LOCUS       sequence-214667-        8375 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  HA-SNU114.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8375)
  AUTHORS   Brenner TJ, Guthrie C
  TITLE     Genetic analysis reveals a role for the C terminus of the 
            Saccharomyces cerevisiae GTPase Snu114 during spliceosome 
            activation.
  JOURNAL   Genetics. 2005 Jul;170(3):1063-80. doi: 10.1534/genetics.105.042044.
            Epub 2005 May 23.
  PUBMED    15911574
REFERENCE   2  (bases 1 to 8375)
  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"; doi: 
            "10.1534/genetics.105.042044"; journalName: "Genetics"; date: 
            "2005-07"; volume: "170"; issue: "3"; pages: "1063-80"
COMMENT     Sequence Label: pRS314 HA-SNU114
FEATURES             Location/Qualifiers
     source          1..8375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     30..47
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    164..185
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(200..205,206..223,224..230)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    238..254
                     /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)."
     primer_bind     243..265
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     262..278
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     262..278
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        299..317
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             670..759
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(4032..4051)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4033..4051)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4058..4075)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4058..4074)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4067..4089)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4215..4670
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4302..4321)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4512..4533
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4773..5447)
                     /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"
     promoter        complement(5448..5728)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5749..5768)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5868..5890
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5928..5946)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5987..6490
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6527..6631
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6632..6700,6701..7492)
                     /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     complement(6850..6869)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7663..8251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8152..8171
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
       61 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
      121 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
      181 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
      241 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctcgaaa
      301 ttaaccctca ctaaagggaa caaaagctgg taccgggccc cccctcgagg tagcgtatac
      361 tctggcattt ttcatttatt tgacttattt ttatttattt tattgtacaa tggcaaacaa
      421 atacacggac tggtatgttc ttcgtcaaat tgatgatctg ttcttttggc ttatccttat
      481 gggatgatta atgattgtta aagcctcatt gcggaaattt taatgccgga aaggcctgaa
      541 aaatttcaaa gtggcagaaa acaaggcttt ggggcattac ttgccaacgg ctgacgattg
      601 ctagctgctt ttacagaagc gatacacaat agctcctaac atagcaaaat ggaattcggc
      661 cgcatctttt acccatacga tgttcctgac tatgcgggct atccctatga cgtcccggac
      721 tatgcaggat cctatccata tgacgttcca gattacgctg ctcagtgcag atctgaaggt
      781 gacgatttat tcgatgagtt tggaaatttg atcggagttg atccttttga ctcggatgaa
      841 gaagaaagtg tgctggatga gcaagaacaa tatcaaacca atacgtttga ggggagcggc
      901 aacaacaatg agattgagag cagacaactt acctcgttag gaagtaaaaa ggaacttgga
      961 atttccttag agcaccctta tggtaaggaa gtggaagtat taatggaaac taagaacaca
     1021 cagtcaccac agactccact ggtagagccg gttacagaac gaacaaagtt gcaagagcat
     1081 acaattttta cacaattaaa aaaaaacatt ccgaagacta gatataatcg agactatatg
     1141 ttatcaatgg ctaacattcc ggagaggata ataaatgttg gtgtcattgg acctcttcac
     1201 tcaggtaaga cctctttgat ggatctccta gtaatcgact cacacaaacg cataccagat
     1261 atgtctaaga atgtagaact tggatggaaa ccgctaagat acctggataa tttgaaacaa
     1321 gagattgacc ggggtctttc cattaaactt aatggttcca ctctactctg caccgattta
     1381 gaatccaagt ctagaatgat aaactttttg gatgcaccag ggcatgttaa ttttatggat
     1441 gaaacggcag tcgcacttgc agcaagtgat ctggttttaa tcgtgataga tgttgtagaa
     1501 ggtgttacat ttgtggtaga gcaattgata aaacaaagta ttaaaaataa tgtagcaatg
     1561 tgttttgtta taaataaact cgacagattg attctagatt taaagttgcc accaatggac
     1621 gcatacttga agttaaatca tataattgca aatatcaatt cctttacgaa gggaaacgtg
     1681 ttttcaccaa tagacaataa cataatattt gcgtctacaa agctgggctt cacattcact
     1741 atcaaggaat ttgtttctta ttattacgcc cactccatac cttcttctaa gatagacgac
     1801 tttaccacaa ggctatgggg tagcgtatat taccataaag gtaatttcag gacaaaacca
     1861 tttgaaaatg tagagaagta cccgactttc gttgaattta tcctcatccc gctttataaa
     1921 atattttcct acgctttatc tatggagaaa gataaattaa agaatttgtt aaggtctaac
     1981 tttagagtta atttaagtca agaggccctc cagtacgatc cgcaaccttt tttaaaacac
     2041 gttctacagt taatatttag gcaacaaacc ggtcttgtag atgcgataac cagatgttat
     2101 caaccatttg aattattcga caataagaca gctcatcttt caattccagg aaaatctaca
     2161 ccagaaggca ctctttgggc tcatgtactg aaaactgtgg attacggtgg agcagagtgg
     2221 tcccttgtac ggatatactc tggacttttg aagagagggg atacagtacg tatacttgat
     2281 acatcacaat ctgaatctcg ccaaaaaaga cagttgcatg atatctcaaa gacggaaacc
     2341 tccaacgaag atgaagatga agatgacgaa acgccttcat gtgaggtcga agaaattggc
     2401 ttattaggtg gcagatatgt ctatccggtt catgaagcac acaaaggaca gatagtatta
     2461 ataaaaggta tttcaagtgc atatattaaa tccgcgacgt tgtactctgt aaagagtaag
     2521 gaggacatga aacagctaaa atttttcaaa cctttagatt atattaccga ggcagttttc
     2581 aaaattgttc ttcaaccttt actgccaagg gaattaccca agttattgga tgccctgaac
     2641 aaaatttcga aatactaccc aggcgttatt attaaagttg aggaatctgg agaacatgta
     2701 atactaggaa atggagagct gtatatggac tgtttgttgt atgatctaag agcgagttat
     2761 gcaaagatag aaattaaaat atctgatcct ttaactgttt tttctgaaag ttgctcaaat
     2821 gaatcgttcg cttcaatacc tgtaagtaat tcgatatctc gccttggtga ggaaaattta
     2881 ccaggtctgt cgattagtgt agccgctgaa ccaatggact ctaaaatgat tcaggatttg
     2941 agcaggaaca cattgggaaa gggtcaaaac tgcttggata ttgacggaat aatggacaac
     3001 ccaaggaaac tatccaaaat attgagaacc gagtatgggt gggattcgtt ggcatcaaga
     3061 aacgtttggt ctttttataa tggcaacgtg ttgattaatg atactttgcc agatgaaatc
     3121 agtcctgaat tattatccaa atataaggaa caaataatac aaggatttta ctgggctgta
     3181 aaagaggggc ctttggcaga agaaccaatt tatggtgtac agtataaatt gttatcgatc
     3241 tcagtacctt ccgacgtaaa cattgacgtt atgaaaagtc aaattattcc gctaatgaaa
     3301 aaagcctgtt acgttggctt actgacagca atcccaattt tattggaacc catctatgaa
     3361 gttgacatca cggtccacgc ccccttgctg ccaatagtag aggaacttat gaagaagaga
     3421 cgtggaagca ggatatacaa aacaataaaa gtggcaggga caccattgtt ggaggttcgt
     3481 ggacaagttc cggttattga atctgcagga ttcgagacag atttgagatt atctacgaat
     3541 ggtcttggta tgtgtcagct gtacttttgg cacaagatat ggaggaaggt tcctggtgat
     3601 gttttggata aagatgcgtt tattccaaaa ttgaaacccg cacctatcaa cagtttaagt
     3661 cgtgatttcg tgatgaaaac aagaaggcgg aagggtattt ctacaggtgg atttatgtca
     3721 aatgatggtc ctacgcttga aaagtatata agcgctgaat tatacgctca attaagagaa
     3781 aatggcttag taccgtgaaa atcttggcgc ataagaatta agcaatatgt ccacaatatt
     3841 tttattcagc agaatttaca tatgacttcg tatttagtga tataactact catcgggact
     3901 gaaaattttt tatttctgcc taaatacaga tgagatgagc tttatagaat tttacgctgt
     3961 taaaaagtta aacggtaaca aagcacagtt agttgaacaa cggtatatag agaatggcga
     4021 gctccaattc gccctatagt gagtcgtatt acaattcact ggccgtcgtt ttacaacgtc
     4081 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     4141 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     4201 tgaatggcga atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4261 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     4321 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4381 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     4441 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4501 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     4561 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4621 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg
     4681 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagatcggca agtgcacaaa
     4741 caatacttaa ataaatacta ctcagtaata acctatttct tagcattttt gacgaaattt
     4801 gctattttgt tagagtcttt tacaccattt gtctccacac ctccgcttac atcaacacca
     4861 ataacgccat ttaatctaag cgcatcacca acattttctg gcgtcagtcc accagctaac
     4921 ataaaatgta agctttcggg gctctcttgc cttccaaccc agtcagaaat cgagttccaa
     4981 tccaaaagtt cacctgtccc acctgcttct gaatcaaaca agggaataaa cgaatgaggt
     5041 ttctgtgaag ctgcactgag tagtatgttg cagtcttttg gaaatacgag tcttttaata
     5101 actggcaaac cgaggaactc ttggtattct tgccacgact catctccatg cagttggacg
     5161 atatcaatgc cgtaatcatt gaccagagcc aaaacatcct ccttaggttg attacgaaac
     5221 acgccaacca agtatttcgg agtgcctgaa ctatttttat atgcttttac aagacttgaa
     5281 attttccttg caataaccgg gtcaattgtt ctctttctat tgggcacaca tataataccc
     5341 agcaagtcag catcggaatc tagagcacat tctgcggcct ctgtgctctg caagccgcaa
     5401 actttcacca atggaccaga actacctgtg aaattaataa cagacatact ccaagctgcc
     5461 tttgtgtgct taatcacgta tactcacgtg ctcaatagtc accaatgccc tccctcttgg
     5521 ccctctcctt ttcttttttc gaccgaatta attcttaatc ggcaaaaaaa gaaaagctcc
     5581 ggatcaagat tgtacgtaag gtgacaagct atttttcaat aaagaatatc ttccactact
     5641 gccatctggc gtcataactg caaagtacac atatattacg atgctgttct attaaatgct
     5701 tcctatatta tatatatagt aatgtcgtga tctatggtgc actctcagta caatctgctc
     5761 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5821 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5881 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     5941 cctattttta taggttaatg tcatgataat aatggtttct tagacggatc gcttgcctgt
     6001 aacttacacg cgcctcgtat cttttaatga tggaataatt tgggaattta ctctgtgttt
     6061 atttattttt atgttttgta tttggatttt agaaagtaaa taaagaaggt agaagagtta
     6121 cggaatgaag aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta
     6181 catatatatt tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa
     6241 gtaaaatgta aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa
     6301 ttcggcatta atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc
     6361 cccctagagt cttttacatc ttcggaaaac aaaaactatt ttttctttaa tttctttttt
     6421 tactttctat ttttaattta tatatttata ttaaaaaatt taaattataa ttatttttat
     6481 agcacgtgat gaaaaggacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6541 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6601 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6661 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6721 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6781 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6841 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6901 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6961 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     7021 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     7081 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     7141 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     7201 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     7261 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     7321 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7381 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7441 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7501 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7561 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7621 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7681 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7741 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7801 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7861 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7921 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7981 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     8041 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     8101 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     8161 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     8221 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     8281 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     8341 ggataaccgt attaccgcct ttgagtgagc tgata
//
