LOCUS       sequence-213557-        8128 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 K818A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8128)
  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 8128)
  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 K818A
FEATURES             Location/Qualifiers
     source          1..8128
                     /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(3511..3531)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3511..3529)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3550..3566)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3550..3566)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3563..3585)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3574..3590
                     /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(3598..3604,3605..3622,3623..3628))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3643..3664
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3781..3798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3952..4540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4032..4051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4711..5502,5503..5571))
                     /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     5334..5353
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5572..5676)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5713..6216
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6257..6275
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6313..6335)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6435..6454
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6475..6755
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6756..7430
                     /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(7533..7988)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7670..7691)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7882..7901
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(8114..8128,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8128,1..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 cagttacgtg tatgcacggc tgttgctatc ggtatcgttg ctaaagtgtg
     2701 tggtccttac aacgtgctgc cagtaatcat gaatgaatac acaacaccag agactaatgt
     2761 tcaaaacggt gttctcgctg 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 gaaaaaaaaa gggcaggaaa aaattttgtg aaagtgagta aaagaaagat gaagagcgag
     3421 acatggataa taagaaatag atgtcactaa tggacaattg gaagactgat atggaaagtt
     3481 acgatgaagg aggagctcca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     3541 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3601 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3661 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3721 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3781 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3841 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4141 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4381 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4501 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4561 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4621 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4681 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4741 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4801 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4861 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4921 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4981 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5041 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5101 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5161 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5221 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5281 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5341 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5401 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5461 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5521 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5581 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5641 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5701 tgggtccttt tcatcacgtg ctataaaaat aattataatt taaatttttt aatataaata
     5761 tataaattaa aaatagaaag taaaaaaaga aattaaagaa aaaatagttt ttgttttccg
     5821 aagatgtaaa agactctagg gggatcgcca acaaatacta ccttttatct tgctcttcct
     5881 gctctcaggt attaatgccg aattgtttca tcttgtctgt gtagaagacc acacacgaaa
     5941 atcctgtgat tttacatttt acttatcgtt aatcgaatgt atatctattt aatctgcttt
     6001 tcttgtctaa taaatatata tgtaaagtac gctttttgtt gaaatttttt aaacctttgt
     6061 ttattttttt ttcttcattc cgtaactctt ctaccttctt tatttacttt ctaaaatcca
     6121 aatacaaaac ataaaaataa ataaacacag agtaaattcc caaattattc catcattaaa
     6181 agatacgagg cgcgtgtaag ttacaggcaa gcgatccgtc taagaaacca ttattatcat
     6241 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     6301 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     6361 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     6421 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata gatcacgaca
     6481 ttactatata tataatatag gaagcattta atagaacagc atcgtaatat atgtgtactt
     6541 tgcagttatg acgccagatg gcagtagtgg aagatattct ttattgaaaa atagcttgtc
     6601 accttacgta caatcttgat ccggagcttt tctttttttg ccgattaaga attaattcgg
     6661 tcgaaaaaag aaaaggagag ggccaagagg gagggcattg gtgactattg agcacgtgag
     6721 tatacgtgat taagcacaca aaggcagctt ggagtatgtc tgttattaat ttcacaggta
     6781 gttctggtcc attggtgaaa gtttgcggct tgcagagcac agaggccgca gaatgtgctc
     6841 tagattccga tgctgacttg ctgggtatta tatgtgtgcc caatagaaag agaacaattg
     6901 acccggttat tgcaaggaaa atttcaagtc ttgtaaaagc atataaaaat agttcaggca
     6961 ctccgaaata cttggttggc gtgtttcgta atcaacctaa ggaggatgtt ttggctctgg
     7021 tcaatgatta cggcattgat atcgtccaac tgcatggaga tgagtcgtgg caagaatacc
     7081 aagagttcct cggtttgcca gttattaaaa gactcgtatt tccaaaagac tgcaacatac
     7141 tactcagtgc agcttcacag aaacctcatt cgtttattcc cttgtttgat tcagaagcag
     7201 gtgggacagg tgaacttttg gattggaact cgatttctga ctgggttgga aggcaagaga
     7261 gccccgaaag cttacatttt atgttagctg gtggactgac gccagaaaat gttggtgatg
     7321 cgcttagatt aaatggcgtt attggtgttg atgtaagcgg aggtgtggag acaaatggtg
     7381 taaaagactc taacaaaata gcaaatttcg tcaaaaatgc taagaaatag gttattactg
     7441 agtagtattt atttaagtat tgtttgtgca cttgccgatc tatgcggtgt gaaataccgc
     7501 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
     7561 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     7621 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     7681 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     7741 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     7801 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     7861 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     7921 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     7981 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     8041 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     8101 acgccagggt tttcccagtc acgacgtt
//
