LOCUS       sequence-400038-        7281 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression vector for human DHDDS.
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
FEATURES             Location/Qualifiers
     source          1..7281
                     /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(1788..1812)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2017..2034)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2188..2776)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2268..2287)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2947..3738,3739..3807))
                     /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     3570..3589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3808..3912)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3980..3998
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4036..4058)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4158..4177
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4194..5176
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5177..5582
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5583..6677
                     /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 tcatatgtca tggatcaagg
      781 aaggagagct gtcactttgg gagcggttct gtgccaacat cataaaggca ggcccaatgc
      841 cgaaacacat tgcattcata atggacggga accgtcgcta tgccaagaag tgccaggtgg
      901 agcggcagga aggccactca cagggcttca acaagctagc tgagactctg cggtggtgtt
      961 tgaacctggg catcctagag gtgacagtct acgcattcag cattgagaac ttcaaacgct
     1021 ccaagagtga ggtagacggg cttatggatc tggcccggca gaagttcagc cgcttgatgg
     1081 aagaaaagga gaaactgcag aagcatgggg tgtgtatccg ggtcctgggc gatctgcact
     1141 tgttgccctt ggatctccag gagctgattg cacaagctgt acaggccacg aagaactaca
     1201 acaagtgttt cctgaatgtc tgttttgcat acacatcccg tcatgagatc agcaatgctg
     1261 tgagagagat ggcctggggg gtggagcaag gcctgttgga tcccagtgat atctctgagt
     1321 ctctgcttga taagtgcctc tataccaacc gctctcctca tcctgacatc ttgatacgga
     1381 cttctggaga agtgcggctg agtgacttct tgctatggca gacctctcac tcctgcctgg
     1441 tgttccaacc cgttctgtgg ccagagtata cattttggaa cctcttcgag gccatcctgc
     1501 agttccagat gaaccatagc atgcttcaga aggcccgaga catgtatgca gaggagcgga
     1561 agaggcagca gctggagagg gaccaggcta cagtgacaga gcagctgctg cgagaggggc
     1621 tccaagccag tggggacgcc cagctccgaa ggacacgctt gcacaaactc tcggccagac
     1681 gggaagagcg agtccaaggc ttcctgcagg ccttggaact caagcgagct gactggctgg
     1741 cccgtctggg cactgcatca gcctgagcgg ccgcaagggc gaattcgacc cagctttctt
     1801 gtacaaagtg gtgattcgac ttggttgaac acgttgccaa ggcttaagtg aatttacttt
     1861 aaatcttgca tttaaataaa ttttcttttt atagctttat gacttagttt caatttatat
     1921 actattttaa tgacattttc gattcggatc cgtgcattaa tgaatcggcc aacgcgcggg
     1981 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2041 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2101 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2161 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2221 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2281 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2341 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2401 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2461 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2521 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2581 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     2641 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2701 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2761 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2821 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     2881 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     2941 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3001 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3061 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3121 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3181 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3241 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3301 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3361 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3421 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3481 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3541 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3601 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3661 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3721 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3781 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3841 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     3901 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     3961 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     4021 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     4081 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     4141 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atactagaaa
     4201 gtataggaac ttcagagcgc ttttgaaaac caaaagcgct ctgaagacgc actttcaaaa
     4261 aaccaaaaac gcaccggact gtaacgagct actaaaatat tgcgaatacc gcttccacaa
     4321 acattgctca aaagtatctc tttgctatat atctctgtgc tatatcccta tataacctac
     4381 ccatccacct ttcgctcctt gaacttgcat ctaaactcga cctctacatt ttttatgttt
     4441 atctctagta ttactcttta gacaaaaaaa ttgtagtaag aactattcat agagtgaatc
     4501 gaaaacaata cgaaaatgta aacatttcct atacgtagta tatagagaca aaatagaaga
     4561 aaccgttcat aattttctga ccaatgaaga atcatcaacg ctatcacttt ctgttcacaa
     4621 agtatgcgca atccacatcg gtatagaata taatcgggga tgcctttatc ttgaaaaaat
     4681 gcacccgcag cttcgctagt aatcagtaaa cgcgggaagt ggagtcaggc tttttttatg
     4741 gaagagaaaa tagacaccaa agtagccttc ttctaacctt aacggaccta cagtgcaaaa
     4801 agttatcaag agactgcatt atagagcgca caaaggagaa aaaaagtaat ctaagatgct
     4861 ttgttagaaa aatagcgctc tcgggatgca tttttgtaga acaaaaaaga agtatagatt
     4921 ctttgttggt aaaatagcgc tctcgcgttg catttctgtt ctgtaaaaat gcagctcaga
     4981 ttctttgttt gaaaaattag cgctctcgcg ttgcattttt gttttacaaa aatgaagcac
     5041 agattcttcg ttggtaaaat agcgctttcg cgttgcattt ctgttctgta aaaatgcagc
     5101 tcagattctt tgtttgaaaa attagcgctc tcgcgttgca tttttgttct acaaaatgaa
     5161 gcacagatgc ttcgttaact gtgggaatac tcaggtatcg taagatgcaa gagttcgaat
     5221 ctcttagcaa ccattatttt tttcctcaac ataacgagaa cacacagggg cgctatcgca
     5281 cagaatcaaa ttcgatgact ggaaattttt tgttaatttc agaggtcgcc tgacgcatat
     5341 acctttttca actgaaaaat tgggagaaaa aggaaaggtg agagcgccgg aaccggcttt
     5401 tcatatagaa tagagaagcg ttcatgacta aatgcttgca tcacaatact tgaagttgac
     5461 aatattattt aaggacctat tgttttttcc aataggtggt tagcaatcgt cttactttct
     5521 aacttttctt accttttaca tttcagcaat atatatatat atttcaagga tataccattc
     5581 taatgtctgc ccctaagaag atcgtcgttt tgccaggtga ccacgttggt caagaaatca
     5641 cagccgaagc cattaaggtt cttaaagcta tttctgatgt tcgttccaat gtcaagttcg
     5701 atttcgaaaa tcatttaatt ggtggtgctg ctatcgatgc tacaggtgtt ccacttccag
     5761 atgaggcgct ggaagcctcc aagaaggctg atgccgtttt gttaggtgct gtgggtggtc
     5821 ctaaatgggg taccggtagt gttagacctg aacaaggttt actaaaaatc cgtaaagaac
     5881 ttcaattgta cgccaactta agaccatgta actttgcatc cgactctctt ttagacttat
     5941 ctccaatcaa gccacaattt gctaaaggta ctgacttcgt tgttgtcaga gaattagtgg
     6001 gaggtattta ctttggtaag agaaaggaag acgatggtga tggtgtcgct tgggatagtg
     6061 aacaatacac cgttccagaa gtgcaaagaa tcacaagaat ggccgctttc atggccctac
     6121 aacatgagcc accattgcct atttggtcct tggataaagc taatgttttg gcctcttcaa
     6181 gattatggag aaaaactgtg gaggaaacca tcaagaacga attccctaca ttgaaggttc
     6241 aacatcaatt gattgattct gccgccatga tcctagttaa gaacccaacc cacctaaatg
     6301 gtattataat caccagcaac atgtttggtg atatcatctc cgatgaagcc tccgttatcc
     6361 caggttcctt gggtttgttg ccatctgcgt ccttggcctc tttgccagac aagaacaccg
     6421 catttggttt gtacgaacca tgccacggtt ctgctccaga tttgccaaag aataaggtca
     6481 accctatcgc cactatcttg tctgctgcaa tgatgttgaa attgtcattg aacttgcctg
     6541 aagaaggtaa ggccattgaa gatgcagtta aaaaggtttt ggatgcaggt atcagaactg
     6601 gtgatttagg tggttccaac agtaccaccg aagtcggtga tgctgtcgcc gaagaagtta
     6661 agaaaatcct tgcttaaaaa gattctcttt ttttatgata tttgtacata aactttataa
     6721 atgaaattca taatagaaac gacacgaaat tacaaaatgg aatatgttca tagggtagac
     6781 gaaactatat acgcaatcta catacattta tcaagaagga gaaaaaggag gatgtaaagg
     6841 aatacaggta agcaaattga tactaatggc tcaacgtgat aaggaaaaag aattgcactt
     6901 taacattaat attgacaagg aggagggcac cacacaaaaa gttaggtgta acagaaaatc
     6961 atgaaactat gattcctaat ttatatattg gaggattttc tctaaaaaaa aaaaaataca
     7021 acaaataaaa aacactcaat gacctgacca tttgatggag tttaagtcaa taccttcttg
     7081 aaccatttcc cataatggtg aaagttccct caagaatttt actctgtcag aaacggcctt
     7141 aacgacgtac cggatctatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     7201 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     7261 cctcttcgct attacgccag c
//
