LOCUS       sequence-177966-        8689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  prp16-302 allele in TRP plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8689)
  AUTHORS   Mayerle M, Guthrie C
  TITLE     Prp8 retinitis pigmentosa mutants cause defects in the transition 
            between the catalytic steps of splicing.
  JOURNAL   RNA. 2016 May;22(5):793-809. doi: 10.1261/rna.055459.115. Epub 2016 
            Mar 11.
  PUBMED    26968627
REFERENCE   2  (bases 1 to 8689)
  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: 
            "10.1261/rna.055459.115"; journalName: "RNA"; date: "2016-05"; 
            volume: "22"; issue: "5"; pages: "793-809"
COMMENT     Sequence Label: pRS314-prp16-302
FEATURES             Location/Qualifiers
     source          1..8689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..22
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(37..42,43..60,61..67)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    75..91
                     /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     80..102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     99..115
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     99..115
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4683..4700)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4683..4699)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4692..4714)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     misc_feature    5214..5506
                     /label=CEN4
                     /note="S. cerevisiae CEN4 centromere"
     rep_origin      5507..5960
                     /label=ARS1
                     /note="S. cerevisiae autonomously replicating sequence 
                     ARS1"
     CDS             complement(5799..6473)
                     /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(6474..6575)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(6592..6610)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6678..6782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6783..6851,6852..7643)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7001..7020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7814..8402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8303..8322
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8556..8573
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
       61 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgacatgat
      121 tacgaattcg agctcggtac ccgggctccc attagcagca gcaattggct tctttggtgt
      181 cttcttcttc ttcaaagttt tcttcaattc cgccagcgtg ctttccaaat cgaaattgtt
      241 gtcaaacaaa gctagcttaa gtgaaagatt atcccagcct tggtaatcct gtaactgcgc
      301 ctttagagta ggaaatacct cgttcattag atcatattca tcgtcattca aataatcatc
      361 aaattcacct tcgtcgtgaa agtccggcat gtcgtctgct ccatcgctgt agtcactgta
      421 agccatctcg atagtcaata gttgtcgaag cagaagcctt tgggtgtctc caagctgtac
      481 ttaactcttt ttaaattgct tattggttag ctttcatgtg gtatattcaa gtcacattaa
      541 tgggcgtctc cgtgcattgt gatcaatata tgaatacgaa gtacttatag agtagtattt
      601 acacgattgt tattatattt atatagcatt cacgcctggt accacaggag ttttctacgc
      661 ttacggtcct ctctggctat acgcctcttc tccttccagc gtgattggat ttcttgcaat
      721 ctactttcca taatttgtac atgttgttca ctaggtttcg agaccatgga tacaaataat
      781 ggcgaaatgc catattttgc agccaaaaat ttgcgtgtat acttttgagg attgtcaaga
      841 cgtaacttcc ggatctcatc aatttcttga ggtcccagat gataggtttt gccgttcact
      901 gtgctcttac ctacgagaac atttggacac tgcttcgctt tagtgctttg aacaggcatg
      961 tcaagccctc tacgagggtc actggctgcc ataaagctac gtggaatggt ctcactgtta
     1021 acagatccgg ttgcgatgga ttctgatggt tgatagtaca ttcctggagc atctctgtgc
     1081 tttaaatacc ctttatagtt agatgcgctc ttagtcttgt tgtaaatcgt tgtcacttgt
     1141 gttttgggaa atccaaattg cttccaggca gatccagcag tgtggaacga tctgcagcac
     1201 acacctctgc caatcattct tgtgctattt tactttgtct ttaattttcc attttaactt
     1261 gttgaaagta tctctttttg ccctcttttc tattttctca aataataaaa gcggtgaaag
     1321 ctgaaaaatt catgagatga ctgcagatga taaaacacaa gaaggcccaa agacaataca
     1381 aatacctctg gtatgggtca ttcggggcgt gaggaaagga taaaagacat atttaaggaa
     1441 cttacgtcaa aagaactaac gcctgggttg cttttgacac tgcagaaact agcgcaaaag
     1501 ccgaatacta atctagaaca gtttattgcc agctgtaaag ctttgacgaa actcagcagt
     1561 aacaatccaa tcatttttaa tgaacttctg gagctcctca agaataaatc tgaagaggac
     1621 tcgacaggcc caaagaaaat cgccccttct ataaataaga gaaagaaatt taagatccag
     1681 ttagaccttg acgataatga agatgagcta gattctccag ttcaaaaaaa accagcacct
     1741 acacggacgt tgttcaagag aatagataaa ctaaaggcca aacagttgag gcagtattcg
     1801 cccaccgtga aggatccttc tccaaatagt gaacagcaaa cacagaatgg tcatgctgaa
     1861 acgaaggact atgaaccgac tcgaagcgaa gtagttgagg aggacagaga atggtatgat
     1921 aatgatgatg attatggaaa tttggtgcct gaacctctgt ctgaattgcc tgaggaagcg
     1981 aaactactac ctgtcattag gaatatcgac aacgatgatg ccttacgaaa taccgttcaa
     2041 ttatacccca taccactcaa gcaaagaatg gaatggatcc ctcctttttt atcaaagttt
     2101 gctcttgaaa acaaagtgcc cacttccata attatcgggt ccatctctga gacgtccagc
     2161 caagtatccg ctttatcaat ggtgaatcca tttaggaatc ctgacagtga attttctgcg
     2221 aacgccaaac gaggtagtaa gctggttgcc ttgagaagga taaacatgga acatattcaa
     2281 caatctcgag acaacaccac tgttttgaac actgccatgg gggaagtact aggcttggag
     2341 aataacaata aagcaaaaga taagagcaac caaaagattt gcgacgatac agctcttttc
     2401 acgccatcaa aagatgacat taaacatact aaagagcaac tgcctgtttt ccgctgcaga
     2461 tctcaattgt tatcattgat aagagaaaat caagtagtgg tgataattgg tgaaacgggc
     2521 tcaggtaaaa ccacgcaact tgcacagtat ttatatgaag aaggatatgc caacgatagg
     2581 gggaaatcta ttgttgtcac acagccgaga agagtagcag ccatatccgt tgcaaaaagg
     2641 gttgcaatgg aaatgcaagt tccattgggt aaggaagtag gctattctat aaggtttgaa
     2701 gatgtgacgg attctgagtg tacaaaactt aagtttgtaa cggatggtat tctactcaaa
     2761 gagaccttat tggatgatac actcgataaa tattcgtgtg ttattattga tgaagctcat
     2821 gaaaggtcat taaatacaga cattttacta ggttttttca agattttatt agcacgtaga
     2881 agagatctaa aactcataat cacttcagca acaatgaatg cgaagaagtt ctccgcattt
     2941 tttggtaatg ctcctcagtt cacaattcct ggaagaactt tccccgtaca gaccatatat
     3001 acttccaacc ctgttcaaga ctacgtagaa gctgcagttt ctcaggctgt gaaaattcat
     3061 ttagcaaacg attgttcaag cggtgatatt ctcattttca tgacaggtca agaagatatt
     3121 gaaacaacgt tcgacactct gcaagaaaag tttttgcaag tatattctaa aaagtttggt
     3181 accgcaaact ttgaagagat caatgatatt gaaattttac caatctactc ggctttacct
     3241 gcagatttgc aattcaaaat ttttcaggat ctacatggca ctaaaaggaa aattataatt
     3301 gcgacaaata ttgcagagac ttcattgacc attaaaggta ttagatatgt cattgattgt
     3361 ggttactcta aactgaaagt ttacaatcca aagattgggc ttgatagttt agtaataacg
     3421 cccatatcaa aagctaacgc cgaccaaaga tccggaagag cgagaagaac tgcgccaggc
     3481 actgcatatc gattatacac ggaagataca ttcaaagaag atatgtactt acagacaata
     3541 cctgagatcc aaagaacgaa tttgtcaaac acccttttac tgctaaaatc tctggacgtt
     3601 actgacgaat tgagtaagtt cccgttcatt gataaaccac ccttacaaac atttttatcc
     3661 tcattgtatg agttatggtt cataggggct atagacacta gtggacaatt gactccattg
     3721 ggattacaga tggcaaaatt tcccctgcaa ccttcccttt ccaaaatttt attaattgca
     3781 gttagaaacg gctgcagtga tgaaatgtta acaatagttt ccatgctttc tgttccacaa
     3841 gtcttttata gacccaagga aagacagaaa gaagcagata ttgctagaaa taaatttttt
     3901 atcgcgaaat cagatcactt aacgttactt aatgtttttg aacaatggag ggcaaataat
     3961 tttagttcgc attggtgtaa caaacatttt gttcaataca aatcacttgt cagggcaaga
     4021 gacattcgag accagttgct aacaatacta aaatctcaaa aaattccggt catttcctcc
     4081 ggtaaggact gggatattat taaaaaatgt atttgttcag gatttgctca tcaagcagcc
     4141 aaaataacag gactacgaaa ttacgttcat ttaaaaactg gcgtaagtgt tcaactacac
     4201 cctacaagtg cattacacgg attgggggat ttaccccctt acgtggtata tcatgaatta
     4261 ttaatgacta gcaaagaata catttgttgt gttacttctg tagatccatt ctggttaatg
     4321 gaatatggtg ggttactcta cgacatcaag cgtattaaaa atgatcagga ggccacaact
     4381 acaggcctct ttggtgaaca ctatgaacat actctggaca aggttgaaga tgatattgat
     4441 atcaatatta ggagatgtaa ggatatgaga gacagtgtga tacaggagct caaaatgacc
     4501 gacaattcaa acaaagaaga caaaaagcaa aaaacgaaaa agcaaaatat actgaacggc
     4561 aaagaaaatt caatgaaacc tttcaaaaga aggaagcctt ttttttagtt tatgggaaga
     4621 gcgtgctaat aggcaaaata ttcatatatg ttattatata gttatatgca tgcaagcttg
     4681 gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     4741 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     4801 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     4861 cttacgcatc tgtgcggtat ttcacaccgc ataactagct tttaacttgt atcctaggtt
     4921 atctatgctg tctcaccata gagaatatta cctatttcag aatgtatgtc catgattcgc
     4981 cgggtaaata catataatac acaaatctgg cttaataaag tctataatat atctcataaa
     5041 gaagtgctaa attggctagt gctatatatt tttaagaaaa tttcttttga ctaagtccat
     5101 atcgactttg taaaagttca cattagcata catatattac acgagccaga aatagtaact
     5161 tttgcctaaa tcacaaattg caaaatttaa ttgcttgcaa aaggtcacat gcttataatc
     5221 aactttttta aaaatttaaa atactttttt attttttatt tttaaacata aatgaaataa
     5281 tttatttatt gtttatgatt accgaaacat aaaacctgct caagaaaaag aaactgtttt
     5341 gtccttggaa aaaaagcact acctaggagc ggccaaaatg ccgaggcttt catagcttaa
     5401 actctttaca gaaaataggc attatagatc agttcgagtt ttcttattct tccttccggt
     5461 tttatcgtca cagttttaca gtaaataagt atcacctctt agagttggcg gttgtttgca
     5521 agaccgagaa aaggctagca agaatcgggt cattgtagcg tatgcgcctg tgaacattct
     5581 cttcaacaag tttgattcca ttgcggtgaa atggtaaaag tcaaccccct gcgatgtata
     5641 ttttcctgta caatcaatca aaaagccaaa tgatttagca ttatctttac atcttgttat
     5701 tttacagatt ttatgtttag atcttttatg cttgcttttc aaaaggcctg caggcaagtg
     5761 cacaaacaat acttaaataa atactactca gtaataacct atttcttagc atttttgacg
     5821 aaatttgcta ttttgttaga gtcttttaca ccatttgtct ccacacctcc gcttacatca
     5881 acaccaataa cgccatttaa tctaagcgca tcaccaacat tttctggcgt cagtccacca
     5941 gctaacataa aatgtaagct ttcggggctc tcttgccttc caacccagtc agaaatcgag
     6001 ttccaatcca aaagttcacc tgtcccacct gcttctgaat caaacaaggg aataaacgaa
     6061 tgaggtttct gtgaagctgc actgagtagt atgttgcagt cttttggaaa tacgagtctt
     6121 ttaataactg gcaaaccgag gaactcttgg tattcttgcc acgactcatc tccatgcagt
     6181 tggacgatat caatgccgta atcattgacc agagccaaaa catcctcctt aggttgatta
     6241 cgaaacacgc caaccaagta tttcggagtg cctgaactat ttttatatgc ttttacaaga
     6301 cttgaaattt tccttgcaat aaccgggtca attgttctct ttctattggg cacacatata
     6361 atacccagca agtcagcatc ggaatctaga gcacattctg cggcctctgt gctctgcaag
     6421 ccgcaaactt tcaccaatgg accagaacta cctgtgaaat taataacaga catactccaa
     6481 gctgcctttg tgtgcttaat cacgtatact cacgtgctca atagtcacca atgccctccc
     6541 tcttggccct ctccttttct tttttcgacc gaatttcttg aagacgaaag ggcctcgtga
     6601 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     6661 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     6721 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     6781 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     6841 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     6901 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     6961 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     7021 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     7081 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     7141 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     7201 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     7261 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     7321 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     7381 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     7441 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     7501 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     7561 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     7621 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     7681 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     7741 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     7801 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     7861 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     7921 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     7981 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     8041 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     8101 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     8161 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     8221 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     8281 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     8341 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     8401 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     8461 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     8521 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     8581 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     8641 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaat
//
