LOCUS       sequence-217743-        7105 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Prp39 expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7105)
  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 7105)
  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: Prp39-pET21b
FEATURES             Location/Qualifiers
     source          1..7105
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     211..230
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        211..229
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    230..254
                     /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             269..291
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             299..331
                     /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             2255..2272
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2325..2343)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2339..2386
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2423..2878
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2510..2529)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2720..2741
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2904..3008
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3009..3077,3078..3869)
                     /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(3227..3246)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4040..4628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4529..4548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4782..4799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4814..4953
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4879..4898)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5039..5061
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5055..5246)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6055..7105,1..32))
                     /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     join(7099..7105,1..13)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tctgcgacat cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc
       61 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
      121 gaggccgttg agcaccgccg ccgcaaggaa tggtgcgccc agcccgccta atgagcgggc
      181 ttttttttgg ggatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
      241 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat
      301 ggctagcatg actggtggac agcaaatggg tcgggatccg aattcgagct ccgtcgacaa
      361 gcttccagat gaaacaaatt ttacaataga agatattgag cctaggcctg atgcccttcg
      421 aggtttggac acccagttcc ttcaagacaa cacagcatta gtacaagcat atcgtggttt
      481 agactggtca gatatttctt cgttgaccca aatggtagat gttatagagc aaacagtggt
      541 taaatatggc aatccaaatg attcaatcaa attggccttg gaaactattt tatggcaaat
      601 attgcgtaag taccctttat tgtttggatt ttggaagagg tttgctacta ttgagtacca
      661 gttatttggt ctcaagaaat cgattgctgt tctggccacc tcggtcaaat ggttcccaac
      721 ttctctggag ttatggtgtg attatttgaa tgtgttgtgt gttaataacc cgaacgaaac
      781 ggattttatt aggaataatt ttgaaattgc aaaggatctt attggcaagc agtttctatc
      841 acaccctttt tgggacaagt ttatagaatt tgaagttggg cagaagaatt ggcataacgt
      901 tcaaagaata tacgagtaca ttattgaagt tcctctacat cagtatgcgc ggtttttcac
      961 atcttacaag aagtttttaa atgaaaaaaa tttgaagaca acaaggaata ttgatatagt
     1021 tttacgaaaa acacaaacaa cagtaaatga aatttggcag ttcgaatcga aaatcaaaca
     1081 gcccttcttc aacctgggcc aagtgttaaa tgatgatttg gaaaattggt cgcgatattt
     1141 aaaatttgtg acggatcctt caaaatcctt ggataaagaa ttcgtaatgt ccgtttttga
     1201 cagatgtttg attccttgtc tttaccatga aaatacgtgg atgatgtata ttaagtggct
     1261 aacaaagaaa aatatatccg atgaagtggt agtggatata taccaaaaag ccaatacgtt
     1321 cttaccactg gatttcaaaa ctttgaggta tgattttctt aggtttttga aaagaaaata
     1381 tcggtcaaat aataccttat tcaataacat attcaatgag acagtatcgc gttatttaaa
     1441 gatttggccc aacgacatat tacttatgac ggagtacctt tgtatgttga aaagacattc
     1501 ttttaagaat tccttagatc aatcgccaaa ggaaattcta gagaaacaaa catcgtttac
     1561 taaaatccta gagacatcaa tcaccaacta catcaataat caaattgata ccaaggtgca
     1621 tttacagacc ttaataaatg ataaaaactt aagcatagtc gtggtagaat tgataaaaac
     1681 tacatggctt gtacttaaaa ataatatgca aacaaggaaa tatttcaacc tttatcaaaa
     1741 aaacatcctc attaaaaatt ctgtaccgtt ctggttaact tactacaaat ttgaaaagag
     1801 taatgtaaat tttaccaagc tgaataaatt cattagggag cttggggtag aaatatactt
     1861 gccgactaca gtaatgaatg atattttgac ggattataag actttctatt taactcattc
     1921 aaatatcgtc acgtatgaat cctccattat agattcaaat acatttgacc ctatcttgta
     1981 ccctgagctg aaaatgtcga atccaaaata cgatccagta ttaaatacta cagcaaatgt
     2041 agattggcat aaaaagacgg aatggaagga ggctggtcac atcggcgtaa cgacagaaag
     2101 accgcagata tccaacagta taattgagtg caattcagga actttgattc aaaagcccat
     2161 cagtttgcct aattttagaa acttggagaa aattaatcag gtcaaaatta atgatctata
     2221 cactgaagaa tttcttaagg aaggtaaact cgagcaccac caccaccacc actgagatcc
     2281 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     2341 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     2401 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2461 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2521 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2581 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2641 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2701 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2761 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2821 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     2881 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2941 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3001 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3061 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3121 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3181 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3241 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3301 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3361 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3421 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3481 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3541 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3601 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3661 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3721 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3781 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3841 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3901 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3961 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4021 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4081 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4141 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4201 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4261 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4321 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4381 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4441 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4501 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4561 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4621 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4681 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4741 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4801 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4861 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     4921 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     4981 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     5041 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     5101 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     5161 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     5221 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     5281 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     5341 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     5401 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     5461 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     5521 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     5581 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     5641 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     5701 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     5761 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     5821 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     5881 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     5941 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     6001 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     6061 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6121 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     6181 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     6241 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     6301 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     6361 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     6421 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     6481 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     6541 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     6601 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     6661 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     6721 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     6781 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     6841 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     6901 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     6961 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     7021 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     7081 ggtctgataa gagacaccgg catac
//
