LOCUS       sequence-214709-        6946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prp40 expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6946)
  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 6946)
  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: Prp40-pET21b
FEATURES             Location/Qualifiers
     source          1..6946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     148..167
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        148..166
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    167..191
                     /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)."
     RBS             206..228
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             236..268
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             2033..2050
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2103..2121)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2117..2164
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2201..2656
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2288..2307)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2498..2519
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2682..2786
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2787..2855,2856..3647)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3005..3024)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3818..4406
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4307..4326
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4560..4577
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4592..4731
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4657..4676)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4817..4839
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4833..5024)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5833..6915)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6877..6896
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
       61 gccgttgagc accgccgccg caaggaatgg tgcgcccagc ccgcctaatg agcgggcttt
      121 tttttgggga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
      181 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatggc
      241 tagcatgact ggtggacagc aaatgggtcg ggatcctatg tctatttgga aggaagctaa
      301 agatgcaagt ggaaggatat attattacaa tactttgaca aagaaatcta cgtgggagaa
      361 gcccaaggaa ctaatttctc aggaggagct acttcttcga gaaaatggct ggaaggcggc
      421 caagacggca gacggcaaag tatactatta taatccaaca acaagagaaa ccagctggac
      481 tattccggcc ttcgagaaga aagtagaacc catcgcagaa caaaaacatg atacagtatc
      541 ccatgcacag gttaatggaa atagaatagc ccttacggct ggagaaaaac aagagccggg
      601 acgaactata aacgaggagg aaagccaata tgctaataac tctaaactgc ttaatgtcag
      661 gagaaggact aaagaagaag cagagaagga atttattacc atgctgaagg aaaatcaagt
      721 agactctact tggtcattca gtagaattat ttcagaactg gggaccagag atccaaggta
      781 ttggatggtc gatgatgacc ccttatggaa gaaagaaatg tttgagaaat atctttccaa
      841 tagatcagcc gatcaacttc ttaaggaaca caatgaaaca agcaaattca aagaagcctt
      901 tcagaaaatg ttgcaaaaca attctcatat aaaatattac acccgttggc ctaccgcaaa
      961 gagactaatt gccgacgaac caatatacaa acactccgtg gtcaatgaaa agacaaagag
     1021 acagaccttt caagattata tagataccct catcgacact cagaaagaat caaaaaaaaa
     1081 attgaaaaca caggccctaa aagaactaag agagtattta aacggtatta taacaacatc
     1141 atcctctgaa actttcataa cctggcagca gcttttaaat cactatgttt ttgacaagag
     1201 taagagatat atggcgaacc ggcacttcaa agtcttaacc cacgaagatg ttttaaacga
     1261 gtatctgaaa atagtaaata cgattgaaaa cgatcttcaa aacaaactaa atgagctccg
     1321 actgcgcaat tataccagag accgtattgc tagagataac tttaaaagct tattaagaga
     1381 agtgccaatc aaaatcaaag caaatactag atggtcagat atttatcctc atataaagtc
     1441 tgatccgcgc tttttacata tgcttggaag gaatggctcg tcctgccttg atttattttt
     1501 agattttgtt gatgagcaaa ggatgtacat ctttgcacaa agatcaatag cccaacagac
     1561 gttgatagat caaaattttg aatggaatga taccgatagc gacgagatca ccaagcaaaa
     1621 catagaaaaa gttctggaaa atgaccggaa atttgacaag gtggataaag aagacatcag
     1681 tttgattgtt gatggtttga taaagcaaag aaacgaaaag atacaacaga aactccaaaa
     1741 tgagcgtagg atattggagc aaaagaagca ctatttttgg ttacttttgc aaaggacata
     1801 tacaaaaacc ggtaagccca agcctagtac gtgggattta gcttccaaag agcttggcga
     1861 atctcttgaa tacaaggcac taggcgatga agataacata agaagacaaa ttttcgagga
     1921 ttttaagcct gaaagctctg caccgactgc cgaaagcgct actgcaaact taacgttgac
     1981 cgcgtcaaaa aagaggcatt taactccggc tgtggaattg gactatctcg agcaccacca
     2041 ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     2101 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     2161 tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca
     2221 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     2281 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2341 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2401 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2461 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2521 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     2581 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2641 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2701 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2761 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     2821 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     2881 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     2941 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     3001 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3061 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3121 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3181 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     3241 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     3301 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     3361 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     3421 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     3481 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     3541 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     3601 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     3661 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     3721 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     3781 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     3841 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     3901 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     3961 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     4021 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     4081 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     4141 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     4201 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     4261 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     4321 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     4381 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     4441 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     4501 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     4561 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     4621 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     4681 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     4741 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4801 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4861 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     4921 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     4981 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     5041 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     5101 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     5161 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     5221 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     5281 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     5341 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     5401 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     5461 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     5521 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     5581 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     5641 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     5701 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     5761 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     5821 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5881 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     5941 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     6001 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     6061 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     6121 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     6181 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     6241 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     6301 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     6361 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     6421 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     6481 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     6541 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     6601 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     6661 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     6721 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     6781 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     6841 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     6901 cgttactggt ttcacattca ccaccctgaa ttgactctct tccggg
//
