LOCUS       sequence-213582-        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 R744A 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 R744A
FEATURES             Location/Qualifiers
     source          1..8127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3352..3372)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3352..3370)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3391..3407)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3391..3407)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3404..3426)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3415..3431
                     /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(3439..3445,3446..3463,3464..3469))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3484..3505
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3622..3639)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3793..4381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3873..3892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4552..5343,5344..5412))
                     /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     5175..5194
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5413..5517)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5554..6057
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6098..6116
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6154..6176)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6276..6295
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6316..6596
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             6597..7271
                     /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(7374..7829)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7511..7532)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7723..7742
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7955..7977
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7969..7986
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7970..7986
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7996..8015
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7996..8014
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gctttttaga cgtgattgct gttatttggc acgttgttag tatttaagct tattcgggtt
       61 tgcattgata tttgcccgag cagaaatctc cttctcgatg agttagaact gaaacatctc
      121 ttcaattact tatacataat aaacatagaa agccaggacc tgtcatcatg agtcatccga
      181 ttcaatttgt aaacgctaat aatagcgata agagccacca attaggcggt caatattcta
      241 ttcctcagga tttgagggaa aacctgcaga aagaagctgc acgtattggt gagaacgaaa
      301 aagatgtgct acaggaaaaa atggaaacta ggacagtgca aaatcgagag gattcatacc
      361 acaaaagaag gtttgatatg aagtttgagc cggattccga tacacaaact gttacgtctt
      421 cagaaaatac ccaagacgct gtcgtgccaa ggaaaagaaa atctcgttgg gatgtaaaag
      481 ggtacgagcc acctgatgaa tcttcaacag cggtcaaaga aaactctgat agtgctttgg
      541 ttaatgtaga aggaatccat gatttgatgt tttttaaacc ttctgaccat aaatatttcg
      601 ccgacgttat atccaaaaaa cccatagatg agttaaataa ggatgaaaag aaagaaagaa
      661 ccctgtcgat gttacttctg aaaataaaaa atggcaatac agcgagtagg aggacttcta
      721 tgcggatttt aaccgataaa gctgttacat ttggtccaga aatgatattt aaccgcttat
      781 tgcccatatt actagacaga agtttggaag accaggaaag acatctaatg atcaagacca
      841 tagatcgcgt cctttaccag ctaggagatt taacaaaacc ttacgttcac aaaatacttg
      901 tagtagctgc cccattgcta atagatgaag atccaatggt tcgttcaaca ggccaggaga
      961 taatcaccaa tctgtctaca gtcgctggcc taaaaaccat tctaacagta atgagacctg
     1021 atattgagaa tgaggatgag tatgttagaa atgtaacttc aagagcggcg gctgtggtgg
     1081 caaaagccct tggtgttaat caattgcttc ctttcatcaa tgcagcatgc cactcacgga
     1141 agtcttggaa agcaaggcat actggcataa aaattgtgca gcaaataggt atacttttgg
     1201 gaataggtgt cttgaatcat cttactgggc tgatgagttg tatcaaagac tgcctaatgg
     1261 atgatcacgt tccggtgcga attgttacag cacatacttt atcaacacta gcagaaaatt
     1321 cttatccata tggtatcgaa gttttcaacg ttgtactgga gccattatgg aagggtataa
     1381 gaagtcatcg tggtaaagtg ctatcttctt ttttaaaagc tgttggatct atgatcccat
     1441 tgatggaccc agagtatgcc ggttattata ccacagaagc aatgagaata attagacgtg
     1501 aattcgactc tcccgatgat gaaatgaaaa agaccattct tttagtatta caaaaatgta
     1561 gcgctgtaga atccataacg ccaaaatttc tgagagaaga aatcgcgcca gaattctttc
     1621 aaaaattttg ggtcagacga gtggcattgg atagaccatt aaacaaagtt gttacatata
     1681 ctactgttac attagcaaag aaattaggtt gttcttatac cattgataaa ttattaacac
     1741 cattgagaga tgaagctgag ccgttcagaa caatggctgt ccatgctgtt actagaacag
     1801 taaacttatt agggacagca gatttagatg agagattaga aacgaggctt attgatgcgc
     1861 tcttgatagc atttcaagaa caaacaaata gtgattctat aatctttaaa gggtttgggg
     1921 cagtgacggt gtcacttgac atccgtatga aaccgttttt agcacccatt gtgagtacga
     1981 ttttaaatca tttaaagcat aaaactccct tggttcgtca acatgcagca gatctatgtg
     2041 ccatactaat accagtcatc aagaactgtc acgaattcga aatgttgaat aaattgaaca
     2101 ttatattgta tgaatcctta ggcgaagtat atccagaggt attaggttct ataatcaacg
     2161 ctatgtactg tattactagc gtaatggatt tggataaact acaacctcct attaatcaaa
     2221 ttttaccaac cttaactcca attttaagaa acaagcatag aaaagtcgaa gtaaacacta
     2281 tcaaatttgt tggacttatt ggtaaattag cacctaccta tgcacctcct aaagaatgga
     2341 tgagaatatg ttttgaacta ttagaacttt tgaaaagtac aaataaagaa ataagagctt
     2401 cagcaaatgc cacgtttgga ttcattgctg aagcaattgg acctcacgat gttcttgttg
     2461 cgttattgaa caatttaaag gttcaagaac gtcagttacg tgtatgcacg gctgttgcta
     2521 tcggtatcgt tgctaaagtg tgtggtcctt acaacgtgct gccagtaatc atgaatgaat
     2581 acacaacacc agagactaat gttcaaaacg gtgttctcaa ggccatgtca ttcatgtttg
     2641 agtacattgg taacatgtct aaggattaca tatacttcat aacaccatta ttagaagatg
     2701 cactcaccga tagggattta gttcatcgtc agacagcgtc gaacgtaata acccacttag
     2761 ctttaaactg ttcaggtaca ggtcacgaag atgccttcat tcatttaatg aatcttctaa
     2821 tacccaatat ttttgaaact tcaccacatg ctattatgcg tattttagaa ggtttggagg
     2881 cactgagtca agctcttggc cctggactat tcatgaacta catatgggca ggattgttcc
     2941 atccagcgaa gaatgtaagg aaagcatttt ggagggtgta taataacatg tacgtaatgt
     3001 accaggacgc catggtacct ttttaccccg ttacaccaga caacaatgaa gaatatatag
     3061 aagaactgga tttagttctg tgaatatata tagagcatat aaataaccac aacttgtaag
     3121 aatattttac ttgacacacc tatatcatca cgtcacgatg ccttcataga ttatgatgaa
     3181 atataaacag aacctgctat tggaaaaaaa agggcaggaa aaaattttgt gaaagtgagt
     3241 aaaagaaaga tgaagagcga gacatggata ataagaaata gatgtcacta atggacaatt
     3301 ggaagactga tatggaaagt tacgatgaag gaggagctcc agcttttgtt ccctttagtg
     3361 agggttaatt gcgcgcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     3421 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     3481 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     3541 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     3601 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3661 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3721 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3781 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3841 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3901 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3961 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4021 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4081 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4141 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4201 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     4261 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4321 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4381 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4441 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4501 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4561 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4621 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4681 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4741 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4801 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4861 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4921 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4981 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     5041 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     5101 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     5161 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     5221 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     5281 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     5341 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     5401 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     5461 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5521 atttccccga aaagtgccac ctgggtcctt ttcatcacgt gctataaaaa taattataat
     5581 ttaaattttt taatataaat atataaatta aaaatagaaa gtaaaaaaag aaattaaaga
     5641 aaaaatagtt tttgttttcc gaagatgtaa aagactctag ggggatcgcc aacaaatact
     5701 accttttatc ttgctcttcc tgctctcagg tattaatgcc gaattgtttc atcttgtctg
     5761 tgtagaagac cacacacgaa aatcctgtga ttttacattt tacttatcgt taatcgaatg
     5821 tatatctatt taatctgctt ttcttgtcta ataaatatat atgtaaagta cgctttttgt
     5881 tgaaattttt taaacctttg tttatttttt tttcttcatt ccgtaactct tctaccttct
     5941 ttatttactt tctaaaatcc aaatacaaaa cataaaaata aataaacaca gagtaaattc
     6001 ccaaattatt ccatcattaa aagatacgag gcgcgtgtaa gttacaggca agcgatccgt
     6061 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     6121 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     6181 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     6241 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     6301 agtgcaccat agatcacgac attactatat atataatata ggaagcattt aatagaacag
     6361 catcgtaata tatgtgtact ttgcagttat gacgccagat ggcagtagtg gaagatattc
     6421 tttattgaaa aatagcttgt caccttacgt acaatcttga tccggagctt ttcttttttt
     6481 gccgattaag aattaattcg gtcgaaaaaa gaaaaggaga gggccaagag ggagggcatt
     6541 ggtgactatt gagcacgtga gtatacgtga ttaagcacac aaaggcagct tggagtatgt
     6601 ctgttattaa tttcacaggt agttctggtc cattggtgaa agtttgcggc ttgcagagca
     6661 cagaggccgc agaatgtgct ctagattccg atgctgactt gctgggtatt atatgtgtgc
     6721 ccaatagaaa gagaacaatt gacccggtta ttgcaaggaa aatttcaagt cttgtaaaag
     6781 catataaaaa tagttcaggc actccgaaat acttggttgg cgtgtttcgt aatcaaccta
     6841 aggaggatgt tttggctctg gtcaatgatt acggcattga tatcgtccaa ctgcatggag
     6901 atgagtcgtg gcaagaatac caagagttcc tcggtttgcc agttattaaa agactcgtat
     6961 ttccaaaaga ctgcaacata ctactcagtg cagcttcaca gaaacctcat tcgtttattc
     7021 ccttgtttga ttcagaagca ggtgggacag gtgaactttt ggattggaac tcgatttctg
     7081 actgggttgg aaggcaagag agccccgaaa gcttacattt tatgttagct ggtggactga
     7141 cgccagaaaa tgttggtgat gcgcttagat taaatggcgt tattggtgtt gatgtaagcg
     7201 gaggtgtgga gacaaatggt gtaaaagact ctaacaaaat agcaaatttc gtcaaaaatg
     7261 ctaagaaata ggttattact gagtagtatt tatttaagta ttgtttgtgc acttgccgat
     7321 ctatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     7381 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     7441 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     7501 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     7561 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     7621 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     7681 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     7741 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     7801 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     7861 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     7921 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     7981 gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg gcccccccct
     8041 cgagcatgta ctttccgcct aactaccatc acctgtcttt atttattccg ctgacagata
     8101 tgcacaacca attatcgtct cttgaaa
//
