LOCUS       sequence-217737-        4573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prp28 expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4573)
  AUTHORS   Price AM, Gornemann J, Guthrie C, Brow DA
  TITLE     An unanticipated early function of DEAD-box ATPase Prp28 during 
            commitment to splicing is modulated by U5 snRNP protein Prp8.
  JOURNAL   RNA. 2014 Jan;20(1):46-60. doi: 10.1261/rna.041970.113. Epub 2013 
            Nov 14.
  PUBMED    24231520
REFERENCE   2  (bases 1 to 4573)
  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.041970.113"; journalName: "RNA"; date: "2014-01"; 
            volume: "20"; issue: "1"; pages: "46-60"
COMMENT     Sequence Label: pRSETA-Prp28
FEATURES             Location/Qualifiers
     source          1..4573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..145
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     230..249
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        230..248
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             280..302
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2080..2100
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2101..2118
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2184..2202)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2198..2245
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2342..2797
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2429..2448)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2639..2660
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2823..2927
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2928..2996,2997..3788)
                     /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(3146..3165)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3959..4547
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4448..4467
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
       61 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
      121 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
      181 ccccgcgcgt tggccgattc attaatgcag gatctcgatc ccgcgaaatt aatacgactc
      241 actataggga gaccacaacg gtttccctct agaaataatt ttgtttaact ttaagaagga
      301 gatatacata tggcaagacc aatcgatgtt agtcagctga ttgctggtat aaacaagaaa
      361 aaaggactag atgaaaatac atcaggtaaa atcagcaagc ctaggttttt gaataaacaa
      421 gaaagatcta agcaagaacg tttgaaggag aacgaagaga gtttgacgcc tacgcaaagt
      481 gatagcgcta aggtggagat taaaaaggta aactctagag atgacagttt tttcaatgaa
      541 acaaatgata agaagagaaa tccatcaaag caaaatggct ctaaatttca cttttcttgg
      601 aatgaaagcg aagatactct gagcggatat gatcctatcg tttcaactag agctatagat
      661 ttgctatgga agggaaagac tcctaaaaat gctgcagaat cttcatatat ggggaagcat
      721 tggactgaga agtccttgca cgaaatgaat gagagagatt ggaggatact aaaagaagac
      781 tacgctattg ttacaaaagg tgggacagta gaaaatccat tgagaaattg ggaagagctc
      841 aatataatac caagggactt gttacgtgtt attatacagg agctgcgatt tccttcacca
      901 actcctattc aaaggattac tattccaaac gtttgcaata tgaaacaata tagagatttt
      961 cttggtgttg cctctacagg ttcgggtaag actttggcct ttgtaatacc gatactaatc
     1021 aagatgagta ggtcacctcc aaggccacca tctttgaaga ttattgatgg ccccaaagcg
     1081 ttaattttag ctccaacaag ggaattagtg cagcaaattc aaaaagaaac acaaaaagtt
     1141 acaaaaatat ggtccaaaga gagcaactat gactgcaaag taatttctat tgttggtgga
     1201 cactccttgg aggaaatttc attttctcta tcggagggtt gtgacatcct ggtggccact
     1261 ccaggacgtt taattgactc tttagaaaat cacttacttg tgatgaaaca ggtggagaca
     1321 ttagtactag atgaagctga caaaatgatt gatttaggtt ttgaggacca agtaacaaac
     1381 attttaacta aagtcgatat aaatgctgac tctgctgtga atagacaaac cctgatgttt
     1441 accgctacaa tgacacccgt tatagaaaaa attgcagcag gatacatgca aaagcctgtt
     1501 tatgcaacca ttggggttga gacgggttct gaacctttga ttcaacaggt tgtggaatat
     1561 gcagataatg acgaagacaa attcaaaaag ttgaagccaa ttgtcgctaa atatgatcca
     1621 ccaataataa tttttatcaa ttacaaacaa actgctgatt ggttagcaga aaagtttcaa
     1681 aaggaaacca acatgaaggt gacgatttta catggttcta aatcgcaaga gcaaagggaa
     1741 cattctttgc aattatttcg caccaacaaa gtccaaatca tgatcgcaac aaacgttgcc
     1801 gcaagaggat tggatatccc taatgtttcc ttggtagtca atttccagat ttccaaaaaa
     1861 atggatgact acatccatag aattggtaga acaggtcgtg cagcaaatga gggcactgct
     1921 gtctcctttg ttagtgccgc tgaagatgaa tcattgatac gtgaacttta caaatacgtc
     1981 aggaaacatg atcccttgaa tagcaatatc ttttccgaag ccgtaaagaa caaatataac
     2041 gtcggcaaac agctcagtaa tgagattata tatggcgccg aaaatcttta ttttcaaggt
     2101 catcatcatc atcatcatta ggaattcgaa gcttgatccg gctgctaaca aagcccgaaa
     2161 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     2221 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat ctggcgtaat
     2281 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     2341 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2401 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2461 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2521 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2581 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2641 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2701 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2761 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     2821 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2881 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2941 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3001 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3061 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3121 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3181 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3241 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3301 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3361 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3421 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3481 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3541 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3601 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3661 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3721 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3781 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3841 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3901 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3961 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     4021 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     4081 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     4141 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4201 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4261 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4321 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4381 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4441 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4501 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4561 cggccttttt acg
//
