LOCUS       sequence-189170-        8221 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid used to maintain viability after deletion of genomic copy of
            HSH155..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8221)
  AUTHORS   Carrocci TJ, Zoerner DM, Paulson JC, Hoskins AA
  TITLE     SF3b1 mutations associated with myelodysplastic syndromes alter the 
            fidelity of branchsite selection in yeast.
  JOURNAL   Nucleic Acids Res. 2017 Jan 6. pii: gkw1349. doi: 
            10.1093/nar/gkw1349.
  PUBMED    28062854
REFERENCE   2  (bases 1 to 8221)
  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: "Nucleic 
            Acids Res. 2017 Jan 6. pii: gkw1349. doi: 10.1093/nar/gkw1349."
COMMENT     Sequence Label: pRS416 Hsh155 WT
FEATURES             Location/Qualifiers
     source          1..8221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        194..409
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             410..1213
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1344..1799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1481..1502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1693..1712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1925..1947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1939..1956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1940..1956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5448..5468)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5448..5466)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5487..5503)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5487..5503)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5500..5522)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5511..5527
                     /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(5535..5541,5542..5559,5560..5565))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5580..5601
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5718..5735)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5889..6477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5969..5988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6648..7439,7440..7508))
                     /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     7271..7290
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7509..7613)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    7650..8153
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     8194..8212
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1921 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     1981 tagggcgaat tgggtaccgg gccccccctc gagcatgtac tttccgccta actaccatca
     2041 cctgtcttta tttattccgc tgacagatat gcacaaccaa ttatcgtctc ttgaaagctt
     2101 tttagacgtg attgctgtta tttggcacgt tgttagtatt taagcttatt cgggtttgca
     2161 ttgatatttg cccgagcaga aatctccttc tcgatgagtt agaactgaaa catctcttca
     2221 attacttata cataataaac atagaaagcc aggacctgtc atcatgagtc atccgattca
     2281 atttgtaaac gctaataata gcgataagag ccaccaatta ggcggtcaat attctattcc
     2341 tcaggatttg agggaaaacc tgcagaaaga agctgcacgt attggtgaga acgaaaaaga
     2401 tgtgctacag gaaaaaatgg aaactaggac agtgcaaaat cgagaggatt cataccacaa
     2461 aagaaggttt gatatgaagt ttgagccgga ttccgataca caaactgtta cgtcttcaga
     2521 aaatacccaa gacgctgtcg tgccaaggaa aagaaaatct cgttgggatg taaaagggta
     2581 cgagccacct gatgaatctt caacagcggt caaagaaaac tctgatagtg ctttggttaa
     2641 tgtagaagga atccatgatt tgatgttttt taaaccttct gaccataaat atttcgccga
     2701 cgttatatcc aaaaaaccca tagatgagtt aaataaggat gaaaagaaag aaagaaccct
     2761 gtcgatgtta cttctgaaaa taaaaaatgg caatacagcg agtaggagga cttctatgcg
     2821 gattttaacc gataaagctg ttacatttgg tccagaaatg atatttaacc gcttattgcc
     2881 catattacta gacagaagtt tggaagacca ggaaagacat ctaatgatca agaccataga
     2941 tcgcgtcctt taccagctag gagatttaac aaaaccttac gttcacaaaa tacttgtagt
     3001 agctgcccca ttgctaatag atgaagatcc aatggttcgt tcaacaggcc aggagataat
     3061 caccaatctg tctacagtcg ctggcctaaa aaccattcta acagtaatga gacctgatat
     3121 tgagaatgag gatgagtatg ttagaaatgt aacttcaaga gcggcggctg tggtggcaaa
     3181 agcccttggt gttaatcaat tgcttccttt catcaatgca gcatgccact cacggaagtc
     3241 ttggaaagca aggcatactg gcataaaaat tgtgcagcaa ataggtatac ttttgggaat
     3301 aggtgtcttg aatcatctta ctgggctgat gagttgtatc aaagactgcc taatggatga
     3361 tcacgttccg gtgcgaattg ttacagcaca tactttatca acactagcag aaaattctta
     3421 tccatatggt atcgaagttt tcaacgttgt actggagcca ttatggaagg gtataagaag
     3481 tcatcgtggt aaagtgctat cttctttttt aaaagctgtt ggatctatga tcccattgat
     3541 ggacccagag tatgccggtt attataccac agaagcaatg agaataatta gacgtgaatt
     3601 cgactctccc gatgatgaaa tgaaaaagac cattctttta gtattacaaa aatgtagcgc
     3661 tgtagaatcc ataacgccaa aatttctgag agaagaaatc gcgccagaat tctttcaaaa
     3721 attttgggtc agacgagtgg cattggatag accattaaac aaagttgtta catatactac
     3781 tgttacatta gcaaagaaat taggttgttc ttataccatt gataaattat taacaccatt
     3841 gagagatgaa gctgagccgt tcagaacaat ggctgtccat gctgttacta gaacagtaaa
     3901 cttattaggg acagcagatt tagatgagag attagaaacg aggcttattg atgcgctctt
     3961 gatagcattt caagaacaaa caaatagtga ttctataatc tttaaagggt ttggggcagt
     4021 gacggtgtca cttgacatcc gtatgaaacc gtttttagca cccattgtga gtacgatttt
     4081 aaatcattta aagcataaaa ctcccttggt tcgtcaacat gcagcagatc tatgtgccat
     4141 actaatacca gtcatcaaga actgtcacga attcgaaatg ttgaataaat tgaacattat
     4201 attgtatgaa tccttaggcg aagtatatcc agaggtatta ggttctataa tcaacgctat
     4261 gtactgtatt actagcgtaa tggatttgga taaactacaa cctcctatta atcaaatttt
     4321 accaacctta actccaattt taagaaacaa gcatagaaaa gtcgaagtaa acactatcaa
     4381 atttgttgga cttattggta aattagcacc tacctatgca cctcctaaag aatggatgag
     4441 aatatgtttt gaactattag aacttttgaa aagtacaaat aaagaaataa gaagatcagc
     4501 aaatgccacg tttggattca ttgctgaagc aattggacct cacgatgttc ttgttgcgtt
     4561 attgaacaat ttaaaggttc aagaacgtca gttacgtgta tgcacggctg ttgctatcgg
     4621 tatcgttgct aaagtgtgtg gtccttacaa cgtgctgcca gtaatcatga atgaatacac
     4681 aacaccagag actaatgttc aaaacggtgt tctcaaggcc atgtcattca tgtttgagta
     4741 cattggtaac atgtctaagg attacatata cttcataaca ccattattag aagatgcact
     4801 caccgatagg gatttagttc atcgtcagac agcgtcgaac gtaataaccc acttagcttt
     4861 aaactgttca ggtacaggtc acgaagatgc cttcattcat ttaatgaatc ttctaatacc
     4921 caatattttt gaaacttcac cacatgctat tatgcgtatt ttagaaggtt tggaggcact
     4981 gagtcaagct cttggccctg gactattcat gaactacata tgggcaggat tgttccatcc
     5041 agcgaagaat gtaaggaaag cattttggag ggtgtataat aacatgtacg taatgtacca
     5101 ggacgccatg gtaccttttt accccgttac accagacaac aatgaagaat atatagaaga
     5161 actggattta gttctgtgaa tatatataga gcatataaat aaccacaact tgtaagaata
     5221 ttttacttga cacacctata tcatcacgtc acgatgcctt catagattat gatgaaatat
     5281 aaacagaacc tgctattgga aaaaaaaggg caggaaaaaa ttttgtgaaa gtgagtaaaa
     5341 gaaagatgaa gagcgagaca tggataataa gaaatagatg tcactaatgg acaattggaa
     5401 gactgatatg gaaagttacg atgaaggagg agctccagct tttgttccct ttagtgaggg
     5461 ttaattgcgc gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     5521 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     5581 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     5641 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5701 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5761 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5821 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5881 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5941 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6001 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6061 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6121 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6181 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6241 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6301 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6361 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6421 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6481 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6541 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6601 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6661 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6721 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6781 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6841 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6901 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6961 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7021 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7081 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7141 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7201 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7261 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     7321 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     7381 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     7441 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     7501 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7561 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7621 ccccgaaaag tgccacctgg gtccttttca tcacgtgcta taaaaataat tataatttaa
     7681 attttttaat ataaatatat aaattaaaaa tagaaagtaa aaaaagaaat taaagaaaaa
     7741 atagtttttg ttttccgaag atgtaaaaga ctctaggggg atcgccaaca aatactacct
     7801 tttatcttgc tcttcctgct ctcaggtatt aatgccgaat tgtttcatct tgtctgtgta
     7861 gaagaccaca cacgaaaatc ctgtgatttt acattttact tatcgttaat cgaatgtata
     7921 tctatttaat ctgcttttct tgtctaataa atatatatgt aaagtacgct ttttgttgaa
     7981 attttttaaa cctttgttta tttttttttc ttcattccgt aactcttcta ccttctttat
     8041 ttactttcta aaatccaaat acaaaacata aaaataaata aacacagagt aaattcccaa
     8101 attattccat cattaaaaga tacgaggcgc gtgtaagtta caggcaagcg atccgtctaa
     8161 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     8221 c
//
