LOCUS       sequence-400040-        7143 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression vector for yeast RER2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7143)
  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 7143)
  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-GW1-RER2
FEATURES             Location/Qualifiers
     source          1..7143
                     /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(1650..1674)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1879..1896)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2050..2638)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2130..2149)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2809..3600,3601..3669))
                     /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     3432..3451
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3670..3774)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3842..3860
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3898..3920)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4020..4039
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4056..5038
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5039..5444
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5445..6539
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
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 tctcgagatg gaaacggata
      781 gtggtatacc tggtcattca tttgtgttaa agtggacaaa aaacatcttt tcgcgcacat
      841 tgcgtgcatc taactgtgta cctagacatg ttgggttcat catggatggg aacaggagat
      901 tcgctagaaa gaaagagatg gacgtaaagg agggccacga ggcaggattt gttagtatga
      961 gtagaatctt agaactgtgt tatgaagcag gagtcgatac ggctaccgtg tttgcctttt
     1021 caattgaaaa tttcaagagg agctcacggg aagttgaatc actgatgact ttagcgcgcg
     1081 aaaggatacg acaaatcaca gaacgtggag agctggcctg taagtatggg gtacgcatta
     1141 aaattatcgg cgatctctct ttgttggata agtctctatt agaagatgtt cgggttgctg
     1201 tggaaactac aaagaacaac aaaagggcca cgttaaatat ctgctttcca tatacaggca
     1261 gggaagaaat cttgcatgcc atgaaagaaa caattgttca acataagaag ggcgccgcta
     1321 tagacgaaag cacgttagaa tcgcatctct acacggcggg ggtaccccct ttagatttat
     1381 tgattaggac aagtggcgtt tccagattaa gtgacttttt gatatggcag gcatcgagta
     1441 agggcgtacg catcgaattg ctggattgtt tatggccaga gtttggacct atacggatgg
     1501 catggatttt attaaaattt tcgtttcaca aatccttttt aaacaaagag tacagattag
     1561 aggaaggtga ttatgacgag gaaaccaatg gggaccccat cgatttgaaa gaaaaaaagt
     1621 tgaattaagc ggccgcaagg gcgaattcga cccagctttc ttgtacaaag tggtgattcg
     1681 acttggttga acacgttgcc aaggcttaag tgaatttact ttaaatcttg catttaaata
     1741 aattttcttt ttatagcttt atgacttagt ttcaatttat atactatttt aatgacattt
     1801 tcgattcgga tccgtgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1861 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1921 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1981 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2041 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     2101 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     2161 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2221 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2281 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2341 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2401 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2461 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2521 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2581 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2641 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2701 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2761 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2821 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2881 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2941 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     3001 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     3061 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     3121 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     3181 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     3241 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3301 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3361 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3421 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3481 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3541 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3601 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3661 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3721 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3781 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     3841 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     3901 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     3961 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     4021 ggcatcagag cagattgtac tgagagtgca ccatactaga aagtatagga acttcagagc
     4081 gcttttgaaa accaaaagcg ctctgaagac gcactttcaa aaaaccaaaa acgcaccgga
     4141 ctgtaacgag ctactaaaat attgcgaata ccgcttccac aaacattgct caaaagtatc
     4201 tctttgctat atatctctgt gctatatccc tatataacct acccatccac ctttcgctcc
     4261 ttgaacttgc atctaaactc gacctctaca ttttttatgt ttatctctag tattactctt
     4321 tagacaaaaa aattgtagta agaactattc atagagtgaa tcgaaaacaa tacgaaaatg
     4381 taaacatttc ctatacgtag tatatagaga caaaatagaa gaaaccgttc ataattttct
     4441 gaccaatgaa gaatcatcaa cgctatcact ttctgttcac aaagtatgcg caatccacat
     4501 cggtatagaa tataatcggg gatgccttta tcttgaaaaa atgcacccgc agcttcgcta
     4561 gtaatcagta aacgcgggaa gtggagtcag gcttttttta tggaagagaa aatagacacc
     4621 aaagtagcct tcttctaacc ttaacggacc tacagtgcaa aaagttatca agagactgca
     4681 ttatagagcg cacaaaggag aaaaaaagta atctaagatg ctttgttaga aaaatagcgc
     4741 tctcgggatg catttttgta gaacaaaaaa gaagtataga ttctttgttg gtaaaatagc
     4801 gctctcgcgt tgcatttctg ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt
     4861 agcgctctcg cgttgcattt ttgttttaca aaaatgaagc acagattctt cgttggtaaa
     4921 atagcgcttt cgcgttgcat ttctgttctg taaaaatgca gctcagattc tttgtttgaa
     4981 aaattagcgc tctcgcgttg catttttgtt ctacaaaatg aagcacagat gcttcgttaa
     5041 ctgtgggaat actcaggtat cgtaagatgc aagagttcga atctcttagc aaccattatt
     5101 tttttcctca acataacgag aacacacagg ggcgctatcg cacagaatca aattcgatga
     5161 ctggaaattt tttgttaatt tcagaggtcg cctgacgcat ataccttttt caactgaaaa
     5221 attgggagaa aaaggaaagg tgagagcgcc ggaaccggct tttcatatag aatagagaag
     5281 cgttcatgac taaatgcttg catcacaata cttgaagttg acaatattat ttaaggacct
     5341 attgtttttt ccaataggtg gttagcaatc gtcttacttt ctaacttttc ttacctttta
     5401 catttcagca atatatatat atatttcaag gatataccat tctaatgtct gcccctaaga
     5461 agatcgtcgt tttgccaggt gaccacgttg gtcaagaaat cacagccgaa gccattaagg
     5521 ttcttaaagc tatttctgat gttcgttcca atgtcaagtt cgatttcgaa aatcatttaa
     5581 ttggtggtgc tgctatcgat gctacaggtg ttccacttcc agatgaggcg ctggaagcct
     5641 ccaagaaggc tgatgccgtt ttgttaggtg ctgtgggtgg tcctaaatgg ggtaccggta
     5701 gtgttagacc tgaacaaggt ttactaaaaa tccgtaaaga acttcaattg tacgccaact
     5761 taagaccatg taactttgca tccgactctc ttttagactt atctccaatc aagccacaat
     5821 ttgctaaagg tactgacttc gttgttgtca gagaattagt gggaggtatt tactttggta
     5881 agagaaagga agacgatggt gatggtgtcg cttgggatag tgaacaatac accgttccag
     5941 aagtgcaaag aatcacaaga atggccgctt tcatggccct acaacatgag ccaccattgc
     6001 ctatttggtc cttggataaa gctaatgttt tggcctcttc aagattatgg agaaaaactg
     6061 tggaggaaac catcaagaac gaattcccta cattgaaggt tcaacatcaa ttgattgatt
     6121 ctgccgccat gatcctagtt aagaacccaa cccacctaaa tggtattata atcaccagca
     6181 acatgtttgg tgatatcatc tccgatgaag cctccgttat cccaggttcc ttgggtttgt
     6241 tgccatctgc gtccttggcc tctttgccag acaagaacac cgcatttggt ttgtacgaac
     6301 catgccacgg ttctgctcca gatttgccaa agaataaggt caaccctatc gccactatct
     6361 tgtctgctgc aatgatgttg aaattgtcat tgaacttgcc tgaagaaggt aaggccattg
     6421 aagatgcagt taaaaaggtt ttggatgcag gtatcagaac tggtgattta ggtggttcca
     6481 acagtaccac cgaagtcggt gatgctgtcg ccgaagaagt taagaaaatc cttgcttaaa
     6541 aagattctct ttttttatga tatttgtaca taaactttat aaatgaaatt cataatagaa
     6601 acgacacgaa attacaaaat ggaatatgtt catagggtag acgaaactat atacgcaatc
     6661 tacatacatt tatcaagaag gagaaaaagg aggatgtaaa ggaatacagg taagcaaatt
     6721 gatactaatg gctcaacgtg ataaggaaaa agaattgcac tttaacatta atattgacaa
     6781 ggaggagggc accacacaaa aagttaggtg taacagaaaa tcatgaaact atgattccta
     6841 atttatatat tggaggattt tctctaaaaa aaaaaaaata caacaaataa aaaacactca
     6901 atgacctgac catttgatgg agtttaagtc aataccttct tgaaccattt cccataatgg
     6961 tgaaagttcc ctcaagaatt ttactctgtc agaaacggcc ttaacgacgt accggatcta
     7021 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     7081 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     7141 agc
//
