LOCUS       sequence-213574-        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 T178A R181A R182A R186A 
            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 T178A R181A R182A R186A (N4A)
FEATURES             Location/Qualifiers
     source          1..8127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     27..46
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        27..45
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3510..3530)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3510..3528)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3549..3565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3549..3565)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3562..3584)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3573..3589
                     /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(3597..3603,3604..3621,3622..3627))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3642..3663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3780..3797)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3951..4539)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4031..4050)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4710..5501,5502..5570))
                     /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     5333..5352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5571..5675)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5712..6215
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6256..6274
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6312..6334)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6434..6453
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6474..6754
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6755..7429
                     /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(7532..7987)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7669..7690)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7881..7900
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(8113..8127,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8127,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtaccg
       61 ggcccccccc tcgagcatgt actttccgcc taactaccat cacctgtctt tatttattcc
      121 gctgacagat atgcacaacc aattatcgtc tcttgaaagc tttttagacg tgattgctgt
      181 tatttggcac gttgttagta tttaagctta ttcgggtttg cattgatatt tgcccgagca
      241 gaaatctcct tctcgatgag ttagaactga aacatctctt caattactta tacataataa
      301 acatagaaag ccaggacctg tcatcatgag tcatccgatt caatttgtaa acgctaataa
      361 tagcgataag agccaccaat taggcggtca atattctatt cctcaggatt tgagggaaaa
      421 cctgcagaaa gaagctgcac gtattggtga gaacgaaaaa gatgtgctac aggaaaaaat
      481 ggaaactagg acagtgcaaa atcgagagga ttcataccac aaaagaaggt ttgatatgaa
      541 gtttgagccg gattccgata cacaaactgt tacgtcttca gaaaataccc aagacgctgt
      601 cgtgccaagg aaaagaaaat ctcgttggga tgtaaaaggg tacgagccac ctgatgaatc
      661 ttcaacagcg gtcaaagaaa actctgatag tgctttggtt aatgtagaag gaatccatga
      721 tttgatgttt tttaaacctt ctgaccataa atatttcgcc gacgttatat ccaaaaaacc
      781 catagatgag ttaaataagg atgaaaagaa agaaagaacc ctgtcgatgt tacttctgaa
      841 aataaaaaat ggcaatgctg cgagtgctgc tacttctatg gctattttaa ccgataaagc
      901 tgttacattt ggtccagaaa tgatatttaa ccgcttattg cccatattac tagacagaag
      961 tttggaagac caggaaagac atctaatgat caagaccata gatcgcgtcc tttaccagct
     1021 aggagattta acaaaacctt acgttcacaa aatacttgta gtagctgccc cattgctaat
     1081 agatgaagat ccaatggttc gttcaacagg ccaggagata atcaccaatc tgtctacagt
     1141 cgctggccta aaaaccattc taacagtaat gagacctgat attgagaatg aggatgagta
     1201 tgttagaaat gtaacttcaa gagcggcggc tgtggtggca aaagcccttg gtgttaatca
     1261 attgcttcct ttcatcaatg cagcatgcca ctcacggaag tcttggaaag caaggcatac
     1321 tggcataaaa attgtgcagc aaataggtat acttttggga ataggtgtct tgaatcatct
     1381 tactgggctg atgagttgta tcaaagactg cctaatggat gatcacgttc cggtgcgaat
     1441 tgttacagca catactttat caacactagc agaaaattct tatccatatg gtatcgaagt
     1501 tttcaacgtt gtactggagc cattatggaa gggtataaga agtcatcgtg gtaaagtgct
     1561 atcttctttt ttaaaagctg ttggatctat gatcccattg atggacccag agtatgccgg
     1621 ttattatacc acagaagcaa tgagaataat tagacgtgaa ttcgactctc ccgatgatga
     1681 aatgaaaaag accattcttt tagtattaca aaaatgtagc gctgtagaat ccataacgcc
     1741 aaaatttctg agagaagaaa tcgcgccaga attctttcaa aaattttggg tcagacgagt
     1801 ggcattggat agaccattaa acaaagttgt tacatatact actgttacat tagcaaagaa
     1861 attaggttgt tcttatacca ttgataaatt attaacacca ttgagagatg aagctgagcc
     1921 gttcagaaca atggctgtcc atgctgttac tagaacagta aacttattag ggacagcaga
     1981 tttagatgag agattagaaa cgaggcttat tgatgcgctc ttgatagcat ttcaagaaca
     2041 aacaaatagt gattctataa tctttaaagg gtttggggca gtgacggtgt cacttgacat
     2101 ccgtatgaaa ccgtttttag cacccattgt gagtacgatt ttaaatcatt taaagcataa
     2161 aactcccttg gttcgtcaac atgcagcaga tctatgtgcc atactaatac cagtcatcaa
     2221 gaactgtcac gaattcgaaa tgttgaataa attgaacatt atattgtatg aatccttagg
     2281 cgaagtatat ccagaggtat taggttctat aatcaacgct atgtactgta ttactagcgt
     2341 aatggatttg gataaactac aacctcctat taatcaaatt ttaccaacct taactccaat
     2401 tttaagaaac aagcatagaa aagtcgaagt aaacactatc aaatttgttg gacttattgg
     2461 taaattagca cctacctatg cacctcctaa agaatggatg agaatatgtt ttgaactatt
     2521 agaacttttg aaaagtacaa ataaagaaat aagaagatca gcaaatgcca cgtttggatt
     2581 cattgctgaa gcaattggac ctcacgatgt tcttgttgcg ttattgaaca atttaaaggt
     2641 tcaagaacgt cagttacgtg tatgcacggc tgttgctatc ggtatcgttg ctaaagtgtg
     2701 tggtccttac aacgtgctgc cagtaatcat gaatgaatac acaacaccag agactaatgt
     2761 tcaaaacggt gttctcaagg ccatgtcatt catgtttgag tacattggta acatgtctaa
     2821 ggattacata tacttcataa caccattatt agaagatgca ctcaccgata gggatttagt
     2881 tcatcgtcag acagcgtcga acgtaataac ccacttagct ttaaactgtt caggtacagg
     2941 tcacgaagat gccttcattc atttaatgaa tcttctaata cccaatattt ttgaaacttc
     3001 accacatgct attatgcgta ttttagaagg tttggaggca ctgagtcaag ctcttggccc
     3061 tggactattc atgaactaca tatgggcagg attgttccat ccagcgaaga atgtaaggaa
     3121 agcattttgg agggtgtata ataacatgta cgtaatgtac caggacgcca tggtaccttt
     3181 ttaccccgtt acaccagaca acaatgaaga atatatagaa gaactggatt tagttctgtg
     3241 aatatatata gagcatataa ataaccacaa cttgtaagaa tattttactt gacacaccta
     3301 tatcatcacg tcacgatgcc ttcatagatt atgatgaaat ataaacagaa cctgctattg
     3361 gaaaaaaaag ggcaggaaaa aattttgtga aagtgagtaa aagaaagatg aagagcgaga
     3421 catggataat aagaaataga tgtcactaat ggacaattgg aagactgata tggaaagtta
     3481 cgatgaagga ggagctccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     3541 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3601 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3661 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3721 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3781 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3841 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3901 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3961 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4021 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4081 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4141 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4201 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4261 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4321 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4381 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4441 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4501 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4561 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4621 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4681 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4741 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4801 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     4861 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4921 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4981 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5041 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5101 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5161 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5221 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5281 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5341 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5401 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5461 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5521 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5581 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     5641 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     5701 gggtcctttt catcacgtgc tataaaaata attataattt aaatttttta atataaatat
     5761 ataaattaaa aatagaaagt aaaaaaagaa attaaagaaa aaatagtttt tgttttccga
     5821 agatgtaaaa gactctaggg ggatcgccaa caaatactac cttttatctt gctcttcctg
     5881 ctctcaggta ttaatgccga attgtttcat cttgtctgtg tagaagacca cacacgaaaa
     5941 tcctgtgatt ttacatttta cttatcgtta atcgaatgta tatctattta atctgctttt
     6001 cttgtctaat aaatatatat gtaaagtacg ctttttgttg aaatttttta aacctttgtt
     6061 tatttttttt tcttcattcc gtaactcttc taccttcttt atttactttc taaaatccaa
     6121 atacaaaaca taaaaataaa taaacacaga gtaaattccc aaattattcc atcattaaaa
     6181 gatacgaggc gcgtgtaagt tacaggcaag cgatccgtct aagaaaccat tattatcatg
     6241 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     6301 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     6361 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     6421 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatag atcacgacat
     6481 tactatatat ataatatagg aagcatttaa tagaacagca tcgtaatata tgtgtacttt
     6541 gcagttatga cgccagatgg cagtagtgga agatattctt tattgaaaaa tagcttgtca
     6601 ccttacgtac aatcttgatc cggagctttt ctttttttgc cgattaagaa ttaattcggt
     6661 cgaaaaaaga aaaggagagg gccaagaggg agggcattgg tgactattga gcacgtgagt
     6721 atacgtgatt aagcacacaa aggcagcttg gagtatgtct gttattaatt tcacaggtag
     6781 ttctggtcca ttggtgaaag tttgcggctt gcagagcaca gaggccgcag aatgtgctct
     6841 agattccgat gctgacttgc tgggtattat atgtgtgccc aatagaaaga gaacaattga
     6901 cccggttatt gcaaggaaaa tttcaagtct tgtaaaagca tataaaaata gttcaggcac
     6961 tccgaaatac ttggttggcg tgtttcgtaa tcaacctaag gaggatgttt tggctctggt
     7021 caatgattac ggcattgata tcgtccaact gcatggagat gagtcgtggc aagaatacca
     7081 agagttcctc ggtttgccag ttattaaaag actcgtattt ccaaaagact gcaacatact
     7141 actcagtgca gcttcacaga aacctcattc gtttattccc ttgtttgatt cagaagcagg
     7201 tgggacaggt gaacttttgg attggaactc gatttctgac tgggttggaa ggcaagagag
     7261 ccccgaaagc ttacatttta tgttagctgg tggactgacg ccagaaaatg ttggtgatgc
     7321 gcttagatta aatggcgtta ttggtgttga tgtaagcgga ggtgtggaga caaatggtgt
     7381 aaaagactct aacaaaatag caaatttcgt caaaaatgct aagaaatagg ttattactga
     7441 gtagtattta tttaagtatt gtttgtgcac ttgccgatct atgcggtgtg aaataccgca
     7501 cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa
     7561 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     7621 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     7681 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     7741 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     7801 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     7861 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     7921 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     7981 ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     8041 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     8101 cgccagggtt ttcccagtca cgacgtt
//
