LOCUS       sequence-217249-        7871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sub2 in pRS315 vector.
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 pRS315
FEATURES             Location/Qualifiers
     source          1..7871
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..24
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    141..162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(177..182,183..200,201..207)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    215..231
                     /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     220..242
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     239..255
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     239..255
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        276..294
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2286..2305)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2287..2305)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2312..2329)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2312..2328)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2321..2343)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2469..2924
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2556..2575)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2766..2787
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3225..3632
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3633..4727
                     /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(5221..5240)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5340..5362
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5400..5418)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5460..5963
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6000..6104
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6105..6173,6174..6965)
                     /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(6323..6342)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7136..7724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7625..7644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
       61 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
      121 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
      181 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
      241 ggaaacagct atgaccatga ttacgccaag ctcgaaatta accctcacta aagggaacaa
      301 aagctggtac cgggcccccc ctcgaggtcg acaacacgcc gaataaacaa agggtttctt
      361 caatacctga tatagtagct agagggcaga agtttgaggt gatatctttc tgagtgttgt
      421 tatttttgta agtcgtaaca cggaactgtt tgcctgttgg aattttacgt tactattctc
      481 atcggtataa ctagcttata ttcactttgg caataggtta aaaaaagctc gttcattttt
      541 ttaaaaacag caacgatgtc acacgaaggt gaagaagatt tattggagta ctccgataac
      601 gaacaagaaa ttcaaattga tgcctctaag gccgctgaag ctggagaaac tggcgccgcc
      661 acttctgcaa ctgaaggtga taataataac aacactgcag ctggcgacaa gaaaggttcc
      721 tatgttggta tccattccac cggtttcaaa gatttcttgc taaagccaga actatcaaga
      781 gccatcattg actgtggttt tgaacatcct tctgaggtcc agcaacatac cattcctcag
      841 tcaattcatg gtaccgatgt cttgtgtcaa gcaaagtctg gtttaggtaa gacagctgtc
      901 tttgtcttat ccactctgca acaattggac cctgttccag gtgaagttgc cgttgttgtc
      961 atttgtaatg ctagagaact ggcctatcaa attcgtaacg agtatttgag attttccaaa
     1021 tatatgccag acgttaagac agcagtcttt tacggtggta ctccaatttc taaggatgct
     1081 gaacttttaa agaataaaga tactgctcct cacattgttg ttgcaactcc aggtcgttta
     1141 aaagcgttag tgagagaaaa atacattgat ttgtcacacg tcaagaactt tgtcatagat
     1201 gaatgtgata aggttttaga agaattagac atgagaagag atgtgcaaga aattttcaga
     1261 gctactccaa gagacaaaca agtcatgatg ttttcagcca cactttctca agaaattaga
     1321 ccaatttgta gacgcttctt acagaatcca ttggaaattt tcgtcgatga tgaagctaag
     1381 ctaaccttgc acgggttgca acaatattat atcaaattag aagaacgtga aaagaaccgt
     1441 aaattggctc aattattaga tgatttggag ttcaatcaag tcattatttt cgttaaatct
     1501 accacaagag ctaatgagtt gaccaaacta ttaaacgcct ctaactttcc ggctatcacc
     1561 gttcatggtc atatgaaaca agaagaacgt attgctcgtt acaaggcttt caaagatttt
     1621 gaaaaacgta tttgcgtgtc cacagatgtt tttggtagag gtatcgatat tgaacgtatt
     1681 aatttagcca tcaattacga tttgactaat gaagctgacc aatatttaca tcgtgtcggt
     1741 agagctggta gatttggtac taagggtttg gctatttcat tcgtttcttc aaaggaagat
     1801 gaggaagtct tggctaagat ccaagagcgt ttcgatgtca aaatcgctga attcccagaa
     1861 gaaggcattg atccgtccac ttatttgaat aattaataaa ggaaaaaaga tacgttttta
     1921 tatagattat ataaagattt tgtattattc aaacgaacaa aacaaacaaa aaaggggaag
     1981 tggaaaaaaa ttagcagtta tagcgaaaga ttgagttttt gttggaaaat gaacgtagtg
     2041 tagtcatttc gtgaacgcac aagacaattc tattgggcat cacatggcaa agcatgataa
     2101 ggaaatgaag ttttgggaat gattctaaat taacgtttta cccgttatag gaagatatac
     2161 agtatatgta agtaaaaagt tttttaatac aatagatgtg caaagaaaag tgaagagatt
     2221 tctggaaaaa atcttgaaat tttcttaacg gtaagatttt tggaatctag agcggcccca
     2281 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     2341 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2401 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2461 gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2521 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2581 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2641 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2701 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2761 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2821 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2881 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     2941 ctccttacgc atctgtgcgg tatttcacac cgcatatcga ccggtcgagg agaacttcta
     3001 gtatatctac atacctaata ttattgcctt attaaaaatg gaatcccaac aattacatca
     3061 aaatccacat tctcttcaaa atcaattgtc ctgtacttcc ttgttcatgt gtgttcaaaa
     3121 acgttatatt tataggataa ttatactcta tttctcaaca agtaattggt tgtttggccg
     3181 agcggtctaa ggcgcctgat tcaagaaata tcttgaccgc agttaactgt gggaatactc
     3241 aggtatcgta agatgcaaga gttcgaatct cttagcaacc attatttttt tcctcaacat
     3301 aacgagaaca cacaggggcg ctatcgcaca gaatcaaatt cgatgactgg aaattttttg
     3361 ttaatttcag aggtcgcctg acgcatatac ctttttcaac tgaaaaattg ggagaaaaag
     3421 gaaaggtgag agcgccggaa ccggcttttc atatagaata gagaagcgtt catgactaaa
     3481 tgcttgcatc acaatacttg aagttgacaa tattatttaa ggacctattg ttttttccaa
     3541 taggtggtta gcaatcgtct tactttctaa cttttcttac cttttacatt tcagcaatat
     3601 atatatatat atttcaagga tataccattc taatgtctgc ccctaagaag atcgtcgttt
     3661 tgccaggtga ccacgttggt caagaaatca cagccgaagc cattaaggtt cttaaagcta
     3721 tttctgatgt tcgttccaat gtcaagttcg atttcgaaaa tcatttaatt ggtggtgctg
     3781 ctatcgatgc tacaggtgtt ccacttccag atgaggcgct ggaagcctcc aagaaggctg
     3841 atgccgtttt gttaggtgct gtgggtggtc ctaaatgggg taccggtagt gttagacctg
     3901 aacaaggttt actaaaaatc cgtaaagaac ttcaattgta cgccaactta agaccatgta
     3961 actttgcatc cgactctctt ttagacttat ctccaatcaa gccacaattt gctaaaggta
     4021 ctgacttcgt tgttgtcaga gaattagtgg gaggtattta ctttggtaag agaaaggaag
     4081 acgatggtga tggtgtcgct tgggatagtg aacaatacac cgttccagaa gtgcaaagaa
     4141 tcacaagaat ggccgctttc atggccctac aacatgagcc accattgcct atttggtcct
     4201 tggataaagc taatgttttg gcctcttcaa gattatggag aaaaactgtg gaggaaacca
     4261 tcaagaacga attccctaca ttgaaggttc aacatcaatt gattgattct gccgccatga
     4321 tcctagttaa gaacccaacc cacctaaatg gtattataat caccagcaac atgtttggtg
     4381 atatcatctc cgatgaagcc tccgttatcc caggttcctt gggtttgttg ccatctgcgt
     4441 ccttggcctc tttgccagac aagaacaccg catttggttt gtacgaacca tgccacggtt
     4501 ctgctccaga tttgccaaag aataaggtca accctatcgc cactatcttg tctgctgcaa
     4561 tgatgttgaa attgtcattg aacttgcctg aagaaggtaa ggccattgaa gatgcagtta
     4621 aaaaggtttt ggatgcaggt atcagaactg gtgatttagg tggttccaac agtaccaccg
     4681 aagtcggtga tgctgtcgcc gaagaagtta agaaaatcct tgcttaaaaa gattctcttt
     4741 ttttatgata tttgtacata aactttataa atgaaattca taatagaaac gacacgaaat
     4801 tacaaaatgg aatatgttca tagggtagac gaaactatat acgcaatcta catacattta
     4861 tcaagaagga gaaaaaggag gatgtaaagg aatacaggta agcaaattga tactaatggc
     4921 tcaacgtgat aaggaaaaag aattgcactt taacattaat attgacaagg aggagggcac
     4981 cacacaaaaa gttaggtgta acagaaaatc atgaaactat gattcctaat ttatatattg
     5041 gaggattttc tctaaaaaaa aaaaaataca acaaataaaa aacactcaat gacctgacca
     5101 tttgatggag tttaagtcaa taccttcttg aaccatttcc cataatggtg aaagttccct
     5161 caagaatttt actctgtcag aaacggcctt aacgacgtag tcgatatggt gcactctcag
     5221 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5281 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5341 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5401 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttaggacgg
     5461 atcgcttgcc tgtaacttac acgcgcctcg tatcttttaa tgatggaata atttgggaat
     5521 ttactctgtg tttatttatt tttatgtttt gtatttggat tttagaaagt aaataaagaa
     5581 ggtagaagag ttacggaatg aagaaaaaaa aataaacaaa ggtttaaaaa atttcaacaa
     5641 aaagcgtact ttacatatat atttattaga caagaaaagc agattaaata gatatacatt
     5701 cgattaacga taagtaaaat gtaaaatcac aggattttcg tgtgtggtct tctacacaga
     5761 caagatgaaa caattcggca ttaatacctg agagcaggaa gagcaagata aaaggtagta
     5821 tttgttggcg atccccctag agtcttttac atcttcggaa aacaaaaact attttttctt
     5881 taatttcttt ttttactttc tatttttaat ttatatattt atattaaaaa atttaaatta
     5941 taattatttt tatagcacgt gatgaaaagg acccaggtgg cacttttcgg ggaaatgtgc
     6001 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6061 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6121 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6181 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6241 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6301 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     6361 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6421 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6481 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     6541 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6601 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     6661 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6721 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6781 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6841 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6901 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6961 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7021 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7081 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7141 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7201 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7261 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7321 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7381 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7441 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7501 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7561 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7621 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7681 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7741 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7801 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     7861 gccgaacgac c
//
