LOCUS       sequence-217158-        8346 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pRS316-SNU114.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8346)
  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 8346)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pRS316-SNU114
FEATURES             Location/Qualifiers
     source          1..8346
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    63..84
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(99..104,105..122,123..129)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    137..153
                     /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     142..164
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     161..177
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     161..177
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        198..216
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3807..3826)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3808..3826)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3833..3850)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3833..3849)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3842..3864)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3990..4445
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4077..4096)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4287..4308
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4576..5379)
                     /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"
     promoter        complement(5380..5595)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(5619..5638)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5738..5760
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5798..5816)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5857..6360
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6397..6501
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6502..6570,6571..7362)
                     /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(6720..6739)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7533..8121
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8022..8041
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8275..8292
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
       61 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      121 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
      181 gattacgcca agctcgaaat taaccctcac taaagggaac aaaagctggt accgggcccc
      241 cctcgaggta gcgtatactc tggcattttt catttatttg acttattttt atttatttta
      301 ttgtacaatg gcaaacaaat acacggactg gtatgttctt cgtcaaattg atgatctgtt
      361 cttttggctt atccttatgg gatgattaat gattgttaaa gcctcattgc ggaaatttta
      421 atgccggaaa ggcctgaaaa atttcaaagt ggcagaaaac aaggctttgg ggcattactt
      481 gccaacggct gacgattgct agctgctttt acagaagcga tacacaatag ctcctaacat
      541 agcaaaatgg aaggtgacga tttattcgat gagtttggaa atttgatcgg agttgatcct
      601 tttgactcgg atgaagaaga aagtgtgctg gatgagcaag aacaatatca aaccaatacg
      661 tttgagggga gcggcaacaa caatgagatt gagagcagac aacttacctc gttaggaagt
      721 aaaaaggaac ttggaatttc cttagagcac ccttatggta aggaagtgga agtattaatg
      781 gaaactaaga acacacagtc accacagact ccactggtag agccggttac agaacgaaca
      841 aagttgcaag agcatacaat ttttacacaa ttaaaaaaaa acattccgaa gactagatat
      901 aatcgagact atatgttatc aatggctaac attccggaga ggataataaa tgttggtgtc
      961 attggacctc ttcactcagg taagacctct ttgatggatc tcctagtaat cgactcacac
     1021 aaacgcatac cagatatgtc taagaatgta gaacttggat ggaaaccgct aagatacctg
     1081 gataatttga aacaagagat tgaccggggt ctttccatta aacttaatgg ttccactcta
     1141 ctctgcaccg atttagaatc caagtctaga atgataaact ttttggatgc accagggcat
     1201 gttaatttta tggatgaaac ggcagtcgca cttgcagcaa gtgatctggt tttaatcgtg
     1261 atagatgttg tagaaggtgt tacatttgtg gtagagcaat tgataaaaca aagtattaaa
     1321 aataatgtag caatgtgttt tgttataaat aaactcgaca gattgattct agatttaaag
     1381 ttgccaccaa tggacgcata cttgaagtta aatcatataa ttgcaaatat caattccttt
     1441 acgaagggaa acgtgttttc accaatagac aataacataa tatttgcgtc tacaaagctg
     1501 ggcttcacat tcactatcaa ggaatttgtt tcttattatt acgcccactc cataccttct
     1561 tctaagatag acgactttac cacaaggcta tggggtagcg tatattacca taaaggtaat
     1621 ttcaggacaa aaccatttga aaatgtagag aagtacccga ctttcgttga atttatcctc
     1681 atcccgcttt ataaaatatt ttcctacgct ttatctatgg agaaagataa attaaagaat
     1741 ttgttaaggt ctaactttag agttaattta agtcaagagg ccctccagta cgatccgcaa
     1801 ccttttttaa aacacgttct acagttaata tttaggcaac aaaccggtct tgtagatgcg
     1861 ataaccagat gttatcaacc atttgaatta ttcgacaata agacagctca tctttcaatt
     1921 ccaggaaaat ctacaccaga aggcactctt tgggctcatg tactgaaaac tgtggattac
     1981 ggtggagcag agtggtccct tgtacggata tactctggac ttttgaagag aggggataca
     2041 gtacgtatac ttgatacatc acaatctgaa tctcgccaaa aaagacagtt gcatgatatc
     2101 tcaaagacgg aaacctccaa cgaagatgaa gatgaagatg acgaaacgcc ttcatgtgag
     2161 gtcgaagaaa ttggcttatt aggtggcaga tatgtctatc cggttcatga agcacacaaa
     2221 ggacagatag tattaataaa aggtatttca agtgcatata ttaaatccgc gacgttgtac
     2281 tctgtaaaga gtaaggagga catgaaacag ctaaaatttt tcaaaccttt agattatatt
     2341 accgaggcag ttttcaaaat tgttcttcaa cctttactgc caagggaatt acccaagtta
     2401 ttggatgccc tgaacaaaat ttcgaaatac tacccaggcg ttattattaa agttgaggaa
     2461 tctggagaac atgtaatact aggaaatgga gagctgtata tggactgttt gttgtatgat
     2521 ctaagagcga gttatgcaaa gatagaaatt aaaatatctg atcctttaac tgttttttct
     2581 gaaagttgct caaatgaatc gttcgcttca atacctgtaa gtaattcgat atctcgcctt
     2641 ggtgaggaaa atttaccagg tctgtcgatt agtgtagccg ctgaaccaat ggactctaaa
     2701 atgattcagg atttgagcag gaacacattg ggaaagggtc aaaactgctt ggatattgac
     2761 ggaataatgg acaacccaag gaaactatcc aaaatattga gaaccgagta tgggtgggat
     2821 tcgttggcat caagaaacgt ttggtctttt tataatggca acgtgttgat taatgatact
     2881 ttgccagatg aaatcagtcc tgaattatta tccaaatata aggaacaaat aatacaagga
     2941 ttttactggg ctgtaaaaga ggggcctttg gcagaagaac caatttatgg tgtacagtat
     3001 aaattgttat cgatctcagt accttccgac gtaaacattg acgttatgaa aagtcaaatt
     3061 attccgctaa tgaaaaaagc ctgttacgtt ggcttactga cagcaatccc aattttattg
     3121 gaacccatct atgaagttga catcacggtc cacgccccct tgctgccaat agtagaggaa
     3181 cttatgaaga agagacgtgg aagcaggata tacaaaacaa taaaagtggc agggacacca
     3241 ttgttggagg ttcgtggaca agttccggtt attgaatctg caggattcga gacagatttg
     3301 agattatcta cgaatggtct tggtatgtgt cagctgtact tttggcacaa gatatggagg
     3361 aaggttcctg gtgatgtttt ggataaagat gcgtttattc caaaattgaa acccgcacct
     3421 atcaacagtt taagtcgtga tttcgtgatg aaaacaagaa ggcggaaggg tatttctaca
     3481 ggtggattta tgtcaaatga tggtcctacg cttgaaaagt atataagcgc tgaattatac
     3541 gctcaattaa gagaaaatgg cttagtaccg tgaaaatctt ggcgcataag aattaagcaa
     3601 tatgtccaca atatttttat tcagcagaat ttacatatga cttcgtattt agtgatataa
     3661 ctactcatcg ggactgaaaa ttttttattt ctgcctaaat acagatgaga tgagctttat
     3721 agaattttac gctgttaaaa agttaaacgg taacaaagca cagttagttg aacaacggta
     3781 tatagagaat ggcgagctcc aattcgccct atagtgagtc gtattacaat tcactggccg
     3841 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     3901 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     3961 aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc
     4021 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     4081 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     4141 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     4201 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4261 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4321 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4381 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     4441 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataggg
     4501 taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac atgcatttac
     4561 ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt cccagcctgc
     4621 ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata gtcctcttcc
     4681 aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat actgttgacc
     4741 caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc aatcgtaacc
     4801 ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat ctttgtcgct
     4861 cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc ccttgcatga
     4921 caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg ccgcctgctt
     4981 caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt ctgcccattc
     5041 tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa tgtcagcaaa
     5101 ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta gcggcttaac
     5161 tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta aacaaatttt
     5221 gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat ccaatgaagc
     5281 acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa caggactagg
     5341 atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc gtttcctgca
     5401 ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt cttcaacact
     5461 acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt ccttctgttc
     5521 ggagattacc gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag aataaaaaaa
     5581 aaatgatgaa ttgaaaaggt ggtatggtgc actctcagta caatctgctc tgatgccgca
     5641 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     5701 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     5761 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     5821 taggttaatg tcatgataat aatggtttct tagacggatc gcttgcctgt aacttacacg
     5881 cgcctcgtat cttttaatga tggaataatt tgggaattta ctctgtgttt atttattttt
     5941 atgttttgta tttggatttt agaaagtaaa taaagaaggt agaagagtta cggaatgaag
     6001 aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta catatatatt
     6061 tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa gtaaaatgta
     6121 aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa ttcggcatta
     6181 atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc cccctagagt
     6241 cttttacatc ttcggaaaac aaaaactatt ttttctttaa tttctttttt tactttctat
     6301 ttttaattta tatatttata ttaaaaaatt taaattataa ttatttttat agcacgtgat
     6361 gaaaaggacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6421 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6481 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6541 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6601 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6661 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6721 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     6781 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6841 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6901 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6961 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     7021 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     7081 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7141 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7201 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7261 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7321 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7381 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7441 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7501 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7561 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7621 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7681 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7741 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7801 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7861 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7921 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7981 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     8041 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     8101 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     8161 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     8221 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     8281 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     8341 ccgatt
//
