LOCUS       sequence-217253-        7871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sub2-5 in pRS315.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7871)
  AUTHORS   Kistler AL, Guthrie C
  TITLE     Deletion of MUD2, the yeast homolog of U2AF65, can bypass the 
            requirement for sub2, an essential spliceosomal ATPase.
  JOURNAL   Genes Dev. 2001 Jan 1;15(1):42-9.
  PUBMED    11156604
REFERENCE   2  (bases 1 to 7871)
  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"; journalName: "Genes 
            Dev."; date: "2001-01-1"; volume: "15(1)"; pages: "42-9"
COMMENT     Sequence Label: sub2-5 pRS315
FEATURES             Location/Qualifiers
     source          1..7871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        240..258
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2250..2269)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2251..2269)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2276..2293)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2276..2292)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2285..2307)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2433..2888
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2520..2539)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2730..2751
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3189..3596
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3597..4691
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(5185..5204)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5304..5326
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5364..5382)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5424..5927
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5964..6068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6069..6137,6138..6929)
                     /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(6287..6306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7100..7688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7589..7608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7842..7859
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctcgaa
      241 attaaccctc actaaaggga acaaaagctg gtaccgggcc ccccctcgag gtcgacaaca
      301 cgccgaataa acaaagggtt tcttcaatac ctgatatagt agctagaggg cagaagtttg
      361 aggtgatatc tttctgagtg ttgttatttt tgtaagtcgt aacacggaac tgtttgcctg
      421 ttggaatttt acgttactat tctcatcggt ataactagct tatattcact ttggcaatag
      481 gttaaaaaaa gctcgttcat ttttttaaaa acagcaacga tgtcacacga aggtgaagaa
      541 gatttattgg agtactccga taacgaacaa gaaattcaaa ttgatgcctc taaggccgct
      601 gaagctggag aaactggcgc cgccacttct gcaactgaag gtgataataa taacaacact
      661 gcagctggcg acaagaaagg ttcctatgtt ggtatccatt ccaccggttt caaagatttc
      721 ttgctaaagc cagaactatc aagagccatc attgactgtg gttttgaaca tccttctgag
      781 gtccagcaac ataccattcc tcagtcaatt catggtaccg atgtcttgtg tcaagcaaag
      841 tctggtttag gtaagacagc tgtctttgtc ttatccactc tgcaacaatt ggaccctgtt
      901 ccaggtgaag ttgccgttgt tgtcatttgt aatgctagag aactggccta tcaaattcgt
      961 aacgagtatt tgagattttc caaatatatg ccagacgtta agacagcagt cttttacggt
     1021 ggtactccaa tttctaagga tgctgaactt ttaaagaata aagatactgc tcctcacatt
     1081 gttgttgcaa ctccaggtcg tttaaaagcg ttagtgagag aaaaatacat tgatttgtca
     1141 cacgtcaaga actttgtcat agatgaatgt gataaggttt tagaagaatt agacatgaga
     1201 agagatgtgc aagaaatttt cagagctact ccaagagaca aacaagtcat gatgttttca
     1261 gccacacttt ctcaagaaat tagaccaatt tgtagacgct tcttacagaa tccattggaa
     1321 attttcgtcg atgatgaagc taagctaacc ttgcacgggt tgcaacaata ttatatcaaa
     1381 ttagaagaac gtgaaaagaa ccgtaaattg gctcaattat tagatgattt ggagttcaat
     1441 cgagtcatta ttttcgttaa atctaccaca agagctaatg agttgaccaa actattaaac
     1501 gcctctaact ttccggctat caccgttcat ggtcatatga aacaagaaga acgtattgct
     1561 cgttacaagg ctttcaaaga ttttgaaaaa cgtatttgcg tgtccacaga tgtttttggt
     1621 agaggtatcg atattgaacg tattaattta gccatcaatt acgatttgac taatgaagct
     1681 gaccaatatt tacatcgtgt cggtagagct ggtagatttg gtactaaggg tttggctatt
     1741 tcattcgttt cttcaaagga agatgaggaa gtcttggcta agatccaaga gcgtttcgat
     1801 gtcaaaatcg ctgaattccc agaagaaggc attgatccgt ccacttattt gaataattaa
     1861 taaaggaaaa aagatacgtt tttatataga ttatataaag attttgtatt attcaaacga
     1921 gcaaaacaaa caaaaaaggg gaagtggaaa aaaattagca gttatagcga aagattgagt
     1981 ttttgttgga aaatgaacgt agtgtagtca tttcgtgaac gcacaagaca attctattgg
     2041 gcatcacatg gcaaagcatg ataaggaaat gaagttttgg gaatgattct aaattaacgt
     2101 tttacccgtt ataggaagat atacagtata tgtaagtaaa aagtttttta atacaataga
     2161 tgtgcaaaga aaagtgaaga gatttctgga aaaaatcttg aaattttctt aacggtaaga
     2221 tttttggaat ctagagcggc cccaattcgc cctatagtga gtcgtattac aattcactgg
     2281 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     2341 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     2401 cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc
     2461 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2521 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2581 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2641 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2701 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2761 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2821 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     2881 tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     2941 tcgaccggtc gaggagaact tctagtatat ctacatacct aatattattg ccttattaaa
     3001 aatggaatcc caacaattac atcaaaatcc acattctctt caaaatcaat tgtcctgtac
     3061 ttccttgttc atgtgtgttc aaaaacgtta tatttatagg ataattatac tctatttctc
     3121 aacaagtaat tggttgtttg gccgagcggt ctaaggcgcc tgattcaaga aatatcttga
     3181 ccgcagttaa ctgtgggaat actcaggtat cgtaagatgc aagagttcga atctcttagc
     3241 aaccattatt tttttcctca acataacgag aacacacagg ggcgctatcg cacagaatca
     3301 aattcgatga ctggaaattt tttgttaatt tcagaggtcg cctgacgcat ataccttttt
     3361 caactgaaaa attgggagaa aaaggaaagg tgagagcgcc ggaaccggct tttcatatag
     3421 aatagagaag cgttcatgac taaatgcttg catcacaata cttgaagttg acaatattat
     3481 ttaaggacct attgtttttt ccaataggtg gttagcaatc gtcttacttt ctaacttttc
     3541 ttacctttta catttcagca atatatatat atatatttca aggatatacc attctaatgt
     3601 ctgcccctaa gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa atcacagccg
     3661 aagccattaa ggttcttaaa gctatttctg atgttcgttc caatgtcaag ttcgatttcg
     3721 aaaatcattt aattggtggt gctgctatcg atgctacagg tgttccactt ccagatgagg
     3781 cgctggaagc ctccaagaag gctgatgccg ttttgttagg tgctgtgggt ggtcctaaat
     3841 ggggtaccgg tagtgttaga cctgaacaag gtttactaaa aatccgtaaa gaacttcaat
     3901 tgtacgccaa cttaagacca tgtaactttg catccgactc tcttttagac ttatctccaa
     3961 tcaagccaca atttgctaaa ggtactgact tcgttgttgt cagagaatta gtgggaggta
     4021 tttactttgg taagagaaag gaagacgatg gtgatggtgt cgcttgggat agtgaacaat
     4081 acaccgttcc agaagtgcaa agaatcacaa gaatggccgc tttcatggcc ctacaacatg
     4141 agccaccatt gcctatttgg tccttggata aagctaatgt tttggcctct tcaagattat
     4201 ggagaaaaac tgtggaggaa accatcaaga acgaattccc tacattgaag gttcaacatc
     4261 aattgattga ttctgccgcc atgatcctag ttaagaaccc aacccaccta aatggtatta
     4321 taatcaccag caacatgttt ggtgatatca tctccgatga agcctccgtt atcccaggtt
     4381 ccttgggttt gttgccatct gcgtccttgg cctctttgcc agacaagaac accgcatttg
     4441 gtttgtacga accatgccac ggttctgctc cagatttgcc aaagaataag gtcaacccta
     4501 tcgccactat cttgtctgct gcaatgatgt tgaaattgtc attgaacttg cctgaagaag
     4561 gtaaggccat tgaagatgca gttaaaaagg ttttggatgc aggtatcaga actggtgatt
     4621 taggtggttc caacagtacc accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa
     4681 tccttgctta aaaagattct ctttttttat gatatttgta cataaacttt ataaatgaaa
     4741 ttcataatag aaacgacacg aaattacaaa atggaatatg ttcatagggt agacgaaact
     4801 atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatgta aaggaataca
     4861 ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa aaagaattgc actttaacat
     4921 taatattgac aaggaggagg gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa
     4981 ctatgattcc taatttatat attggaggat tttctctaaa aaaaaaaaaa tacaacaaat
     5041 aaaaaacact caatgacctg accatttgat ggagtttaag tcaatacctt cttgaaccat
     5101 ttcccataat ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg ccttaacgac
     5161 gtagtcgata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     5221 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     5281 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5341 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     5401 gataataatg gtttcttagg acggatcgct tgcctgtaac ttacacgcgc ctcgtatctt
     5461 ttaatgatgg aataatttgg gaatttactc tgtgtttatt tatttttatg ttttgtattt
     5521 ggattttaga aagtaaataa agaaggtaga agagttacgg aatgaagaaa aaaaaataaa
     5581 caaaggttta aaaaatttca acaaaaagcg tactttacat atatatttat tagacaagaa
     5641 aagcagatta aatagatata cattcgatta acgataagta aaatgtaaaa tcacaggatt
     5701 ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc ggcattaata cctgagagca
     5761 ggaagagcaa gataaaaggt agtatttgtt ggcgatcccc ctagagtctt ttacatcttc
     5821 ggaaaacaaa aactattttt tctttaattt ctttttttac tttctatttt taatttatat
     5881 atttatatta aaaaatttaa attataatta tttttatagc acgtgatgaa aaggacccag
     5941 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6001 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6061 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6121 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6181 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6241 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6301 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     6361 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     6421 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     6481 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     6541 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     6601 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     6661 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     6721 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     6781 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6841 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6901 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6961 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7021 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7081 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7141 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7201 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     7261 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7321 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     7381 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7441 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7501 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7561 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7621 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7681 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7741 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     7801 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     7861 aagcggaaga g
//
