LOCUS       sequence-430185-        7314 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression vector for yeast SRT1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7314)
  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 7314)
  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-SRT1
FEATURES             Location/Qualifiers
     source          1..7314
                     /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(1821..1845)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2050..2067)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2221..2809)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2301..2320)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2980..3771,3772..3840))
                     /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     3603..3622
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3841..3945)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4013..4031
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4069..4091)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4191..4210
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4227..5209
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5210..5615
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5616..6710
                     /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 tcccgggatg aaaatgccca
      781 gtattattca gattcagttt gtagccctaa aaaggctttt ggtagaaacc aaagaacaga
      841 tgtgcttcgc agtgaaaagt atatttcaga gagtatttgc gtgggttatg tcattaagct
      901 tgttttcatg gttttatgta aatcttcaga atattttgat aaaagcatta agggtagggc
      961 cagtgcctga acatgtctcc tttatcatgg atggtaaccg gagatatgcc aagtcaagaa
     1021 ggctaccagt aaaaaaaggc catgaagctg gtgggttaac gttactaaca ctactgtata
     1081 tctgcaaaag attgggtgta aaatgtgttt ccgcctatgc attttctatt gaaaatttta
     1141 atagaccaaa agaagaagta gatacgctaa tgaatttgtt tacggtaaag cttgatgaat
     1201 tcgcaaaaag agccaaggac tataaggatc ccttatacgg atctaaaata agaatagtag
     1261 gtgatcaatc tttactatct ccagaaatga gaaaaaaaat taaaaaagtg gaagaaatca
     1321 cacaggatgg agacgatttc actttattta tatgttttcc ttacacttca agaaatgata
     1381 tgttacatac tattcgtgat tcagttgaag accatttgga aaataaatca ccaaggatta
     1441 atataagaaa atttactaat aaaatgtaca tgggtttcca ttccaataaa tgtgaattat
     1501 taatcagaac aagtgggcat aggaggctct cagactatat gctatggcaa gtacatgaaa
     1561 atgccaccat tgaatttagt gatacgttgt ggccaaattt tagcttcttt gctatgtacc
     1621 tgatgattct caaatggtcc ttcttttcca ccattcaaaa atataatgag aagaatcact
     1681 cattgtttga aaaaatacat gaaagcgttc cttcaatatt taaaaaaaag aaaacagcta
     1741 tgtctttgta caactttcca aaccccccca tttcagtttc ggttacagga gatgaataag
     1801 cggccgcaag ggcgaattcg acccagcttt cttgtacaaa gtggtgattc gacttggttg
     1861 aacacgttgc caaggcttaa gtgaatttac tttaaatctt gcatttaaat aaattttctt
     1921 tttatagctt tatgacttag tttcaattta tatactattt taatgacatt ttcgattcgg
     1981 atccgtgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     2041 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     2101 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     2161 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2221 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2281 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2341 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2401 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2461 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2521 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2581 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2641 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2701 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2761 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2821 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     2881 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     2941 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     3001 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     3061 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     3121 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     3181 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     3241 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     3301 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     3361 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     3421 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     3481 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     3541 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     3601 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     3661 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     3721 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     3781 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     3841 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     3901 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     3961 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     4021 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4081 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4141 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     4201 gcagattgta ctgagagtgc accatactag aaagtatagg aacttcagag cgcttttgaa
     4261 aaccaaaagc gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg actgtaacga
     4321 gctactaaaa tattgcgaat accgcttcca caaacattgc tcaaaagtat ctctttgcta
     4381 tatatctctg tgctatatcc ctatataacc tacccatcca cctttcgctc cttgaacttg
     4441 catctaaact cgacctctac attttttatg tttatctcta gtattactct ttagacaaaa
     4501 aaattgtagt aagaactatt catagagtga atcgaaaaca atacgaaaat gtaaacattt
     4561 cctatacgta gtatatagag acaaaataga agaaaccgtt cataattttc tgaccaatga
     4621 agaatcatca acgctatcac tttctgttca caaagtatgc gcaatccaca tcggtataga
     4681 atataatcgg ggatgccttt atcttgaaaa aatgcacccg cagcttcgct agtaatcagt
     4741 aaacgcggga agtggagtca ggcttttttt atggaagaga aaatagacac caaagtagcc
     4801 ttcttctaac cttaacggac ctacagtgca aaaagttatc aagagactgc attatagagc
     4861 gcacaaagga gaaaaaaagt aatctaagat gctttgttag aaaaatagcg ctctcgggat
     4921 gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag cgctctcgcg
     4981 ttgcatttct gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc
     5041 gcgttgcatt tttgttttac aaaaatgaag cacagattct tcgttggtaa aatagcgctt
     5101 tcgcgttgca tttctgttct gtaaaaatgc agctcagatt ctttgtttga aaaattagcg
     5161 ctctcgcgtt gcatttttgt tctacaaaat gaagcacaga tgcttcgtta actgtgggaa
     5221 tactcaggta tcgtaagatg caagagttcg aatctcttag caaccattat ttttttcctc
     5281 aacataacga gaacacacag gggcgctatc gcacagaatc aaattcgatg actggaaatt
     5341 ttttgttaat ttcagaggtc gcctgacgca tatacctttt tcaactgaaa aattgggaga
     5401 aaaaggaaag gtgagagcgc cggaaccggc ttttcatata gaatagagaa gcgttcatga
     5461 ctaaatgctt gcatcacaat acttgaagtt gacaatatta tttaaggacc tattgttttt
     5521 tccaataggt ggttagcaat cgtcttactt tctaactttt cttacctttt acatttcagc
     5581 aatatatata tatatttcaa ggatatacca ttctaatgtc tgcccctaag aagatcgtcg
     5641 ttttgccagg tgaccacgtt ggtcaagaaa tcacagccga agccattaag gttcttaaag
     5701 ctatttctga tgttcgttcc aatgtcaagt tcgatttcga aaatcattta attggtggtg
     5761 ctgctatcga tgctacaggt gttccacttc cagatgaggc gctggaagcc tccaagaagg
     5821 ctgatgccgt tttgttaggt gctgtgggtg gtcctaaatg gggtaccggt agtgttagac
     5881 ctgaacaagg tttactaaaa atccgtaaag aacttcaatt gtacgccaac ttaagaccat
     5941 gtaactttgc atccgactct cttttagact tatctccaat caagccacaa tttgctaaag
     6001 gtactgactt cgttgttgtc agagaattag tgggaggtat ttactttggt aagagaaagg
     6061 aagacgatgg tgatggtgtc gcttgggata gtgaacaata caccgttcca gaagtgcaaa
     6121 gaatcacaag aatggccgct ttcatggccc tacaacatga gccaccattg cctatttggt
     6181 ccttggataa agctaatgtt ttggcctctt caagattatg gagaaaaact gtggaggaaa
     6241 ccatcaagaa cgaattccct acattgaagg ttcaacatca attgattgat tctgccgcca
     6301 tgatcctagt taagaaccca acccacctaa atggtattat aatcaccagc aacatgtttg
     6361 gtgatatcat ctccgatgaa gcctccgtta tcccaggttc cttgggtttg ttgccatctg
     6421 cgtccttggc ctctttgcca gacaagaaca ccgcatttgg tttgtacgaa ccatgccacg
     6481 gttctgctcc agatttgcca aagaataagg tcaaccctat cgccactatc ttgtctgctg
     6541 caatgatgtt gaaattgtca ttgaacttgc ctgaagaagg taaggccatt gaagatgcag
     6601 ttaaaaaggt tttggatgca ggtatcagaa ctggtgattt aggtggttcc aacagtacca
     6661 ccgaagtcgg tgatgctgtc gccgaagaag ttaagaaaat ccttgcttaa aaagattctc
     6721 tttttttatg atatttgtac ataaacttta taaatgaaat tcataataga aacgacacga
     6781 aattacaaaa tggaatatgt tcatagggta gacgaaacta tatacgcaat ctacatacat
     6841 ttatcaagaa ggagaaaaag gaggatgtaa aggaatacag gtaagcaaat tgatactaat
     6901 ggctcaacgt gataaggaaa aagaattgca ctttaacatt aatattgaca aggaggaggg
     6961 caccacacaa aaagttaggt gtaacagaaa atcatgaaac tatgattcct aatttatata
     7021 ttggaggatt ttctctaaaa aaaaaaaaat acaacaaata aaaaacactc aatgacctga
     7081 ccatttgatg gagtttaagt caataccttc ttgaaccatt tcccataatg gtgaaagttc
     7141 cctcaagaat tttactctgt cagaaacggc cttaacgacg taccggatct atgcggtgtg
     7201 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     7261 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagc
//
