LOCUS       sequence-133982-        7055 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Complementation of NME1 gene disruption or mutation in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7055)
  AUTHORS   Schmitt ME, Clayton DA
  TITLE     Nuclear RNase MRP is required for correct processing of pre-5.8S 
            rRNA in Saccharomyces cerevisiae.
  JOURNAL   Mol Cell Biol. 1993 Dec;13(12):7935-41.
  PUBMED    8247008
REFERENCE   2  (bases 1 to 7055)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Cell 
            Biol."; date: "1993-12"; volume: "13(12)"; pages: "7935-41"
COMMENT     Sequence Label: pMES140
FEATURES             Location/Qualifiers
     source          1..7055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(664..1758)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(1771..2175)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     rep_origin      complement(2475..2930)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2612..2633)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2824..2843
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3056..3078
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3070..3087
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3071..3087
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3094..3113
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3094..3112
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3145..3161
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3145..3161
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4236..4252)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4237..4253)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4281..4299)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4320..4336)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4320..4336)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4333..4355)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4344..4360
                     /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)."
     promoter        complement(join(4368..4374,4375..4392,4393..4398))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4413..4434
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4551..4568)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4722..5310)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5481..6272,6273..6341))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6104..6123
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6342..6446)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6483..6986
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     7028..7046
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatcga ctacgtcgtt aaggccgttt ctgacagagt aaaattcttg agggaacttt
      241 caccattatg ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc
      301 attgagtgtt ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatataa
      361 attaggaatc atagtttcat gattttctgt tacacctaac tttttgtgtg gtgccctcct
      421 ccttgtcaat attaatgtta aagtgcaatt ctttttcctt atcacgttga gccattagta
      481 tcaatttgct tacctgtatt cctttacatc ctcctttttc tccttcttga taaatgtatg
      541 tagattgcgt atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc
      601 gtttctatta tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc
      661 tttttaagca aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact
      721 gttggaacca cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc
      781 ttcaatggcc ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga
      841 caagatagtg gcgatagggt caaccttatt ctttggcaaa tctggagcag aaccgtggca
      901 tggttcgtac aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg
      961 caacaaaccc aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat
     1021 gttgctggtg attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc
     1081 agaatcaatc aattgatgtt gaaccttcaa tgtagggaat tcgttcttga tggtttcctc
     1141 cacagttttt ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat
     1201 aggcaatggt ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac
     1261 ttctggaacg gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct
     1321 cttaccaaag taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc
     1381 aaattgtggc ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct
     1441 taagttggcg tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac
     1501 actaccggta ccccatttag gaccacccac agcacctaac aaaacggcat caaccttctt
     1561 ggaggcttcc agcgcctcat ctggaagtgg gacacctgta gcatcgatag cagcaccacc
     1621 aattaaatga ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag
     1681 aaccttaatg gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat
     1741 cttcttaggg gcagacatag gggcagacat tagaatggta tatccttgaa atatatatat
     1801 atattgctga aatgtaaaag gtaagaaaag ttagaaagta agacgattgc taaccaccta
     1861 ttggaaaaaa caataggtcc ttaaataata ttgtcaactt caagtattgt gatgcaagca
     1921 tttagtcatg aacgcttctc tattctatat gaaaagccgg ttccggcctc tcacctttcc
     1981 tttttctccc aatttttcag ttgaaaaagg tatatgcgtc aggcgacctc tgaaattaac
     2041 aaaaaatttc cagtcatcga atttgattct gtgcgatagc gcccctgtgt gttctcgtta
     2101 tgttgaggaa aaaaataatg gttgctaaga gattcgaact cttgcatctt acgatacctg
     2161 agtattccca cagttaactg cggtcaagat atttcttgaa tcaggcgcct tagaccgctc
     2221 ggccaaacaa ccaattactt gttgagaaat agagtataat tatcctataa atataacgtt
     2281 tttgaacaca catgaacaag gaagtacagg acaattgatt ttgaagagaa tgtggatttt
     2341 gatgtaattg ttgggattcc atttttaata aggcaataat attaggtatg tggatatact
     2401 agaagttctc ctcgagggtc gatatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     2461 aataccgcat caggaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     2521 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     2581 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     2641 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     2701 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     2761 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     2821 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     2881 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt ccattcgcca
     2941 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     3001 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     3061 tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg
     3121 gagctccacc gcggtggcgg ccgctctaga actagtggat cccccgggct gcaggaattc
     3181 ctcgagctac gtcagggctc ttcaactaaa gtttgagagg gcattttttg ttcgaacaat
     3241 atcagtgcta acaaggttgc taccacatca atgaactcta ctcctgtatg aaccctaatt
     3301 ttccttcatg aaaaatgaat tttcgatgtt tttccgtgga ttttatattt tttcaaatat
     3361 ataaaggagt aagaaaataa ctttgtttat cctatgttgc ggtaaagaac taaatagtgg
     3421 tcttcacctg ttttctaatt tcttctcgtt taaagttctt caatccatga ccaaagaatc
     3481 gtcacaaatc gaagcttaca aaatggagta aaattttttt tactcagtaa tatgctttgg
     3541 gttgaaagtc tcccaccaat tcgtatgcgg aaaacgtaat gagatttaaa aattttaaat
     3601 tgtttaaatc aactcattaa ggaggatgcc cttgggtatt ctgcttcttg acctggtacc
     3661 tctattgcag ggtactggtg ttttcttcgg tactggattc cgtttgtatg gaatctaaac
     3721 catagttatg acgattgctc tttcccgtgc tggatcgagt aacccaatgg agcttactat
     3781 tcttggtcca tggattcacc cctttttatt tttttaacag tttgttgttt acactatttt
     3841 ttagcagtgt aaaacttttg catagttaaa tatgccgaac tgatctaata aaaagtctga
     3901 ataactcaga tgagttattt attggtaaac ttgtgtaaga agaagatgga atagtgaata
     3961 attcatcaaa tgatgaaata ccaatgcaac agagtcaagc tctgagtttc aaaaagaaac
     4021 atggacgaga ttgctttttt attactgacc atgattattc attacttacc attgttcctc
     4081 attctatata attgattggg caccgcgtaa attcttgagc ctcttcggta gtattcaatc
     4141 gcaatatggg gtagtttacg gaagtgctgt aaatgatgtc taaaatgaac aactatgcgt
     4201 ccttattgtg acaatgaatt cgatatcaag cttatcgata ccgtcgacct cgaggggggg
     4261 cccggtacca gcttttgttc cctttagtga gggttaattt cgagcttggc gtaatcatgg
     4321 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     4381 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     4441 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4501 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4561 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4621 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4681 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4741 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4801 taaagatacc aggcgttccc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4861 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     4921 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4981 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5041 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5101 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5161 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5221 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5281 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5341 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5401 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5461 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5521 tgtctatttc gttcatccat agttgcctga ctgcccgtcg tgtagataac tacgatacgg
     5581 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5641 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5701 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5761 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5821 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5881 cccatgttgt gaaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5941 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6001 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6061 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6121 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6181 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6241 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6301 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     6361 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     6421 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgggtccttt
     6481 tcatcacgtg ctataaaaat aattataatt taaatttttt aatataaata tataaattaa
     6541 aaatagaaag taaaaaaaga aattaaagaa aaaatagttt ttgttttccg aagatgtaaa
     6601 agactctagg gggatcgcca acaaatacta ccttttatct tgctcttcct gctctcaggt
     6661 attaatgccg aattgtttca tcttgtctgt gtagaagacc acacacgaaa atcctgtgat
     6721 tttacatttt acttatcgtt aatcgaatgt atatctattt aatctgcttt tcttgtctaa
     6781 taaatatata tgtaaagtac gctttttgtt gaaatttttt aaacctttgt ttattttttt
     6841 ttcttcattc cgtaactctt ctaccttctt tatttacttt ctaaaatcca aatacaaaac
     6901 ataaaaataa ataaacacag agtaaattcc caaattattc catcattaaa agatacgagg
     6961 cgcgtgtaag ttacaggcaa gcgatccgtc ctaagaaacc attattatca tgacattaac
     7021 ctataaaaat aggcgtatca cgaggccctt tcgtc
//
