LOCUS       sequence-405867-        7281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression vector for human DHDDS K42E.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7281)
  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 7281)
  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-hCIT K42E
FEATURES             Location/Qualifiers
     source          1..7281
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        59..725
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     595..618
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     protein_bind    728..752
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1793..1817)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2022..2039)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2193..2781)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2273..2292)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2952..3743,3744..3812))
                     /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     3575..3594
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3813..3917)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3985..4003
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4041..4063)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4163..4182
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4199..5181
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5182..5587
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5588..6682
                     /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 ccagcggatc cggtagaatc attttgaata aaaaacacgc tttttcagtt cgagtttatc
       61 attatcaata ctgccatttc aaagaatacg taaataatta atagtagtga ttttcctaac
      121 tttatttagt caaaaaatta gccttttaat tctgctgtaa cccgtacatg cccaaaatag
      181 ggggcgggtt acacagaata tataacatcg taggtgtctg ggtgaacagt ttattcctgg
      241 catccactaa atataatgga gcccgctttt taagctggca tccagaaaaa aaaagaatcc
      301 cagcaccaaa atattgtttt cttcaccaac catcagttca taggtccatt ctcttagcgc
      361 aactacagag aacaggggca caaacaggca aaaaacgggc acaacctcaa tggagtgatg
      421 caacctgcct ggagtaaatg atgacacaag gcaattgacc cacgcatgta tctatctcat
      481 tttcttacac cttctattac cttctgctct ctctgatttg gaaaaagctg aaaaaaaagg
      541 ttgaaaccag ttccctgaaa ttattcccct acttgactaa taagtatata aagacggtag
      601 gtattgattg taattctgta aatctatttc ttaaacttct taaattctac ttttatagtt
      661 agtctttttt ttagttttaa aacaccaaga acttagtttc gaataaacac acataaacag
      721 aattatcaca agtttgtaca aaaaagcagg ctccgaattc gcccttcata tgtcatggat
      781 caaggaagga gagctgtcac tttgggagcg gttctgtgcc aacatcataa aggcaggccc
      841 aatgccgaaa cacattgcat tcataatgga cgggaaccgt cgctatgcca aggagtgcca
      901 ggtggagcgg caggaaggcc actcacaggg cttcaacaag ctagctgaga ctctgcggtg
      961 gtgtttgaac ctgggcatcc tagaggtgac agtctacgca ttcagcattg agaacttcaa
     1021 acgctccaag agtgaggtag acgggcttat ggatctggcc cggcagaagt tcagccgctt
     1081 gatggaagaa aaggagaaac tgcagaagca tggggtgtgt atccgggtcc tgggcgatct
     1141 gcacttgttg cccttggatc tccaggagct gattgcacaa gctgtacagg ccacgaagaa
     1201 ctacaacaag tgtttcctga atgtctgttt tgcatacaca tcccgtcatg agatcagcaa
     1261 tgctgtgaga gagatggcct ggggggtgga gcaaggcctg ttggatccca gtgatatctc
     1321 tgagtctctg cttgataagt gcctctatac caaccgctct cctcatcctg acatcttgat
     1381 acggacttct ggagaagtgc ggctgagtga cttcttgcta tggcagacct ctcactcctg
     1441 cctggtgttc caacccgttc tgtggccaga gtatacattt tggaacctct tcgaggccat
     1501 cctgcagttc cagatgaacc atagcgtgct tcagaaggcc cgagacatgt atgcagagga
     1561 gcggaagagg cagcagctgg agagggacca ggctacagtg acagagcagc tgctgcgaga
     1621 ggggctccaa gccagtgggg acgcccagct ccgaaggaca cgcttgcaca aactctcggc
     1681 cagacgggaa gagcgagtcc aaggcttcct gcaggccttg gaactcaagc gagctgactg
     1741 gctggcccgt ctgggcactg catcagcctg agcggccgca agggcgaatt cgacccagct
     1801 ttcttgtaca aagtggtgat tcgacttggt tgaacacgtt gccaaggctt aagtgaattt
     1861 actttaaatc ttgcatttaa ataaattttc tttttatagc tttatgactt agtttcaatt
     1921 tatatactat tttaatgaca ttttcgattc ggatccgtgc attaatgaat cggccaacgc
     1981 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     2041 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     2101 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     2161 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     2221 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     2281 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     2341 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     2401 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     2461 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     2521 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     2581 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     2641 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2701 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2761 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2821 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2881 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2941 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     3001 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     3061 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     3121 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     3181 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3241 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3301 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3361 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3421 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3481 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3541 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3601 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3661 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3721 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3781 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3841 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3901 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     3961 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     4021 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     4081 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     4141 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatact
     4201 agaaagtata ggaacttcag agcgcttttg aaaaccaaaa gcgctctgaa gacgcacttt
     4261 caaaaaacca aaaacgcacc ggactgtaac gagctactaa aatattgcga ataccgcttc
     4321 cacaaacatt gctcaaaagt atctctttgc tatatatctc tgtgctatat ccctatataa
     4381 cctacccatc cacctttcgc tccttgaact tgcatctaaa ctcgacctct acatttttta
     4441 tgtttatctc tagtattact ctttagacaa aaaaattgta gtaagaacta ttcatagagt
     4501 gaatcgaaaa caatacgaaa atgtaaacat ttcctatacg tagtatatag agacaaaata
     4561 gaagaaaccg ttcataattt tctgaccaat gaagaatcat caacgctatc actttctgtt
     4621 cacaaagtat gcgcaatcca catcggtata gaatataatc ggggatgcct ttatcttgaa
     4681 aaaatgcacc cgcagcttcg ctagtaatca gtaaacgcgg gaagtggagt caggcttttt
     4741 ttatggaaga gaaaatagac accaaagtag ccttcttcta accttaacgg acctacagtg
     4801 caaaaagtta tcaagagact gcattataga gcgcacaaag gagaaaaaaa gtaatctaag
     4861 atgctttgtt agaaaaatag cgctctcggg atgcattttt gtagaacaaa aaagaagtat
     4921 agattctttg ttggtaaaat agcgctctcg cgttgcattt ctgttctgta aaaatgcagc
     4981 tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgtttt acaaaaatga
     5041 agcacagatt cttcgttggt aaaatagcgc tttcgcgttg catttctgtt ctgtaaaaat
     5101 gcagctcaga ttctttgttt gaaaaattag cgctctcgcg ttgcattttt gttctacaaa
     5161 atgaagcaca gatgcttcgt taactgtggg aatactcagg tatcgtaaga tgcaagagtt
     5221 cgaatctctt agcaaccatt atttttttcc tcaacataac gagaacacac aggggcgcta
     5281 tcgcacagaa tcaaattcga tgactggaaa ttttttgtta atttcagagg tcgcctgacg
     5341 catatacctt tttcaactga aaaattggga gaaaaaggaa aggtgagagc gccggaaccg
     5401 gcttttcata tagaatagag aagcgttcat gactaaatgc ttgcatcaca atacttgaag
     5461 ttgacaatat tatttaagga cctattgttt tttccaatag gtggttagca atcgtcttac
     5521 tttctaactt ttcttacctt ttacatttca gcaatatata tatatatttc aaggatatac
     5581 cattctaatg tctgccccta agaagatcgt cgttttgcca ggtgaccacg ttggtcaaga
     5641 aatcacagcc gaagccatta aggttcttaa agctatttct gatgttcgtt ccaatgtcaa
     5701 gttcgatttc gaaaatcatt taattggtgg tgctgctatc gatgctacag gtgttccact
     5761 tccagatgag gcgctggaag cctccaagaa ggctgatgcc gttttgttag gtgctgtggg
     5821 tggtcctaaa tggggtaccg gtagtgttag acctgaacaa ggtttactaa aaatccgtaa
     5881 agaacttcaa ttgtacgcca acttaagacc atgtaacttt gcatccgact ctcttttaga
     5941 cttatctcca atcaagccac aatttgctaa aggtactgac ttcgttgttg tcagagaatt
     6001 agtgggaggt atttactttg gtaagagaaa ggaagacgat ggtgatggtg tcgcttggga
     6061 tagtgaacaa tacaccgttc cagaagtgca aagaatcaca agaatggccg ctttcatggc
     6121 cctacaacat gagccaccat tgcctatttg gtccttggat aaagctaatg ttttggcctc
     6181 ttcaagatta tggagaaaaa ctgtggagga aaccatcaag aacgaattcc ctacattgaa
     6241 ggttcaacat caattgattg attctgccgc catgatccta gttaagaacc caacccacct
     6301 aaatggtatt ataatcacca gcaacatgtt tggtgatatc atctccgatg aagcctccgt
     6361 tatcccaggt tccttgggtt tgttgccatc tgcgtccttg gcctctttgc cagacaagaa
     6421 caccgcattt ggtttgtacg aaccatgcca cggttctgct ccagatttgc caaagaataa
     6481 ggtcaaccct atcgccacta tcttgtctgc tgcaatgatg ttgaaattgt cattgaactt
     6541 gcctgaagaa ggtaaggcca ttgaagatgc agttaaaaag gttttggatg caggtatcag
     6601 aactggtgat ttaggtggtt ccaacagtac caccgaagtc ggtgatgctg tcgccgaaga
     6661 agttaagaaa atccttgctt aaaaagattc tcttttttta tgatatttgt acataaactt
     6721 tataaatgaa attcataata gaaacgacac gaaattacaa aatggaatat gttcataggg
     6781 tagacgaaac tatatacgca atctacatac atttatcaag aaggagaaaa aggaggatgt
     6841 aaaggaatac aggtaagcaa attgatacta atggctcaac gtgataagga aaaagaattg
     6901 cactttaaca ttaatattga caaggaggag ggcaccacac aaaaagttag gtgtaacaga
     6961 aaatcatgaa actatgattc ctaatttata tattggagga ttttctctaa aaaaaaaaaa
     7021 atacaacaaa taaaaaacac tcaatgacct gaccatttga tggagtttaa gtcaatacct
     7081 tcttgaacca tttcccataa tggtgaaagt tccctcaaga attttactct gtcagaaacg
     7141 gccttaacga cgtaccggat ctatgcggtg tgaaataccg cacagatgcg taaggagaaa
     7201 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     7261 gcgggcctct tcgctattac g
//
