LOCUS       sequence-405869-        8265 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression vector for hNUS1 R290H.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8265)
  AUTHORS   Park EJ, Grabinska KA, Guan Z, Stranecky V, Hartmannova H, Hodanova 
            K, Baresova V, Sovova J, Jozsef L, Ondruskova N, Hansikova H, Honzik
            T, Zeman J, Hulkova H, Wen R, Kmoch S, Sessa WC
  TITLE     Mutation of Nogo-B receptor, a subunit of cis-prenyltransferase, 
            causes a congenital disorder of glycosylation.
  JOURNAL   Cell Metab. 2014 Sep 2;20(3):448-57. doi: 
            10.1016/j.cmet.2014.06.016. Epub 2014 Jul 24.
  PUBMED    25066056
REFERENCE   2  (bases 1 to 8265)
  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"; doi: 
            "10.1016/j.cmet.2014.06.016"; journalName: "Cell Metab"; date: 
            "2014-09-2- 2"; volume: "20"; issue: "3"; pages: "448-57"
COMMENT     Sequence Label: pKG-GW2-NgBR R290H
FEATURES             Location/Qualifiers
     source          1..8265
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        43..709
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     579..602
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     protein_bind    712..736
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1658..1682)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1887..1904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2058..2646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2138..2157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2817..3608,3609..3677))
                     /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     3440..3459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3678..3782)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3850..3868
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3906..3928)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4028..4047
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4064..5046
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5047..5452
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        5742..6091
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             6092..7426
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHSTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENARKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
ORIGIN
        1 aatcattttg aataaaaaac acgctttttc agttcgagtt tatcattatc aatactgcca
       61 tttcaaagaa tacgtaaata attaatagta gtgattttcc taactttatt tagtcaaaaa
      121 attagccttt taattctgct gtaacccgta catgcccaaa atagggggcg ggttacacag
      181 aatatataac atcgtaggtg tctgggtgaa cagtttattc ctggcatcca ctaaatataa
      241 tggagcccgc tttttaagct ggcatccaga aaaaaaaaga atcccagcac caaaatattg
      301 ttttcttcac caaccatcag ttcataggtc cattctctta gcgcaactac agagaacagg
      361 ggcacaaaca ggcaaaaaac gggcacaacc tcaatggagt gatgcaacct gcctggagta
      421 aatgatgaca caaggcaatt gacccacgca tgtatctatc tcattttctt acaccttcta
      481 ttaccttctg ctctctctga tttggaaaaa gctgaaaaaa aaggttgaaa ccagttccct
      541 gaaattattc ccctacttga ctaataagta tataaagacg gtaggtattg attgtaattc
      601 tgtaaatcta tttcttaaac ttcttaaatt ctacttttat agttagtctt ttttttagtt
      661 ttaaaacacc aagaacttag tttcgaataa acacacataa acagaattat cacaagtttg
      721 tacaaaaaag caggctccga attcgccctt ggatccatga cggggctgta cgagctggtg
      781 tggcgggtgc tgcacgcgct gctctgtctg caccgcacgc tcacctcctg gctccgcgtt
      841 cggttcggca cctggaactg gatctggcgg cgctgctgcc gcgccgcctc tgccgcggtc
      901 ctagcgccgc tcggcttcac gctccgcaag cccccggcag tcggcaggaa ccgccgtcac
      961 caccggcacc cgcgcggggg gtcgtgcctg gcagccgcac accaccggat gcgctggcgc
     1021 gcggacggtc gttccttgga gaagctgcct gtgcatatgg gcctggtgat caccgaggtg
     1081 gagcaggaac ccagcttctc ggacatcgcg agcctcgtgg tgtggtgtat ggccgtgggc
     1141 atctcctaca ttagcgtcta cgaccaccaa ggtattttca aaagaaataa ttccagattg
     1201 atggatgaaa ttttaaaaca acagcaagaa cttctgggcc tagattgttc aaaatactca
     1261 ccagaatttg caaatagtaa tgacaaagat gatcaagttt taaattgcca tttggcagtg
     1321 aaggtgctgt ctccggaaga tggaaaagca gatattgtaa gagctgctca ggacttttgc
     1381 cagttagtag cccagaagca aaagagaccc acagatttgg atgtagatac gttagccagt
     1441 ttacttagtt caaatggttg tcctgatcct gatttagtat tgaagttcgg tcctgtggac
     1501 agcacattag gctttcttcc ctggcacatc agattgactg agattgtctc tttgccttcc
     1561 cacctaaaca tcagttatga ggactttttc tctgcccttc gtcaatatgc agcctgtgaa
     1621 cagcatctgg gaaagtaggt cgacaagggc gaattcgacc cagctttctt gtacaaagtg
     1681 gtgattcgac ttggttgaac acgttgccaa ggcttaagtg aatttacttt aaatcttgca
     1741 tttaaataaa ttttcttttt atagctttat gacttagttt caatttatat actattttaa
     1801 tgacattttc gattcggatc cgtgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     1861 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     1921 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     1981 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     2041 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     2101 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2161 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     2221 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     2281 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     2341 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     2401 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     2461 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     2521 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     2581 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     2641 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     2701 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     2761 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     2821 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     2881 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     2941 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3001 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3061 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3121 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3181 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3241 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3301 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3361 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3421 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3481 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3541 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3601 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3661 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     3721 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     3781 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     3841 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     3901 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     3961 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     4021 aactatgcgg catcagagca gattgtactg agagtgcacc atactagaaa gtataggaac
     4081 ttcagagcgc ttttgaaaac caaaagcgct ctgaagacgc actttcaaaa aaccaaaaac
     4141 gcaccggact gtaacgagct actaaaatat tgcgaatacc gcttccacaa acattgctca
     4201 aaagtatctc tttgctatat atctctgtgc tatatcccta tataacctac ccatccacct
     4261 ttcgctcctt gaacttgcat ctaaactcga cctctacatt ttttatgttt atctctagta
     4321 ttactcttta gacaaaaaaa ttgtagtaag aactattcat agagtgaatc gaaaacaata
     4381 cgaaaatgta aacatttcct atacgtagta tatagagaca aaatagaaga aaccgttcat
     4441 aattttctga ccaatgaaga atcatcaacg ctatcacttt ctgttcacaa agtatgcgca
     4501 atccacatcg gtatagaata taatcgggga tgcctttatc ttgaaaaaat gcacccgcag
     4561 cttcgctagt aatcagtaaa cgcgggaagt ggagtcaggc tttttttatg gaagagaaaa
     4621 tagacaccaa agtagccttc ttctaacctt aacggaccta cagtgcaaaa agttatcaag
     4681 agactgcatt atagagcgca caaaggagaa aaaaagtaat ctaagatgct ttgttagaaa
     4741 aatagcgctc tcgggatgca tttttgtaga acaaaaaaga agtatagatt ctttgttggt
     4801 aaaatagcgc tctcgcgttg catttctgtt ctgtaaaaat gcagctcaga ttctttgttt
     4861 gaaaaattag cgctctcgcg ttgcattttt gttttacaaa aatgaagcac agattcttcg
     4921 ttggtaaaat agcgctttcg cgttgcattt ctgttctgta aaaatgcagc tcagattctt
     4981 tgtttgaaaa attagcgctc tcgcgttgca tttttgttct acaaaatgaa gcacagatgc
     5041 ttcgttaact gtgggaatac tcaggtatcg taagatgcaa gagttcgaat ctcttagcaa
     5101 ccattatttt tttcctcaac ataacgagaa cacacagggg cgctatcgca cagaatcaaa
     5161 ttcgatgact ggaaattttt tgttaatttc agaggtcgcc tgacgcatat acctttttca
     5221 actgaaaaat tgggagaaaa aggaaaggtg agagcgccgg aaccggcttt tcatatagaa
     5281 tagagaagcg ttcatgacta aatgcttgca tcacaatact tgaagttgac aatattattt
     5341 aaggacctat tgttttttcc aataggtggt tagcaatcgt cttactttct aacttttctt
     5401 accttttaca tttcagcaat atatatatat atttcaagga tataccattc taatgtctgc
     5461 ccctaagaag atcgtcgttt tgccaggtga ccataatata ctagtaacgt aaatactagt
     5521 tagtagatga tagttgattt ttattccaac actaagaaat aatttcgcca tttcttgaat
     5581 gtatttaaag atatttaatg ctataataga catttaaatc caattcttcc aacatacaat
     5641 gggagtttgg ccgagtggtt taaggcgtca gatttaggtg gatttaacct ctaaaatctc
     5701 tgatatcttc ggatgcaagg gttcgaatcc cttagctctc attatttttt gctttttctc
     5761 ttgaggtcac atgatcgcaa aatggcaaat ggcacgtgaa gctgtcgata ttggggaact
     5821 gtggtggttg gcaaatgact aattaagtta gtcaaggcgc catcctcatg aaaactgtgt
     5881 aacataataa ccgaagtgtc gaaaaggtgg caccttgtcc aattgaacac gctcgatgaa
     5941 aaaataagat atatataagg ttaagtaaag cgtctgttag aaaggaagtt tttccttttt
     6001 cttgctctct tgtcttttca tctactattt ccttcgtgta atacagggtc gtcagataca
     6061 tagatacaat tctattaccc ccatccatac aatgccatct catttcgata ctgttcaact
     6121 acacgccggc caagagaacc ctggtgacaa tgctcacaga tccagagctg taccaattta
     6181 cgccaccact tcttatgttt tcgaaaactc taagcatggt tcgcaattgt ttggtctaga
     6241 agttccaggt tacgtctatt cccgtttcca aaacccaacc agtaatgttt tggaagaaag
     6301 aattgctgct ttagaaggtg gtgctgctgc tttggctgtt tcctccggtc aagccgctca
     6361 aacccttgcc atccaaggtt tggcacacac tggtgacaac atcgtttcca cttcttactt
     6421 atacggtggt acttataacc agttcaaaat ctcgttcaaa agatttggta tcgaggctag
     6481 atttgttgaa ggtgacaatc cagaagaatt cgaaaaggtc tttgatgaaa gaaccaaggc
     6541 tgtttatttg gaaaccattg gtaatccaaa gtacaatgtt ccggattttg aaaaaattgt
     6601 tgcaattgct cacaaacacg gtattccagt tgtcgttgac aacacatttg gtgccggtgg
     6661 ttacttctgt cagccaatta aatacggtgc tgatattgta acacattctg ctaccaaatg
     6721 gattggtggt catggtacta ctatcggtgg tattattgtt gactctggta agttcccatg
     6781 gaaggactac ccagaaaagt tccctcaatt ctctcaacct gccgaaggat atcacagtac
     6841 tatctacaat gaagcctacg gtaacttggc atacatcgtt catgttagaa ctgaactatt
     6901 aagagatttg ggtccattga tgaacccatt tgcctctttc ttgctactac aaggtgttga
     6961 aacattatct ttgagagctg aaagacacgg tgaaaatgca ttgaagttag ccaaatggtt
     7021 agaacaatcc ccatacgtat cttgggtttc ataccctggt ttagcatctc attctcatca
     7081 tgaaaatgct aggaagtatc tatctaacgg tttcggtggt gtcttatctt tcggtgtaaa
     7141 agacttacca aatgccgaca aggaaactga cccattcaaa ctttctggtg ctcaagttgt
     7201 tgacaattta aagcttgcct ctaacttggc caatgttggt gatgccaaga ccttagtcat
     7261 tgctccatac ttcactaccc acaaacaatt aaatgacaaa gaaaagttgg catctggtgt
     7321 taccaaggac ttaattcgtg tctctgttgg tatcgaattt attgatgaca ttattgcaga
     7381 cttccagcaa tcttttgaaa ctgttttcgc tggccaaaaa ccatgagtgt gcgtaatgag
     7441 ttgtaaaatt atgtataaac ctactttctc tcacaagtac tatactttta taaaacgaac
     7501 tttattgaaa tgaatatcct ttttttccct tgttacatgt cgtgactcgt actttgaacc
     7561 taaattgttc taacatcaaa gaacagtgtt aattcgcagt cgagaagaaa aatatggtga
     7621 acaagactca tctacttcat gagactactt tacgcctcct ataaagctgt cacactgtac
     7681 ataaacttta taaatgaaat tcataataga aacgacacga aattacaaaa tggaatatgt
     7741 tcatagggta gacgaaacta tatacgcaat ctacatacat ttatcaagaa ggagaaaaag
     7801 gaggatgtaa aggaatacag gtaagcaaat tgatactaat ggctcaacgt gataaggaaa
     7861 aagaattgca ctttaacatt aatattgaca aggaggaggg caccacacaa aaagttaggt
     7921 gtaacagaaa atcatgaaac tatgattcct aatttatata ttggaggatt ttctctaaaa
     7981 aaaaaaaaat acaacaaata aaaaacactc aatgacctga ccatttgatg gagtttaagt
     8041 caataccttc ttgaaccatt tcccataatg gtgaaagttc cctcaagaat tttactctgt
     8101 cagaaacggc cttaacgacg taccggatct atgcggtgtg aaataccgca cagatgcgta
     8161 aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg
     8221 cgatcggtgc gggcctcttc gctattacgc cagcggatcc ggtag
//
