LOCUS       sequence-213728-        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 R778A 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 R778A 3xFLAG
FEATURES             Location/Qualifiers
     source          1..8211
                     /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"
     CDS             3257..3322
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3594..3614)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3594..3612)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3633..3649)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3633..3649)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3646..3668)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3657..3673
                     /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(3681..3687,3688..3705,3706..3711))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3726..3747
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3864..3881)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4035..4623)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4115..4134)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4794..5585,5586..5654))
                     /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     5417..5436
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5655..5759)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5796..6299
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6340..6358
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6396..6418)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6518..6537
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6558..6838
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6839..7513
                     /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(7616..8071)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7753..7774)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7965..7984
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(8197..8211,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8211,1..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 ggcaatacag cgagtaggag gacttctatg cggattttaa 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 cagttagctg 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 tagttctggg
     3241 tggttctggt ggttctgatt acaaagacca cgatggtgac tacaaagacc atgatataga
     3301 ttacaaggat gatgatgata aatgaatata tatagagcat ataaataacc acaacttgta
     3361 agaatatttt acttgacaca cctatatcat cacgtcacga tgccttcata gattatgatg
     3421 aaatataaac agaacctgct attggaaaaa aaagggcagg aaaaaatttt gtgaaagtga
     3481 gtaaaagaaa gatgaagagc gagacatgga taataagaaa tagatgtcac taatggacaa
     3541 ttggaagact gatatggaaa gttacgatga aggaggagct ccagcttttg ttccctttag
     3601 tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     3661 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     3721 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     3781 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     3841 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     3901 cggcgagctg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     3961 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4021 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4081 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4141 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4201 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4261 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4321 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4381 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4441 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4501 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4561 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4621 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4681 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     4741 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     4801 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     4861 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     4921 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     4981 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5041 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5101 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5161 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5221 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5281 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5341 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5401 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5461 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5521 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5581 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5641 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     5701 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     5761 acatttcccc gaaaagtgcc acctgggtcc ttttcatcac gtgctataaa aataattata
     5821 atttaaattt tttaatataa atatataaat taaaaataga aagtaaaaaa agaaattaaa
     5881 gaaaaaatag tttttgtttt ccgaagatgt aaaagactct agggggatcg ccaacaaata
     5941 ctacctttta tcttgctctt cctgctctca ggtattaatg ccgaattgtt tcatcttgtc
     6001 tgtgtagaag accacacacg aaaatcctgt gattttacat tttacttatc gttaatcgaa
     6061 tgtatatcta tttaatctgc ttttcttgtc taataaatat atatgtaaag tacgcttttt
     6121 gttgaaattt tttaaacctt tgtttatttt tttttcttca ttccgtaact cttctacctt
     6181 ctttatttac tttctaaaat ccaaatacaa aacataaaaa taaataaaca cagagtaaat
     6241 tcccaaatta ttccatcatt aaaagatacg aggcgcgtgt aagttacagg caagcgatcc
     6301 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     6361 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     6421 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     6481 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     6541 agagtgcacc atagatcacg acattactat atatataata taggaagcat ttaatagaac
     6601 agcatcgtaa tatatgtgta ctttgcagtt atgacgccag atggcagtag tggaagatat
     6661 tctttattga aaaatagctt gtcaccttac gtacaatctt gatccggagc ttttcttttt
     6721 ttgccgatta agaattaatt cggtcgaaaa aagaaaagga gagggccaag agggagggca
     6781 ttggtgacta ttgagcacgt gagtatacgt gattaagcac acaaaggcag cttggagtat
     6841 gtctgttatt aatttcacag gtagttctgg tccattggtg aaagtttgcg gcttgcagag
     6901 cacagaggcc gcagaatgtg ctctagattc cgatgctgac ttgctgggta ttatatgtgt
     6961 gcccaataga aagagaacaa ttgacccggt tattgcaagg aaaatttcaa gtcttgtaaa
     7021 agcatataaa aatagttcag gcactccgaa atacttggtt ggcgtgtttc gtaatcaacc
     7081 taaggaggat gttttggctc tggtcaatga ttacggcatt gatatcgtcc aactgcatgg
     7141 agatgagtcg tggcaagaat accaagagtt cctcggtttg ccagttatta aaagactcgt
     7201 atttccaaaa gactgcaaca tactactcag tgcagcttca cagaaacctc attcgtttat
     7261 tcccttgttt gattcagaag caggtgggac aggtgaactt ttggattgga actcgatttc
     7321 tgactgggtt ggaaggcaag agagccccga aagcttacat tttatgttag ctggtggact
     7381 gacgccagaa aatgttggtg atgcgcttag attaaatggc gttattggtg ttgatgtaag
     7441 cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt tcgtcaaaaa
     7501 tgctaagaaa taggttatta ctgagtagta tttatttaag tattgtttgt gcacttgccg
     7561 atctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     7621 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     7681 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     7741 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     7801 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     7861 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     7921 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     7981 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     8041 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     8101 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     8161 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt t
//
