LOCUS       sequence-217250-        7871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sub2-1 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: sub2-1 pRS315
FEATURES             Location/Qualifiers
     source          1..7871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     19..36
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    153..174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(189..194,195..212,213..219)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    227..243
                     /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     232..254
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     251..267
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     251..267
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        288..306
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2298..2317)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2299..2317)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2324..2341)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2324..2340)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2333..2355)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2481..2936
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2568..2587)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2778..2799
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3237..3644
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3645..4739
                     /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(5233..5252)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5352..5374
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5412..5430)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5472..5975
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6012..6116
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6117..6185,6186..6977)
                     /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(6335..6354)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7148..7736
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7637..7656
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
       61 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
      121 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
      181 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
      241 caatttcaca caggaaacag ctatgaccat gattacgcca agctcgaaat taaccctcac
      301 taaagggaac aaaagctggt accgggcccc ccctcgaggt cgacaacacg ccgaataaac
      361 aaagggtttc ttcaatacct gatatagtag ctagagggca gaagtttgag gtgatatctt
      421 tctgagtgtt gttatttttg taagtcgtaa cacggaactg tttgcctgtt ggaattttac
      481 gttactattc tcatcggtat aactagctta tattcacttt ggcaataggt taaaaaaagc
      541 tcgttcattt ttttaaaaac agcaacgatg tcacacgaag gtgaagaaga tttattggag
      601 tactccgata acgaacaaga aattcaaatt ggtgcctcta aggccgctga agctggagaa
      661 actggcgccg ccacttctgc aactgaaggt gataataata acaacactgc agctggcgac
      721 aagaaaggtt cctatgttgg tatccattcc accggtttca aagatttctt gctaaagcca
      781 gaactatcaa gagccatcat tgactgtggt tttggacatc cttctgaggt ccagcaacat
      841 accattcctc agtcaattca tggtaccgat gtcttgtgtc aagcaaagtc tggtttaggt
      901 aagacagctg tctttgtctt atccactcta caacaattgg accctgttcc aggtgaagtt
      961 gccgttgttg tcatttgtaa tgctagagaa atggcctatc aaactcgtaa cgagtatttg
     1021 agattttcca aatatatgcc agacgttaag acagcagtct tttacggtgg tactccaatt
     1081 tctaaggatg ctgaactttt aaagaataaa gatactgctc ctcacattgt tgttgcaact
     1141 ccaggtcgtt taaaagcgtt agtgagagaa aaatacattg attcgtcaca cgtcaagaac
     1201 tttgtcatag atgaatgtga taaggtttta gaagaattag acatgagaag agatgtgcaa
     1261 gaaattttca gagctactcc aagagacaaa caagtcatga tgttttcagc cacactttct
     1321 caagaaatta gaccaatttg tagacgcttc ttacagaatc cattggaaat tttcgtcgat
     1381 gatgaagcta agctaacctt gcacgggttg caacaatatt atatcaaatt agaagaacgt
     1441 gaaaagaacc gtaaattggc tcaattatta gatgatttgg agttcaatca agtcattatt
     1501 ttcgttaaat ctaccacaag agctaatgag ttgaccaaac tattaaacgc ctctaacttt
     1561 ccggctatca ccgttcatgg tcatatgaaa caagaagaac gtattgctcg ttacaaggct
     1621 ttcaaagatt ttgaaaaacg tatttgcgtg tccacagatg tttttggtag aggtatcgat
     1681 attgaacgta ttaatttagc catcaattac gatttgacta atgaagctga ccaatattta
     1741 catcgtgtcg gtagagctgg tagatttggt actaagggtt tggctatttc attcgtttct
     1801 tcaaaggaag atgaggaagt cttggctaag atccaagagc gtttcgatgt caaaatcgct
     1861 gaattcccag aagaaggcat tgatccgtcc acttatttga ataattaata aaggaaaaaa
     1921 gatacgtttt tatatagatt atataaagat tttgtattat tcaaacgaac aaaacaaaca
     1981 aaaaagggga agtggaaaaa aattagcagt tatagcgaaa gattgagttt ttgttggaaa
     2041 atgaacgtag tgtagtcatt tcgtgaacgc acaagacaat tctattgggc atcacatggc
     2101 aaagcatgat aaggaaatga agttttggga atgattctaa attaacgttt tacccgttat
     2161 aggaagatat acagtatatg taagtaaaaa gttttttaat acaatagatg tgcaaagaaa
     2221 agtgaagaga tttctggaaa aaatcttgaa attttcttaa cggtaagatt tttggaatct
     2281 agagcggccc caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac
     2341 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2401 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2461 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2521 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2581 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2641 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2701 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2761 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2821 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2881 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     2941 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatc gaccggtcga
     3001 ggagaacttc tagtatatct acatacctaa tattattgcc ttattaaaaa tggaatccca
     3061 acaattacat caaaatccac attctcttca aaatcaattg tcctgtactt ccttgttcat
     3121 gtgtgttcaa aaacgttata tttataggat aattatactc tatttctcaa caagtaattg
     3181 gttgtttggc cgagcggtct aaggcgcctg attcaagaaa tatcttgacc gcagttaact
     3241 gtgggaatac tcaggtatcg taagatgcaa gagttcgaat ctcttagcaa ccattatttt
     3301 tttcctcaac ataacgagaa cacacagggg cgctatcgca cagaatcaaa ttcgatgact
     3361 ggaaattttt tgttaatttc agaggtcgcc tgacgcatat acctttttca actgaaaaat
     3421 tgggagaaaa aggaaaggtg agagcgccgg aaccggcttt tcatatagaa tagagaagcg
     3481 ttcatgacta aatgcttgca tcacaatact tgaagttgac aatattattt aaggacctat
     3541 tgttttttcc aataggtggt tagcaatcgt cttactttct aacttttctt accttttaca
     3601 tttcagcaat atatatatat atatttcaag gatataccat tctaatgtct gcccctaaga
     3661 agatcgtcgt tttgccaggt gaccacgttg gtcaagaaat cacagccgaa gccattaagg
     3721 ttcttaaagc tatttctgat gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa
     3781 ttggtggtgc tgctatcgat gctacaggtg ttccacttcc agatgaggcg ctggaagcct
     3841 ccaagaaggc tgatgccgtt ttgttaggtg ctgtgggtgg tcctaaatgg ggtaccggta
     3901 gtgttagacc tgaacaaggt ttactaaaaa tccgtaaaga acttcaattg tacgccaact
     3961 taagaccatg taactttgca tccgactctc ttttagactt atctccaatc aagccacaat
     4021 ttgctaaagg tactgacttc gttgttgtca gagaattagt gggaggtatt tactttggta
     4081 agagaaagga agacgatggt gatggtgtcg cttgggatag tgaacaatac accgttccag
     4141 aagtgcaaag aatcacaaga atggccgctt tcatggccct acaacatgag ccaccattgc
     4201 ctatttggtc cttggataaa gctaatgttt tggcctcttc aagattatgg agaaaaactg
     4261 tggaggaaac catcaagaac gaattcccta cattgaaggt tcaacatcaa ttgattgatt
     4321 ctgccgccat gatcctagtt aagaacccaa cccacctaaa tggtattata atcaccagca
     4381 acatgtttgg tgatatcatc tccgatgaag cctccgttat cccaggttcc ttgggtttgt
     4441 tgccatctgc gtccttggcc tctttgccag acaagaacac cgcatttggt ttgtacgaac
     4501 catgccacgg ttctgctcca gatttgccaa agaataaggt caaccctatc gccactatct
     4561 tgtctgctgc aatgatgttg aaattgtcat tgaacttgcc tgaagaaggt aaggccattg
     4621 aagatgcagt taaaaaggtt ttggatgcag gtatcagaac tggtgattta ggtggttcca
     4681 acagtaccac cgaagtcggt gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa
     4741 aagattctct ttttttatga tatttgtaca taaactttat aaatgaaatt cataatagaa
     4801 acgacacgaa attacaaaat ggaatatgtt catagggtag acgaaactat atacgcaatc
     4861 tacatacatt tatcaagaag gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt
     4921 gatactaatg gctcaacgtg ataaggaaaa agaattgcac tttaacatta atattgacaa
     4981 ggaggagggc accacacaaa aagttaggtg taacagaaaa tcatgaaact atgattccta
     5041 atttatatat tggaggattt tctctaaaaa aaaaaaaata caacaaataa aaaacactca
     5101 atgacctgac catttgatgg agtttaagtc aataccttct tgaaccattt cccataatgg
     5161 tgaaagttcc ctcaagaatt ttactctgtc agaaacggcc ttaacgacgt agtcgatatg
     5221 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     5281 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     5341 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     5401 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5461 ttcttaggac ggatcgcttg cctgtaactt acacgcgcct cgtatctttt aatgatggaa
     5521 taatttggga atttactctg tgtttattta tttttatgtt ttgtatttgg attttagaaa
     5581 gtaaataaag aaggtagaag agttacggaa tgaagaaaaa aaaataaaca aaggtttaaa
     5641 aaatttcaac aaaaagcgta ctttacatat atatttatta gacaagaaaa gcagattaaa
     5701 tagatataca ttcgattaac gataagtaaa atgtaaaatc acaggatttt cgtgtgtggt
     5761 cttctacaca gacaagatga aacaattcgg cattaatacc tgagagcagg aagagcaaga
     5821 taaaaggtag tatttgttgg cgatccccct agagtctttt acatcttcgg aaaacaaaaa
     5881 ctattttttc tttaatttct ttttttactt tctattttta atttatatat ttatattaaa
     5941 aaatttaaat tataattatt tttatagcac gtgatgaaaa ggacccaggt ggcacttttc
     6001 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6061 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6121 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6181 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6241 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6301 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6361 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6421 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6481 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6541 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6601 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6661 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6721 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6781 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6841 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6901 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6961 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7021 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7081 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7141 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7201 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7261 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7321 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7381 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7441 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7501 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7561 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7621 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7681 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7741 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     7801 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     7861 ccgctcgccg c
//
