LOCUS       sequence-400039-        8513 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression vector for yNUS1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8513)
  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 8513)
  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-NUS1
FEATURES             Location/Qualifiers
     source          1..8513
                     /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(1917..1941)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2146..2163)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2317..2905)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2397..2416)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3076..3867,3868..3936))
                     /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     3699..3718
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3937..4041)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4109..4127
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4165..4187)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4287..4306
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4323..5305
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5306..5711
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        6001..6350
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             6351..7685
                     /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 tgaattcatg cccacgatga
      781 tcaaaaagga tgataaagca atggagcccc ctaatgaaaa accgcataga aagatcgaaa
      841 gagatgatgt tccagaatct tccaatcaca tcccacctcc agaatctggt gttttaaagg
      901 gcggtaaagt taattcaaaa accagagctt taaaggccgt tacaagtatc attgcagacg
      961 ccgatgagaa ccctcagaag aaagtgaaca atgagacgaa tggagtccaa aagcaaaaga
     1021 cagaagattt gagtaaaaga ataggtaaat ttgaatacct tttttacaag tttttacttg
     1081 tgttgttata catctgcttc gggttgtttc ggtacggtca ataccaatat aataaaatga
     1141 aactaagaat attcagtatc atctacaacc atgcatatac accacagttg attagacagg
     1201 acgttattcc tctgaaaaaa attcctaaaa ggttggccgc tatcttggaa gtcaagccag
     1261 ttggcgacgt tggcggcggt gtgacaggtt tattaaatga cgcgagtgaa attgtttgct
     1321 ggactgtttc agctggtata aaacatttga tgttgtacga ttacgatgga atattacaaa
     1381 gaaatgttcc agagctgaga atggaaattc attccaacct ggctaaatat tttgggccag
     1441 ctcatgttcc aaactacgct gttaaaatac ctcattctaa caagatattc tacaatctag
     1501 acggaattga aaccgagact gatgtaggca atgagataga agctaaccaa gaaaaggaca
     1561 aaattgctat tgaaatttct ttattgtcta acagagatgg tagagaaacg attgtcgatc
     1621 tgaccaaaac tatggctgag ttatgtgcgg ttaacgaatt gagcgtttct gacatcacaa
     1681 tggatttagt tgattcagaa ctgaaacaac tagttggacc cgaaccagat ttactgttat
     1741 acttcgggcc ttcgttggat ttacaagggt tcccaccttg gcatattaga ttaaccgaat
     1801 tttattggga aaaagataac aacgaagtca tatattcggt tttcatccgc ggcctaagac
     1861 agtacgcagg atgtaaagtg aatgttggta aatgagcggc cgcaagggcg aattcgaccc
     1921 agctttcttg tacaaagtgg tgattcgact tggttgaaca cgttgccaag gcttaagtga
     1981 atttacttta aatcttgcat ttaaataaat tttcttttta tagctttatg acttagtttc
     2041 aatttatata ctattttaat gacattttcg attcggatcc gtgcattaat gaatcggcca
     2101 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     2161 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2221 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2281 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2341 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2401 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2461 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2521 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2581 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2641 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2701 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     2761 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2821 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2881 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2941 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     3001 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     3061 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     3121 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     3181 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     3241 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     3301 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     3361 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     3421 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     3481 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     3541 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     3601 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     3661 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     3721 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     3781 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     3841 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     3901 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     3961 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     4021 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     4081 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     4141 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     4201 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     4261 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     4321 tactagaaag tataggaact tcagagcgct tttgaaaacc aaaagcgctc tgaagacgca
     4381 ctttcaaaaa accaaaaacg caccggactg taacgagcta ctaaaatatt gcgaataccg
     4441 cttccacaaa cattgctcaa aagtatctct ttgctatata tctctgtgct atatccctat
     4501 ataacctacc catccacctt tcgctccttg aacttgcatc taaactcgac ctctacattt
     4561 tttatgttta tctctagtat tactctttag acaaaaaaat tgtagtaaga actattcata
     4621 gagtgaatcg aaaacaatac gaaaatgtaa acatttccta tacgtagtat atagagacaa
     4681 aatagaagaa accgttcata attttctgac caatgaagaa tcatcaacgc tatcactttc
     4741 tgttcacaaa gtatgcgcaa tccacatcgg tatagaatat aatcggggat gcctttatct
     4801 tgaaaaaatg cacccgcagc ttcgctagta atcagtaaac gcgggaagtg gagtcaggct
     4861 ttttttatgg aagagaaaat agacaccaaa gtagccttct tctaacctta acggacctac
     4921 agtgcaaaaa gttatcaaga gactgcatta tagagcgcac aaaggagaaa aaaagtaatc
     4981 taagatgctt tgttagaaaa atagcgctct cgggatgcat ttttgtagaa caaaaaagaa
     5041 gtatagattc tttgttggta aaatagcgct ctcgcgttgc atttctgttc tgtaaaaatg
     5101 cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg ttttacaaaa
     5161 atgaagcaca gattcttcgt tggtaaaata gcgctttcgc gttgcatttc tgttctgtaa
     5221 aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgttcta
     5281 caaaatgaag cacagatgct tcgttaactg tgggaatact caggtatcgt aagatgcaag
     5341 agttcgaatc tcttagcaac cattattttt ttcctcaaca taacgagaac acacaggggc
     5401 gctatcgcac agaatcaaat tcgatgactg gaaatttttt gttaatttca gaggtcgcct
     5461 gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa ggaaaggtga gagcgccgga
     5521 accggctttt catatagaat agagaagcgt tcatgactaa atgcttgcat cacaatactt
     5581 gaagttgaca atattattta aggacctatt gttttttcca ataggtggtt agcaatcgtc
     5641 ttactttcta acttttctta ccttttacat ttcagcaata tatatatata tttcaaggat
     5701 ataccattct aatgtctgcc cctaagaaga tcgtcgtttt gccaggtgac cataatatac
     5761 tagtaacgta aatactagtt agtagatgat agttgatttt tattccaaca ctaagaaata
     5821 atttcgccat ttcttgaatg tatttaaaga tatttaatgc tataatagac atttaaatcc
     5881 aattcttcca acatacaatg ggagtttggc cgagtggttt aaggcgtcag atttaggtgg
     5941 atttaacctc taaaatctct gatatcttcg gatgcaaggg ttcgaatccc ttagctctca
     6001 ttattttttg ctttttctct tgaggtcaca tgatcgcaaa atggcaaatg gcacgtgaag
     6061 ctgtcgatat tggggaactg tggtggttgg caaatgacta attaagttag tcaaggcgcc
     6121 atcctcatga aaactgtgta acataataac cgaagtgtcg aaaaggtggc accttgtcca
     6181 attgaacacg ctcgatgaaa aaataagata tatataaggt taagtaaagc gtctgttaga
     6241 aaggaagttt ttcctttttc ttgctctctt gtcttttcat ctactatttc cttcgtgtaa
     6301 tacagggtcg tcagatacat agatacaatt ctattacccc catccataca atgccatctc
     6361 atttcgatac tgttcaacta cacgccggcc aagagaaccc tggtgacaat gctcacagat
     6421 ccagagctgt accaatttac gccaccactt cttatgtttt cgaaaactct aagcatggtt
     6481 cgcaattgtt tggtctagaa gttccaggtt acgtctattc ccgtttccaa aacccaacca
     6541 gtaatgtttt ggaagaaaga attgctgctt tagaaggtgg tgctgctgct ttggctgttt
     6601 cctccggtca agccgctcaa acccttgcca tccaaggttt ggcacacact ggtgacaaca
     6661 tcgtttccac ttcttactta tacggtggta cttataacca gttcaaaatc tcgttcaaaa
     6721 gatttggtat cgaggctaga tttgttgaag gtgacaatcc agaagaattc gaaaaggtct
     6781 ttgatgaaag aaccaaggct gtttatttgg aaaccattgg taatccaaag tacaatgttc
     6841 cggattttga aaaaattgtt gcaattgctc acaaacacgg tattccagtt gtcgttgaca
     6901 acacatttgg tgccggtggt tacttctgtc agccaattaa atacggtgct gatattgtaa
     6961 cacattctgc taccaaatgg attggtggtc atggtactac tatcggtggt attattgttg
     7021 actctggtaa gttcccatgg aaggactacc cagaaaagtt ccctcaattc tctcaacctg
     7081 ccgaaggata tcacagtact atctacaatg aagcctacgg taacttggca tacatcgttc
     7141 atgttagaac tgaactatta agagatttgg gtccattgat gaacccattt gcctctttct
     7201 tgctactaca aggtgttgaa acattatctt tgagagctga aagacacggt gaaaatgcat
     7261 tgaagttagc caaatggtta gaacaatccc catacgtatc ttgggtttca taccctggtt
     7321 tagcatctca ttctcatcat gaaaatgcta ggaagtatct atctaacggt ttcggtggtg
     7381 tcttatcttt cggtgtaaaa gacttaccaa atgccgacaa ggaaactgac ccattcaaac
     7441 tttctggtgc tcaagttgtt gacaatttaa agcttgcctc taacttggcc aatgttggtg
     7501 atgccaagac cttagtcatt gctccatact tcactaccca caaacaatta aatgacaaag
     7561 aaaagttggc atctggtgtt accaaggact taattcgtgt ctctgttggt atcgaattta
     7621 ttgatgacat tattgcagac ttccagcaat cttttgaaac tgttttcgct ggccaaaaac
     7681 catgagtgtg cgtaatgagt tgtaaaatta tgtataaacc tactttctct cacaagtact
     7741 atacttttat aaaacgaact ttattgaaat gaatatcctt tttttccctt gttacatgtc
     7801 gtgactcgta ctttgaacct aaattgttct aacatcaaag aacagtgtta attcgcagtc
     7861 gagaagaaaa atatggtgaa caagactcat ctacttcatg agactacttt acgcctccta
     7921 taaagctgtc acactgtaca taaactttat aaatgaaatt cataatagaa acgacacgaa
     7981 attacaaaat ggaatatgtt catagggtag acgaaactat atacgcaatc tacatacatt
     8041 tatcaagaag gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt gatactaatg
     8101 gctcaacgtg ataaggaaaa agaattgcac tttaacatta atattgacaa ggaggagggc
     8161 accacacaaa aagttaggtg taacagaaaa tcatgaaact atgattccta atttatatat
     8221 tggaggattt tctctaaaaa aaaaaaaata caacaaataa aaaacactca atgacctgac
     8281 catttgatgg agtttaagtc aataccttct tgaaccattt cccataatgg tgaaagttcc
     8341 ctcaagaatt ttactctgtc agaaacggcc ttaacgacgt accggatcta tgcggtgtga
     8401 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     8461 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agc
//
