LOCUS       sequence-178972-        6669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6669)
  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 6669)
  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: Sub2-3 in pRS314
FEATURES             Location/Qualifiers
     source          1..6669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1748..1767)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1749..1767)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1774..1791)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1774..1790)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1783..1805)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1931..2386
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2018..2037)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2228..2249
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2489..3163)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3164..3444)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3465..3484)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3584..3606
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3644..3662)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3703..4206
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4243..4347
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4348..4416,4417..5208)
                     /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(4566..4585)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5379..5967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5868..5887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6121..6138
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6255..6276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6291..6296,6297..6314,6315..6321)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6329..6345
                     /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     6334..6356
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6353..6369
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6353..6369
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        6390..6408
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 atgtcacacg aaggtgaaga agatttattg gagtactccg ataacgaaca agaaattcaa
       61 attgatgcct ctaaggccgc tgaagctgga gaaactggcg ccgccacttc tgcaactgaa
      121 ggtgataata ataacaacac tgcagctggc gacaagaaag gttcctatgt tggtatccat
      181 tccaccggtt tcaaagattt cttgctaaag ccagaactat caagagccat cattgactgt
      241 ggttttgaac atccttctga ggtccagcaa cataccattc ctcagtcaat tcatggtacc
      301 gatgtcttgt gtcaagcaaa gtctggttta ggtaagacag ctgtctttgt cttatccact
      361 ctgcaacaat tggaccctgt tccaggtgaa gttgccgttg ttgtcatttg taatgctaga
      421 gaactggcct atcaaattcg taacgagtat ttgagatttt ccaaatatat gccagacgtt
      481 aagacagcag tcttatacgg tggtactcca atttctaagg atgctgaact tttaaagaat
      541 aaagatactg ctcctcacat tgttgttgca actccaggtc gtttaaaagc gttagtgaga
      601 gaaaaataca ttgatttgtc acacgtcaag aactttgtca tagatgaatg tgataaggtt
      661 ttagaagaat tagacatgag aagagatgtg caagaaattt tcagagctac tccaagagac
      721 aaacaagtca tgatgttttc agctacactt tctcaagaaa ttagaccaat ttgtagacgc
      781 ttcttacaga atccattgga aattttcgtc gacgatgaag ctaagctaac cttgcacggg
      841 ttgcaacaat attatatcaa attagaagaa cgtgaaaaga accgtaaatt ggctcaatta
      901 ttagatgatt tggagttcaa tcaagtcatt attttcgtta aatctaccac aagagctaat
      961 gagttgacca aactattaaa cgcctctaac tttccggcta tcaccgttca tggtcatatg
     1021 aaacaagaag aacgtattgc tcgttacaag gctttcaaag attttgaaaa acgtatttgc
     1081 gtgtccacag atgtttttgg tagaggtatc gatattgaac gtattaattt agccatcaat
     1141 tacgatttga ctaatgaagc tgaccaattt ttacatcgtg tcggtagagc tggtagattt
     1201 ggtactaagg gtttggctat ttcattcgtt tcttcaaagg aagatgagga agtcttggct
     1261 aagatccaag agcgtttcga tgtctaaatc gctgaattcc cagaagaagg cattgatccg
     1321 tccacttatt tgaataatta ataaaggaaa aaagatacgt ttttatatag attatataaa
     1381 gattttgtat tattcagacg aacaaaacaa acaaaaaagg ggaagtggaa aaaaattagc
     1441 agttatagcg aaagattgag tttttgttgg aaaatgaacg tagtgtagtc atttcgtgaa
     1501 cgcacaagac aattctattg ggcatctcat ggcaaagcat gataaggaaa tgaagttttg
     1561 ggaatgattc taaattaacg ttttacccgt tataggaaga tatacagtat atgtaagtaa
     1621 aaagtttttt aatacaatag atgtgcaaag aaaagtgaag agatttctgg aaaaaatctt
     1681 gaaattttct taacggtaag atttttggaa tctagagcgg ccgccaccgc ggtggagctc
     1741 caattcgccc tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga
     1801 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     1861 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     1921 tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1981 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2041 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2101 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2161 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2221 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2281 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2341 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt
     2401 ttctccttac gcatctgtgc ggtatttcac accgcataga tcggcaagtg cacaaacaat
     2461 acttaaataa atactactca gtaataacct atttcttagc atttttgacg aaatttgcta
     2521 ttttgttaga gtcttttaca ccatttgtct ccacacctcc gcttacatca acaccaataa
     2581 cgccatttaa tctaagcgca tcaccaacat tttctggcgt cagtccacca gctaacataa
     2641 aatgtaagct ttcggggctc tcttgccttc caacccagtc agaaatcgag ttccaatcca
     2701 aaagttcacc tgtcccacct gcttctgaat caaacaaggg aataaacgaa tgaggtttct
     2761 gtgaagctgc actgagtagt atgttgcagt cttttggaaa tacgagtctt ttaataactg
     2821 gcaaaccgag gaactcttgg tattcttgcc acgactcatc tccatgcagt tggacgatat
     2881 caatgccgta atcattgacc agagccaaaa catcctcctt aggttgatta cgaaacacgc
     2941 caaccaagta tttcggagtg cctgaactat ttttatatgc ttttacaaga cttgaaattt
     3001 tccttgcaat aaccgggtca attgttctct ttctattggg cacacatata atacccagca
     3061 agtcagcatc ggaatctaga gcacattctg cggcctctgt gctctgcaag ccgcaaactt
     3121 tcaccaatgg accagaacta cctgtgaaat taataacaga catactccaa gctgcctttg
     3181 tgtgcttaat cacgtatact cacgtgctca atagtcacca atgccctccc tcttggccct
     3241 ctccttttct tttttcgacc gaattaattc ttaatcggca aaaaaagaaa agctccggat
     3301 caagattgta cgtaaggtga caagctattt ttcaataaag aatatcttcc actactgcca
     3361 tctggcgtca taactgcaaa gtacacatat attacgatgc tgttctatta aatgcttcct
     3421 atattatata tatagtaatg tcgtgatcta tggtgcactc tcagtacaat ctgctctgat
     3481 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     3541 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     3601 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     3661 tttttatagg ttaatgtcat gataataatg gtttcttaga cggatcgctt gcctgtaact
     3721 tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct gtgtttattt
     3781 atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa gagttacgga
     3841 atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt actttacata
     3901 tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa cgataagtaa
     3961 aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg aaacaattcg
     4021 gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg gcgatccccc
     4081 tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc tttttttact
     4141 ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat ttttatagca
     4201 cgtgatgaaa aggacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     4261 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     4321 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     4381 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     4441 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     4501 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     4561 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     4621 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     4681 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     4741 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     4801 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     4861 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     4921 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     4981 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     5041 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     5101 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     5161 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     5221 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5281 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5341 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5401 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5461 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5521 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5581 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5641 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5701 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5761 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5821 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5881 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5941 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6001 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6061 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6121 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg
     6181 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
     6241 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
     6301 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
     6361 agctatgacc atgattacgc caagctcgaa attaaccctc actaaaggga acaaaagctg
     6421 gtaccgggcc ccccctcgag gtcgacaaca cgccgaataa acaaagggtt tcttcaatac
     6481 ctgatatagt agctagaggg cagaagtttg aggtgatatc tttctgagtg ttgttatttt
     6541 tgtaagtcgt aacacggaac tgtttgcctg ttggaatttt acgttactat tctcatcggt
     6601 ataactagct tatattcact ttggcaatag gttaaaaaaa gctcgttcat ttttttaaaa
     6661 acagcaacg
//
