LOCUS       sequence-400037-        8267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression vector for hNUS1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8267)
  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 8267)
  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
FEATURES             Location/Qualifiers
     source          1..8267
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        54..720
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     590..613
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     protein_bind    723..747
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1671..1695)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1900..1917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2071..2659)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2151..2170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2830..3621,3622..3690))
                     /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     3453..3472
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3691..3795)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3863..3881
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3919..3941)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4041..4060
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4077..5059
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5060..5465
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        5755..6104
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             6105..7439
                     /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 ggatccggta gaatcatttt gaataaaaaa cacgcttttt cagttcgagt ttatcattat
       61 caatactgcc atttcaaaga atacgtaaat aattaatagt agtgattttc ctaactttat
      121 ttagtcaaaa aattagcctt ttaattctgc tgtaacccgt acatgcccaa aatagggggc
      181 gggttacaca gaatatataa catcgtaggt gtctgggtga acagtttatt cctggcatcc
      241 actaaatata atggagcccg ctttttaagc tggcatccag aaaaaaaaag aatcccagca
      301 ccaaaatatt gttttcttca ccaaccatca gttcataggt ccattctctt agcgcaacta
      361 cagagaacag gggcacaaac aggcaaaaaa cgggcacaac ctcaatggag tgatgcaacc
      421 tgcctggagt aaatgatgac acaaggcaat tgacccacgc atgtatctat ctcattttct
      481 tacaccttct attaccttct gctctctctg atttggaaaa agctgaaaaa aaaggttgaa
      541 accagttccc tgaaattatt cccctacttg actaataagt atataaagac ggtaggtatt
      601 gattgtaatt ctgtaaatct atttcttaaa cttcttaaat tctactttta tagttagtct
      661 tttttttagt tttaaaacac caagaactta gtttcgaata aacacacata aacagaatta
      721 tcacaagttt gtacaaaaaa gcaggctccg aattcgccct tggatccatg acggggctgt
      781 acgagctggt gtggcgggtg ctgcacgcgc tgctctgtct gcaccgcacg ctcacctcct
      841 ggctccgcgt tcggttcggc acctggaact ggatctggcg gcgctgctgc cgcgccgcct
      901 ctgccgcggt cctagcgccg ctcggcttca cgctccgcaa gcccccggca gtcggcagga
      961 accgccgtca ccaccggcac ccgcgcgggg ggtcgtgcct ggcagccgca caccaccgga
     1021 tgcgctggcg cgcggacggt cgttccttgg agaagctgcc tgtgcatatg ggcctggtga
     1081 tcaccgaggt ggagcaggaa cccagcttct cggacatcgc gagcctcgtg gtgtggtgta
     1141 tggccgtggg catctcctac attagcgtct acgaccacca aggtattttc aaaagaaata
     1201 attccagatt gatggatgaa attttaaaac aacagcaaga acttctgggc ctagattgtt
     1261 caaaatactc accagaattt gcaaatagta atgacaaaga tgatcaagtt ttaaattgcc
     1321 atttggcagt gaaggtgctg tctccggaag atggaaaagc agatattgta agagctgctc
     1381 aggacttttg ccagttagta gcccagaagc aaaagagacc cacagatttg gatgtagata
     1441 cgttagccag tttacttagt tcaaatggtt gtcctgatcc tgatttagta ttgaagttcg
     1501 gtcctgtgga cagcacatta ggctttcttc cctggcacat cagattgact gagattgtct
     1561 ctttgccttc ccacctaaac atcagttatg aggacttttt ctctgccctt cgtcaatatg
     1621 cagcctgtga acagcgtctg ggaaagtagg cggccgcaag ggcgaattcg acccagcttt
     1681 cttgtacaaa gtggtgattc gacttggttg aacacgttgc caaggcttaa gtgaatttac
     1741 tttaaatctt gcatttaaat aaattttctt tttatagctt tatgacttag tttcaattta
     1801 tatactattt taatgacatt ttcgattcgg atccgtgcat taatgaatcg gccaacgcgc
     1861 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1921 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1981 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2041 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2101 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2161 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2221 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2281 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2341 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2401 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2461 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2521 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2581 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2641 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2701 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2761 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2821 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2881 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2941 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3001 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3061 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3121 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3181 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3241 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3301 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3361 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3421 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3481 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3541 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3601 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3661 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3721 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3781 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     3841 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     3901 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3961 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     4021 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatactag
     4081 aaagtatagg aacttcagag cgcttttgaa aaccaaaagc gctctgaaga cgcactttca
     4141 aaaaaccaaa aacgcaccgg actgtaacga gctactaaaa tattgcgaat accgcttcca
     4201 caaacattgc tcaaaagtat ctctttgcta tatatctctg tgctatatcc ctatataacc
     4261 tacccatcca cctttcgctc cttgaacttg catctaaact cgacctctac attttttatg
     4321 tttatctcta gtattactct ttagacaaaa aaattgtagt aagaactatt catagagtga
     4381 atcgaaaaca atacgaaaat gtaaacattt cctatacgta gtatatagag acaaaataga
     4441 agaaaccgtt cataattttc tgaccaatga agaatcatca acgctatcac tttctgttca
     4501 caaagtatgc gcaatccaca tcggtataga atataatcgg ggatgccttt atcttgaaaa
     4561 aatgcacccg cagcttcgct agtaatcagt aaacgcggga agtggagtca ggcttttttt
     4621 atggaagaga aaatagacac caaagtagcc ttcttctaac cttaacggac ctacagtgca
     4681 aaaagttatc aagagactgc attatagagc gcacaaagga gaaaaaaagt aatctaagat
     4741 gctttgttag aaaaatagcg ctctcgggat gcatttttgt agaacaaaaa agaagtatag
     4801 attctttgtt ggtaaaatag cgctctcgcg ttgcatttct gttctgtaaa aatgcagctc
     4861 agattctttg tttgaaaaat tagcgctctc gcgttgcatt tttgttttac aaaaatgaag
     4921 cacagattct tcgttggtaa aatagcgctt tcgcgttgca tttctgttct gtaaaaatgc
     4981 agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt tctacaaaat
     5041 gaagcacaga tgcttcgtta actgtgggaa tactcaggta tcgtaagatg caagagttcg
     5101 aatctcttag caaccattat ttttttcctc aacataacga gaacacacag gggcgctatc
     5161 gcacagaatc aaattcgatg actggaaatt ttttgttaat ttcagaggtc gcctgacgca
     5221 tatacctttt tcaactgaaa aattgggaga aaaaggaaag gtgagagcgc cggaaccggc
     5281 ttttcatata gaatagagaa gcgttcatga ctaaatgctt gcatcacaat acttgaagtt
     5341 gacaatatta tttaaggacc tattgttttt tccaataggt ggttagcaat cgtcttactt
     5401 tctaactttt cttacctttt acatttcagc aatatatata tatatttcaa ggatatacca
     5461 ttctaatgtc tgcccctaag aagatcgtcg ttttgccagg tgaccataat atactagtaa
     5521 cgtaaatact agttagtaga tgatagttga tttttattcc aacactaaga aataatttcg
     5581 ccatttcttg aatgtattta aagatattta atgctataat agacatttaa atccaattct
     5641 tccaacatac aatgggagtt tggccgagtg gtttaaggcg tcagatttag gtggatttaa
     5701 cctctaaaat ctctgatatc ttcggatgca agggttcgaa tcccttagct ctcattattt
     5761 tttgcttttt ctcttgaggt cacatgatcg caaaatggca aatggcacgt gaagctgtcg
     5821 atattgggga actgtggtgg ttggcaaatg actaattaag ttagtcaagg cgccatcctc
     5881 atgaaaactg tgtaacataa taaccgaagt gtcgaaaagg tggcaccttg tccaattgaa
     5941 cacgctcgat gaaaaaataa gatatatata aggttaagta aagcgtctgt tagaaaggaa
     6001 gtttttcctt tttcttgctc tcttgtcttt tcatctacta tttccttcgt gtaatacagg
     6061 gtcgtcagat acatagatac aattctatta cccccatcca tacaatgcca tctcatttcg
     6121 atactgttca actacacgcc ggccaagaga accctggtga caatgctcac agatccagag
     6181 ctgtaccaat ttacgccacc acttcttatg ttttcgaaaa ctctaagcat ggttcgcaat
     6241 tgtttggtct agaagttcca ggttacgtct attcccgttt ccaaaaccca accagtaatg
     6301 ttttggaaga aagaattgct gctttagaag gtggtgctgc tgctttggct gtttcctccg
     6361 gtcaagccgc tcaaaccctt gccatccaag gtttggcaca cactggtgac aacatcgttt
     6421 ccacttctta cttatacggt ggtacttata accagttcaa aatctcgttc aaaagatttg
     6481 gtatcgaggc tagatttgtt gaaggtgaca atccagaaga attcgaaaag gtctttgatg
     6541 aaagaaccaa ggctgtttat ttggaaacca ttggtaatcc aaagtacaat gttccggatt
     6601 ttgaaaaaat tgttgcaatt gctcacaaac acggtattcc agttgtcgtt gacaacacat
     6661 ttggtgccgg tggttacttc tgtcagccaa ttaaatacgg tgctgatatt gtaacacatt
     6721 ctgctaccaa atggattggt ggtcatggta ctactatcgg tggtattatt gttgactctg
     6781 gtaagttccc atggaaggac tacccagaaa agttccctca attctctcaa cctgccgaag
     6841 gatatcacag tactatctac aatgaagcct acggtaactt ggcatacatc gttcatgtta
     6901 gaactgaact attaagagat ttgggtccat tgatgaaccc atttgcctct ttcttgctac
     6961 tacaaggtgt tgaaacatta tctttgagag ctgaaagaca cggtgaaaat gcattgaagt
     7021 tagccaaatg gttagaacaa tccccatacg tatcttgggt ttcataccct ggtttagcat
     7081 ctcattctca tcatgaaaat gctaggaagt atctatctaa cggtttcggt ggtgtcttat
     7141 ctttcggtgt aaaagactta ccaaatgccg acaaggaaac tgacccattc aaactttctg
     7201 gtgctcaagt tgttgacaat ttaaagcttg cctctaactt ggccaatgtt ggtgatgcca
     7261 agaccttagt cattgctcca tacttcacta cccacaaaca attaaatgac aaagaaaagt
     7321 tggcatctgg tgttaccaag gacttaattc gtgtctctgt tggtatcgaa tttattgatg
     7381 acattattgc agacttccag caatcttttg aaactgtttt cgctggccaa aaaccatgag
     7441 tgtgcgtaat gagttgtaaa attatgtata aacctacttt ctctcacaag tactatactt
     7501 ttataaaacg aactttattg aaatgaatat cctttttttc ccttgttaca tgtcgtgact
     7561 cgtactttga acctaaattg ttctaacatc aaagaacagt gttaattcgc agtcgagaag
     7621 aaaaatatgg tgaacaagac tcatctactt catgagacta ctttacgcct cctataaagc
     7681 tgtcacactg tacataaact ttataaatga aattcataat agaaacgaca cgaaattaca
     7741 aaatggaata tgttcatagg gtagacgaaa ctatatacgc aatctacata catttatcaa
     7801 gaaggagaaa aaggaggatg taaaggaata caggtaagca aattgatact aatggctcaa
     7861 cgtgataagg aaaaagaatt gcactttaac attaatattg acaaggagga gggcaccaca
     7921 caaaaagtta ggtgtaacag aaaatcatga aactatgatt cctaatttat atattggagg
     7981 attttctcta aaaaaaaaaa aatacaacaa ataaaaaaca ctcaatgacc tgaccatttg
     8041 atggagttta agtcaatacc ttcttgaacc atttcccata atggtgaaag ttccctcaag
     8101 aattttactc tgtcagaaac ggccttaacg acgtaccgga tctatgcggt gtgaaatacc
     8161 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     8221 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagc
//
