LOCUS       sequence-400042-        7080 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Yeast expression vector for Giardia cis-prenyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7080)
  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 7080)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-GlcisPT
FEATURES             Location/Qualifiers
     source          1..7080
                     /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(1587..1611)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1816..1833)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1987..2575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2067..2086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2746..3537,3538..3606))
                     /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     3369..3388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3607..3711)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3779..3797
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3835..3857)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3957..3976
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      3993..4975
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        4976..5381
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5382..6476
                     /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 tggatccatg atccccatgc
      781 atgtggcagt gataatggat ggcaatggtc gctgggctcg caagcagtta caggaacgga
      841 ccttcgggca cgaacagggc gtatctgttt tggagagcat cgttgatgaa tgtatcaact
      901 gcggaatccg ctttctcact gtttatgcgt tctctacaga aaattggagt cgtcccccta
      961 cagaggtttc atttcttttt gagctcctga gtgcggccat tcaacgcgta cggaccacct
     1021 acagagagag gaatgttaaa gttcaattct gtggagagcg aaccacacag ataccagaaa
     1081 ctgtcatcgc tgcaatgaac tgtattgaac aggacacagc tgcgtgtaca ggcctcatcc
     1141 tttccgtttg ctttaactat ggcggtcaca cagagatcgc acaagcctgt agaagcgtct
     1201 tagctgattg tctcgagggt gatgcagtgg agaacatcaa gacacggctt caaatgccaa
     1261 tagaacagtt catccaacag attgatacac acctctatgc aaatttgccc cctgtcgatc
     1321 ttcttatcag gacgggttgc gagaagcgac tctcgaattt cctcccctgg cacttagcct
     1381 atgctgaaat aatattcagc gaccttctct ggcctgaatt ctctgttcgg gccttcaaag
     1441 attgtctcga tgagtttgct tcgcgcacaa gacggttcgg aggggtccaa ctatccccaa
     1501 tgacgggcgt ttactcagac actcatcctc actcctccac aaatgcccta tcgaaccatg
     1561 actgagcggc cgcaagggcg aattcgaccc agctttcttg tacaaagtgg tgattcgact
     1621 tggttgaaca cgttgccaag gcttaagtga atttacttta aatcttgcat ttaaataaat
     1681 tttcttttta tagctttatg acttagtttc aatttatata ctattttaat gacattttcg
     1741 attcggatcc gtgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     1801 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1861 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1921 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1981 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     2041 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     2101 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     2161 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     2221 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     2281 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     2341 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     2401 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     2461 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2521 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2581 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2641 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2701 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2761 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2821 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2881 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2941 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     3001 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     3061 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     3121 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     3181 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     3241 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     3301 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     3361 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     3421 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     3481 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     3541 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3601 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3661 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3721 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3781 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     3841 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     3901 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     3961 atcagagcag attgtactga gagtgcacca tactagaaag tataggaact tcagagcgct
     4021 tttgaaaacc aaaagcgctc tgaagacgca ctttcaaaaa accaaaaacg caccggactg
     4081 taacgagcta ctaaaatatt gcgaataccg cttccacaaa cattgctcaa aagtatctct
     4141 ttgctatata tctctgtgct atatccctat ataacctacc catccacctt tcgctccttg
     4201 aacttgcatc taaactcgac ctctacattt tttatgttta tctctagtat tactctttag
     4261 acaaaaaaat tgtagtaaga actattcata gagtgaatcg aaaacaatac gaaaatgtaa
     4321 acatttccta tacgtagtat atagagacaa aatagaagaa accgttcata attttctgac
     4381 caatgaagaa tcatcaacgc tatcactttc tgttcacaaa gtatgcgcaa tccacatcgg
     4441 tatagaatat aatcggggat gcctttatct tgaaaaaatg cacccgcagc ttcgctagta
     4501 atcagtaaac gcgggaagtg gagtcaggct ttttttatgg aagagaaaat agacaccaaa
     4561 gtagccttct tctaacctta acggacctac agtgcaaaaa gttatcaaga gactgcatta
     4621 tagagcgcac aaaggagaaa aaaagtaatc taagatgctt tgttagaaaa atagcgctct
     4681 cgggatgcat ttttgtagaa caaaaaagaa gtatagattc tttgttggta aaatagcgct
     4741 ctcgcgttgc atttctgttc tgtaaaaatg cagctcagat tctttgtttg aaaaattagc
     4801 gctctcgcgt tgcatttttg ttttacaaaa atgaagcaca gattcttcgt tggtaaaata
     4861 gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct cagattcttt gtttgaaaaa
     4921 ttagcgctct cgcgttgcat ttttgttcta caaaatgaag cacagatgct tcgttaactg
     4981 tgggaatact caggtatcgt aagatgcaag agttcgaatc tcttagcaac cattattttt
     5041 ttcctcaaca taacgagaac acacaggggc gctatcgcac agaatcaaat tcgatgactg
     5101 gaaatttttt gttaatttca gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt
     5161 gggagaaaaa ggaaaggtga gagcgccgga accggctttt catatagaat agagaagcgt
     5221 tcatgactaa atgcttgcat cacaatactt gaagttgaca atattattta aggacctatt
     5281 gttttttcca ataggtggtt agcaatcgtc ttactttcta acttttctta ccttttacat
     5341 ttcagcaata tatatatata tttcaaggat ataccattct aatgtctgcc cctaagaaga
     5401 tcgtcgtttt gccaggtgac cacgttggtc aagaaatcac agccgaagcc attaaggttc
     5461 ttaaagctat ttctgatgtt cgttccaatg tcaagttcga tttcgaaaat catttaattg
     5521 gtggtgctgc tatcgatgct acaggtgttc cacttccaga tgaggcgctg gaagcctcca
     5581 agaaggctga tgccgttttg ttaggtgctg tgggtggtcc taaatggggt accggtagtg
     5641 ttagacctga acaaggttta ctaaaaatcc gtaaagaact tcaattgtac gccaacttaa
     5701 gaccatgtaa ctttgcatcc gactctcttt tagacttatc tccaatcaag ccacaatttg
     5761 ctaaaggtac tgacttcgtt gttgtcagag aattagtggg aggtatttac tttggtaaga
     5821 gaaaggaaga cgatggtgat ggtgtcgctt gggatagtga acaatacacc gttccagaag
     5881 tgcaaagaat cacaagaatg gccgctttca tggccctaca acatgagcca ccattgccta
     5941 tttggtcctt ggataaagct aatgttttgg cctcttcaag attatggaga aaaactgtgg
     6001 aggaaaccat caagaacgaa ttccctacat tgaaggttca acatcaattg attgattctg
     6061 ccgccatgat cctagttaag aacccaaccc acctaaatgg tattataatc accagcaaca
     6121 tgtttggtga tatcatctcc gatgaagcct ccgttatccc aggttccttg ggtttgttgc
     6181 catctgcgtc cttggcctct ttgccagaca agaacaccgc atttggtttg tacgaaccat
     6241 gccacggttc tgctccagat ttgccaaaga ataaggtcaa ccctatcgcc actatcttgt
     6301 ctgctgcaat gatgttgaaa ttgtcattga acttgcctga agaaggtaag gccattgaag
     6361 atgcagttaa aaaggttttg gatgcaggta tcagaactgg tgatttaggt ggttccaaca
     6421 gtaccaccga agtcggtgat gctgtcgccg aagaagttaa gaaaatcctt gcttaaaaag
     6481 attctctttt tttatgatat ttgtacataa actttataaa tgaaattcat aatagaaacg
     6541 acacgaaatt acaaaatgga atatgttcat agggtagacg aaactatata cgcaatctac
     6601 atacatttat caagaaggag aaaaaggagg atgtaaagga atacaggtaa gcaaattgat
     6661 actaatggct caacgtgata aggaaaaaga attgcacttt aacattaata ttgacaagga
     6721 ggagggcacc acacaaaaag ttaggtgtaa cagaaaatca tgaaactatg attcctaatt
     6781 tatatattgg aggattttct ctaaaaaaaa aaaaatacaa caaataaaaa acactcaatg
     6841 acctgaccat ttgatggagt ttaagtcaat accttcttga accatttccc ataatggtga
     6901 aagttccctc aagaatttta ctctgtcaga aacggcctta acgacgtacc ggatctatgc
     6961 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     7021 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
//
