LOCUS       sequence-214668-        8252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pRS314 snu114-60.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8252)
  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 8252)
  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 snu114-60
FEATURES             Location/Qualifiers
     source          1..8252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..56
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     290..307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    424..445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(460..465,466..483,484..490)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    498..514
                     /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     503..525
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     522..538
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     522..538
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        559..577
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4169..4188)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4170..4188)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4195..4212)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4195..4211)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4204..4226)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4352..4807
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4439..4458)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4649..4670
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4910..5584)
                     /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(5585..5865)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(5886..5905)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6005..6027
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6065..6083)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    6124..6627
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6664..6768
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6769..6837,6838..7629)
                     /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(6987..7006)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7800..8252,1..136)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
       61 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
      361 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
      421 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
      481 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
      541 tgattacgcc aagctcgaaa ttaaccctca ctaaagggaa caaaagctgg taccgggccc
      601 cccctcgagg tagcgtatac tctggcattt ttcatttatt tgacttattt ttatttattt
      661 tattgtacaa tggcaaacaa atacacggac tggtatgttc ttcgtcaaat tgatgatctg
      721 ttcttttggc ttatccttat gggatgatta atgattgtta aagcctcatt gcggaaattt
      781 taatgccgga aaggcctgaa aaatttcaaa gtggcagaaa acaaggcttt ggggcattac
      841 ttgccaacgg ctgacgattg ctagctgctt ttacagaagc gatacacaat agctcctaac
      901 atagcaaaat ggaaggtgac gatttattcg atgagtttgg aaatttgatc ggagttgatc
      961 cttttgactc ggatgaagaa gaaagtgtgc tggatgagca agaacaatat caaaccaata
     1021 cgtttgaggg gagcggcaac aacaatgaga ttgagagcag acaacttacc tcgttaggaa
     1081 gtaaaaagga acttggaatt tccttagagc acccttatgg taaggaagtg gaagtattaa
     1141 tggaaactaa gaacacacag tcaccacaga ctccactggt agagccggtt acagaacgaa
     1201 caaagttgca agagcataca atttttacac aattaaaaaa aaacattccg aagactagat
     1261 ataatcgaga ctatatgtta tcaatggcta acattccgga gaggataata aatgttggtg
     1321 tcattggacc tcttcactca ggtaagacct ctttgatgga tctcctagta atcgactcac
     1381 acaaacgcat accagatatg tctaagaatg tagaacttgg atggaaaccg ctaagatacc
     1441 tggataattt gaaacaagag attgaccggg gtctttccat taaacttaat ggttccactc
     1501 tactctgcac cgatttagaa tccaagtcta gaatgataaa ctttttggat gcaccagggc
     1561 atgttaattt tatggatgaa acggcagtcg cacttgcagc aagtgatctg gttttaatcg
     1621 tgatagatgt tgtagaaggt gttacatttg tggtagagca attgataaaa caaagtatta
     1681 aaaataatgt agcaatgtgt tttgttataa ataaactcga cagattgatt ctagatttaa
     1741 agttgccacc aatggacgca tacttgaagt taaatcatat aattgcaaat atcaattcct
     1801 ttacgaaggg aaacgtgttt tcaccaatag acaataacat aatatttgcg tctacaaagc
     1861 tgggcttcac attcactatc aaggaatttg tttcttatta ttacgcccac tccatacctt
     1921 cttctaagat agacgacttt accacaaggc tatggggtag cgtatattac cataaaggta
     1981 atttcaggac aaaaccattt gaaaatgtag agaagtaccc gactttcgtt gaatttatcc
     2041 tcatcccgct ttataaaata ttttcctacg ctttatctat ggagaaagat aaattaaaga
     2101 atttgttaag gtctaacttt agagttaatt taagtcaaga ggccctccag tacgatccgc
     2161 aacctttttt aaaacacgtt ctacagttaa tatttaggca acaaaccggt cttgtagatg
     2221 cgataaccag atgttatcaa ccatttgaat tattcgacaa taagacagct catctttcaa
     2281 ttccaggaaa atctacacca gaaggcactc tttgggctca tgtactgaaa actgtggatt
     2341 acggtggagc agagtggtcc cttgtacgga tatactctgg acttttgaag agaggggata
     2401 cagtacgtat acttgataca tcacaatctg aatctcgcca aaaaagacag ttgcatgata
     2461 tctcaaagac ggaaacctcc aacgaagatg aagatgaaga tgacgaaacg ccttcatgtg
     2521 aggtcgaaga aattggctta ttaggtggca gatatgtcta tccggttcat gaagcacaca
     2581 aaggacagat agtattaata aaaggtattt caagtgcata tattaaatcc gcgacgttgt
     2641 actctgtaaa gagtaaggag gacatgaaac agctaaaatt tttcaaacct ttagattata
     2701 ttaccgaggc agttttcaaa attgttcttc aacctttact gccaagggaa ttacccaagt
     2761 tattggatgc cctgaacaaa atttcgaaat actacccagg cgttattatt aaagttgagg
     2821 aatctggaga acatgtaata ctaggaaatg gagagctgta tatggactgt ttgttgtatg
     2881 atctaagagc gagttatgca aagatagaaa ttaaaatatc tgatccttta actgtttttt
     2941 ctgaaagttg ctcaaatgaa tcgttcgctt caatacctgt aagtaattcg atatctcgcc
     3001 ttggtgagga aaatttacca ggtctgtcga ttagtgtagc cgctgaacca atggactcta
     3061 aaatgattca ggatttgagc aggaacacat tgggaaaggg tcaaaactgc ttggatattg
     3121 acggaataat ggacaaccca aggaaactat ccaaaatatt gagaaccgag tatgggtggg
     3181 attcgttggc atcaagaaac gtttggtctt tttataatgg caacgtgttg attaatgaca
     3241 ctttgccaga tgaaatcagt cctgaattat tatccaaata taaggaacaa ataatacaag
     3301 gattttactg ggctgtaaaa gaggggcctt tggcagaaga accaatttat ggtgtacagt
     3361 ataaattgtt atcgatctca gtaccttccg acgtaaacat tgacgttatg aaaagtcaaa
     3421 ttattccgct aatgaaaaaa gcctgttacg ttggcttact gacagcaatc ccaattttat
     3481 tggaacccat ctatgaagtt gacatcacgg tccacgcccc cttgctgcca atagtagagg
     3541 aacttatgaa gaagagacgt ggaagcagga tatacaaaac aataaaagtg gcagggacac
     3601 cattgttgga ggttcgtgga caagttccgg ttattgaatc tgcaggattc gagacagatt
     3661 tgagattatc tacgaatggt cttggtatgt gtcagctgta cttttggcac aagatatgga
     3721 ggtaggttcc tggtgatgtt ttggataaag atgcgtttat tccaaaattg aaacccgcac
     3781 ctatcaacag tttaagtcgt gatttcgtga tgaaaacaag aaggcggaag ggtatttcta
     3841 caggtggatt tatgtcaaat gatggtccta cgcttgaaaa gtatataagc gctgaattat
     3901 acgctcaatt aagagaaaac ggcttagtac cgtgaaaatc ttggcgcata agaattaagc
     3961 aatatgtcca caatattttt attcagcaga atttacatat gacttcgtat ttagtgatat
     4021 aactactcat cgggactgaa aattttttat ttctgcctaa atacagatga gatgagcttt
     4081 atagaatttt acgctgttaa aaagttaaac ggtaacaaag cacagttagt tgaacaacgg
     4141 tatatagaga atggcgagct ccaattcgcc ctatagtgag tcgtattaca attcactggc
     4201 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     4261 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     4321 ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg
     4381 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4441 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4501 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4561 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4621 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4681 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4741 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     4801 acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag
     4861 atcggcaagt gcacaaacaa tacttaaata aatactactc agtaataacc tatttcttag
     4921 catttttgac gaaatttgct attttgttag agtcttttac accatttgtc tccacacctc
     4981 cgcttacatc aacaccaata acgccattta atctaagcgc atcaccaaca ttttctggcg
     5041 tcagtccacc agctaacata aaatgtaagc tttcggggct ctcttgcctt ccaacccagt
     5101 cagaaatcga gttccaatcc aaaagttcac ctgtcccacc tgcttctgaa tcaaacaagg
     5161 gaataaacga atgaggtttc tgtgaagctg cactgagtag tatgttgcag tcttttggaa
     5221 atacgagtct tttaataact ggcaaaccga ggaactcttg gtattcttgc cacgactcat
     5281 ctccatgcag ttggacgata tcaatgccgt aatcattgac cagagccaaa acatcctcct
     5341 taggttgatt acgaaacacg ccaaccaagt atttcggagt gcctgaacta tttttatatg
     5401 cttttacaag acttgaaatt ttccttgcaa taaccgggtc aattgttctc tttctattgg
     5461 gcacacatat aatacccagc aagtcagcat cggaatctag agcacattct gcggcctctg
     5521 tgctctgcaa gccgcaaact ttcaccaatg gaccagaact acctgtgaaa ttaataacag
     5581 acatactcca agctgccttt gtgtgcttaa tcacgtatac tcacgtgctc aatagtcacc
     5641 aatgccctcc ctcttggccc tctccttttc ttttttcgac cgaattaatt cttaatcggc
     5701 aaaaaaagaa aagctccgga tcaagattgt acgtaaggtg acaagctatt tttcaataaa
     5761 gaatatcttc cactactgcc atctggcgtc ataactgcaa agtacacata tattacgatg
     5821 ctgttctatt aaatgcttcc tatattatat atatagtaat gtcgtgatct atggtgcact
     5881 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5941 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     6001 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     6061 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     6121 acggatcgct tgcctgtaac ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg
     6181 gaatttactc tgtgtttatt tatttttatg ttttgtattt ggattttaga aagtaaataa
     6241 agaaggtaga agagttacgg aatgaagaaa aaaaaataaa caaaggttta aaaaatttca
     6301 acaaaaagcg tactttacat atatatttat tagacaagaa aagcagatta aatagatata
     6361 cattcgatta acgataagta aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca
     6421 cagacaagat gaaacaattc ggcattaata cctgagagca ggaagagcaa gataaaaggt
     6481 agtatttgtt ggcgatcccc ctagagtctt ttacatcttc ggaaaacaaa aactattttt
     6541 tctttaattt ctttttttac tttctatttt taatttatat atttatatta aaaaatttaa
     6601 attataatta tttttatagc acgtgatgaa aaggacccag gtggcacttt tcggggaaat
     6661 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     6721 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     6781 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     6841 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6901 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6961 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     7021 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     7081 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     7141 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     7201 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     7261 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     7321 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     7381 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     7441 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     7501 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     7561 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     7621 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     7681 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     7741 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     7801 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     7861 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     7921 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     7981 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     8041 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     8101 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     8161 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     8221 agaaaggcgg acaggtatcc ggtaagcggc ag
//
