga ggattcatac cacaaaagaa ggtttgatat gaagtttgag ccggattccg
      541 atacacaaac tgttacgtct tcagaaaata cccaagacgc tgtcgtgcca aggaaaagaa
      601 aatctcgttg ggatgtaaaa gggtacgagc cacctgatga atcttcaaca gcggtcaaag
      661 aaaactctga tagtgctttg gttaatgtag aaggaatcca tgatttgatg ttttttaaac
      721 cttctgacca taaatatttc gccgacgtta tatccaaaaa acccatagat gagttaaata
      781 aggatgaaaa gaaagaaaga accctgtcga tgttacttct gaaaataaaa aatggcaata
      841 cagcgagtag gaggacttct atgcggattt taaccgataa agctgttaca tttggtccag
      901 aaatgatatt taaccgctta ttgcccatat tactagacag aagtttggaa gaccaggaaa
      961 gacatctaat gatcaagacc atagatcgcg tcctttacca gctaggagat ttaacaaaac
     1021 cttacgttca caaaatactt gtagtagctg ccccattgct aatagatgaa gatccaatgg
     1081 ttcgttcaac aggccaggag ataatcacca atctgtctac agtcgctggc ctaaaaacca
     1141 ttctaacagt aatgagacct gatattgaga atgaggatga gtatgttaga aatgtaactt
     1201 caagagcggc ggctgtggtg gcaaaagccc ttggtgttaa tcaattgctt cctttcatca
     1261 atgcagcatg ccactcacgg aagtcttgga aagcaaggca tactggcata aaaattgtgc
     1321 agcaaatagg tatacttttg ggaataggtg tcttgaatca tcttactggg ctgatgagtt
     1381 gtatcaaaga ctgcctaatg gatgatcacg ttccggtgcg aattgttaca gcacatactt
     1441 tatcaacact agcagaaaat tcttatccat atggtatcga agttttcaac gttgtactgg
     1501 agccattatg gaagggtata agaagtcatc gtggtaaagt gctatcttct tttttaaaag
     1561 ctgttggatc tatgatccca ttgatggacc cagagtatgc cggttattat accacagaag
     1621 caatgagaat aattagacgt gaattcgact ctcccgatga tgaaatgaaa aagaccattc
     1681 ttttagtatt acaaaaatgt agcgctgtag aatccataac gccaaaattt ctgagagaag
     1741 aaatcgcgcc agaattcttt caaaaatttt gggtcagacg agtggcattg gatagaccat
     1801 taaacaaagt tgttacatat actactgtta cattagcaaa gaaattaggt tgttcttata
     1861 ccattgataa attattaaca ccattgagag atgaagctga gccgttcaga acaatggctg
     1921 tccatgctgt tactagaaca gtaaacttat tagggacagc agatttagat gagagattag
     1981 aaacgaggct tattgatgcg ctcttgatag catttcaaga acaaacaaat agtgattcta
     2041 taatctttaa agggtttggg gcagtgacgg tgtcacttga catccgtatg aaaccgtttt
     2101 tagcacccat tgtgagtacg attttaaatc atttaaagca taaaactccc ttggttcgtc
     2161 aacatgcagc agatctatgt gccatactaa taccagtcat caagaactgt cacgaattcg
     2221 aaatgttgaa taaattgaac attatattgt atgaatcctt aggcgaagta tatccagagg
     2281 tattaggttc tataatcaac gctatgtact gtattactag cgtaatggat ttggataaac
     2341 tacaacctcc tattaatcaa attttaccaa ccttaactcc aattttaaga aacaagcata
     2401 gaaaagtcga agtaaacact atcaaatttg ttggacttat tggtaaatta gcacctacct
     2461 atgcacctcc taaagaatgg atgagaatat gttttgaact attagaactt ttgaaaagta
     2521 caaataaaga aataagaaga tcagcagctg ccacgtttgg attcattgct gaagcaattg
     2581 gacctcacga tgttcttgtt gcgttattga acaatttaaa ggttcaagaa cgtcagttac
     2641 gtgtatgcac ggctgttgct atcggtatcg ttgctaaagt gtgtggtcct tacaacgtgc
     2701 tgccagtaat catgaatgaa tacacaacac cagagactaa tgttcaaaac ggtgttctca
     2761 aggccatgtc attcatgttt gagtacattg gtaacatgtc taaggattac atatacttca
     2821 taacaccatt attagaagat gcactcaccg atagggattt agttcatcgt cagacagcgt
     2881 cgaacgtaat aacccactta gctttaaact gttcaggtac aggtcacgaa gatgccttca
     2941 ttcatttaat gaatcttcta atacccaata tttttgaaac ttcaccacat gctattatgc
     3001 gtattttaga aggtttggag gcactgagtc aagctcttgg ccctggacta ttcatgaact
     3061 acatatgggc aggattgttc catccagcga agaatgtaag gaaagcattt tggagggtgt
     3121 ataataacat gtacgtaatg taccaggacg ccatggtacc tttttacccc gttacaccag
     3181 acaacaatga agaatatata gaagaactgg atttagttct gtgaatatat atagagcata
     3241 taaataacca caacttgtaa gaatatttta cttgacacac ctatatcatc acgtcacgat
     3301 gccttcatag attatgatga aatataaaca gaacctgcta ttggaaaaaa aagggcagga
     3361 aaaaattttg tgaaagtgag taaaagaaag atgaagagcg agacatggat aataagaaat
     3421 agatgtcact aatggacaat tggaagactg atatggaaag ttacgatgaa ggaggagctc
     3481 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct
     3541 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3601 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3661 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3721 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3781 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3841 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3901 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3961 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4021 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4081 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4141 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4201 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4261 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4321 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4381 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4441 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4501 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4561 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4621 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4681 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4741 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4801 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4861 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4921 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4981 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     5041 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     5101 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5161 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5221 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5281 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5341 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5401 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5461 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5521 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5581 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5641 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgggtcct tttcatcacg
     5701 tgctataaaa ataattataa tttaaatttt ttaatataaa tatataaatt aaaaatagaa
     5761 agtaaaaaaa gaaattaaag aaaaaatagt ttttgttttc cgaagatgta aaagactcta
     5821 gggggatcgc caacaaatac taccttttat cttgctcttc ctgctctcag gtattaatgc
     5881 cgaattgttt catcttgtct gtgtagaaga ccacacacga aaatcctgtg attttacatt
     5941 ttacttatcg ttaatcgaat gtatatctat ttaatctgct tttcttgtct aataaatata
     6001 tatgtaaagt acgctttttg ttgaaatttt ttaaaccttt gtttattttt ttttcttcat
     6061 tccgtaactc ttctaccttc tttatttact ttctaaaatc caaatacaaa acataaaaat
     6121 aaataaacac agagtaaatt cccaaattat tccatcatta aaagatacga ggcgcgtgta
     6181 agttacaggc aagcgatccg tctaagaaac cattattatc atgacattaa cctataaaaa
     6241 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6301 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6361 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6421 atcagagcag attgtactga gagtgcacca tagatcacga cattactata tatataatat
     6481 aggaagcatt taatagaaca gcatcgtaat atatgtgtac tttgcagtta tgacgccaga
     6541 tggcagtagt ggaagatatt ctttattgaa aaatagcttg tcaccttacg tacaatcttg
     6601 atccggagct tttctttttt tgccgattaa gaattaattc ggtcgaaaaa agaaaaggag
     6661 agggccaaga gggagggcat tggtgactat tgagcacgtg agtatacgtg attaagcaca
     6721 caaaggcagc ttggagtatg tctgttatta atttcacagg tagttctggt ccattggtga
     6781 aagtttgcgg cttgcagagc acagaggccg cagaatgtgc tctagattcc gatgctgact
     6841 tgctgggtat tatatgtgtg cccaatagaa agagaacaat tgacccggtt attgcaagga
     6901 aaatttcaag tcttgtaaaa gcatataaaa atagttcagg cactccgaaa tacttggttg
     6961 gcgtgtttcg taatcaacct aaggaggatg ttttggctct ggtcaatgat tacggcattg
     7021 atatcgtcca actgcatgga gatgagtcgt ggcaagaata ccaagagttc ctcggtttgc
     7081 cagttattaa aagactcgta tttccaaaag actgcaacat actactcagt gcagcttcac
     7141 agaaacctca ttcgtttatt cccttgtttg attcagaagc aggtgggaca ggtgaacttt
     7201 tggattggaa ctcgatttct gactgggttg gaaggcaaga gagccccgaa agcttacatt
     7261 ttatgttagc tggtggactg acgccagaaa atgttggtga tgcgcttaga ttaaatggcg
     7321 ttattggtgt tgatgtaagc ggaggtgtgg agacaaatgg tgtaaaagac tctaacaaaa
     7381 tagcaaattt cgtcaaaaat gctaagaaat aggttattac tgagtagtat ttatttaagt
     7441 attgtttgtg cacttgccga tctatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     7501 aataccgcat caggaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg
     7561 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     7621 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     7681 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     7741 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     7801 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     7861 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     7921 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca
     7981 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     8041 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     8101 tcacgacgtt gtaaaacgac ggccagt
//
