LOCUS       sequence-213729-        8211 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Shuffle plasmid containing the HSH155 gene and surrounding regions. 
            It bears a TRP marker for shuffle into a HSH155 deletion strain. It 
            has 3xFLAG at the C-terminus. Contains R775A mutation. Lethal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8211)
  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 8211)
  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 R775A 3xFLAG
FEATURES             Location/Qualifiers
     source          1..8211
                     /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"
     CDS             3240..3305
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3577..3597)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3577..3595)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3616..3632)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3616..3632)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3629..3651)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3640..3656
                     /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(3664..3670,3671..3688,3689..3694))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3709..3730
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3847..3864)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4018..4606)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4098..4117)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4777..5568,5569..5637))
                     /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     5400..5419
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5638..5742)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5779..6282
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6323..6341
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6379..6401)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6501..6520
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6541..6821
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6822..7496
                     /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(7599..8054)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7736..7757)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7948..7967
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8180..8202
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8194..8211
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8195..8211
                     /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 tcagcaaatg ccacgtttgg attcattgct gaagcaattg
     2581 gacctcacga tgttcttgtt gcgttattga acaatttaaa ggttcaagaa gctcagttac
     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 gggtggttct ggtggttctg
     3241 attacaaaga ccacgatggt gactacaaag accatgatat agattacaag gatgatgatg
     3301 ataaatgaat atatatagag catataaata accacaactt gtaagaatat tttacttgac
     3361 acacctatat catcacgtca cgatgccttc atagattatg atgaaatata aacagaacct
     3421 gctattggaa aaaaaagggc aggaaaaaat tttgtgaaag tgagtaaaag aaagatgaag
     3481 agcgagacat ggataataag aaatagatgt cactaatgga caattggaag actgatatgg
     3541 aaagttacga tgaaggagga gctccagctt ttgttccctt tagtgagggt taattgcgcg
     3601 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3661 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3721 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3781 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3841 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     3901 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     3961 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4021 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4081 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4141 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4201 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4261 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4321 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4381 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4441 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4501 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4561 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4621 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4681 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4741 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4801 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4861 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     4921 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     4981 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5041 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5101 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5161 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5221 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5281 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5341 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5401 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5461 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5521 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5581 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5641 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5701 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5761 gccacctggg tccttttcat cacgtgctat aaaaataatt ataatttaaa ttttttaata
     5821 taaatatata aattaaaaat agaaagtaaa aaaagaaatt aaagaaaaaa tagtttttgt
     5881 tttccgaaga tgtaaaagac tctaggggga tcgccaacaa atactacctt ttatcttgct
     5941 cttcctgctc tcaggtatta atgccgaatt gtttcatctt gtctgtgtag aagaccacac
     6001 acgaaaatcc tgtgatttta cattttactt atcgttaatc gaatgtatat ctatttaatc
     6061 tgcttttctt gtctaataaa tatatatgta aagtacgctt tttgttgaaa ttttttaaac
     6121 ctttgtttat ttttttttct tcattccgta actcttctac cttctttatt tactttctaa
     6181 aatccaaata caaaacataa aaataaataa acacagagta aattcccaaa ttattccatc
     6241 attaaaagat acgaggcgcg tgtaagttac aggcaagcga tccgtctaag aaaccattat
     6301 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6361 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6421 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6481 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatagatc
     6541 acgacattac tatatatata atataggaag catttaatag aacagcatcg taatatatgt
     6601 gtactttgca gttatgacgc cagatggcag tagtggaaga tattctttat tgaaaaatag
     6661 cttgtcacct tacgtacaat cttgatccgg agcttttctt tttttgccga ttaagaatta
     6721 attcggtcga aaaaagaaaa ggagagggcc aagagggagg gcattggtga ctattgagca
     6781 cgtgagtata cgtgattaag cacacaaagg cagcttggag tatgtctgtt attaatttca
     6841 caggtagttc tggtccattg gtgaaagttt gcggcttgca gagcacagag gccgcagaat
     6901 gtgctctaga ttccgatgct gacttgctgg gtattatatg tgtgcccaat agaaagagaa
     6961 caattgaccc ggttattgca aggaaaattt caagtcttgt aaaagcatat aaaaatagtt
     7021 caggcactcc gaaatacttg gttggcgtgt ttcgtaatca acctaaggag gatgttttgg
     7081 ctctggtcaa tgattacggc attgatatcg tccaactgca tggagatgag tcgtggcaag
     7141 aataccaaga gttcctcggt ttgccagtta ttaaaagact cgtatttcca aaagactgca
     7201 acatactact cagtgcagct tcacagaaac ctcattcgtt tattcccttg tttgattcag
     7261 aagcaggtgg gacaggtgaa cttttggatt ggaactcgat ttctgactgg gttggaaggc
     7321 aagagagccc cgaaagctta cattttatgt tagctggtgg actgacgcca gaaaatgttg
     7381 gtgatgcgct tagattaaat ggcgttattg gtgttgatgt aagcggaggt gtggagacaa
     7441 atggtgtaaa agactctaac aaaatagcaa atttcgtcaa aaatgctaag aaataggtta
     7501 ttactgagta gtatttattt aagtattgtt tgtgcacttg ccgatctatg cggtgtgaaa
     7561 taccgcacag atgcgtaagg agaaaatacc gcatcaggaa attgtaagcg ttaatatttt
     7621 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     7681 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     7741 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     7801 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     7861 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     7921 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     7981 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     8041 gctacagggc gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     8101 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
     8161 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag t
//
