LOCUS       sequence-400043-        7179 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression vector for catalytic subunit of Methanosarcina 
            acetivorans cis-prenyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7179)
  AUTHORS   Grabinska KA, Edani BH, Park EJ, Kraehling JR, Sessa WC
  TITLE     A conserved C-terminal RXG motif in the NgBR subunit of 
            cis-prenyltransferase is critical for prenyltransferase activity.
  JOURNAL   J Biol Chem. 2017 Oct 20;292(42):17351-17361. doi: 
            10.1074/jbc.M117.806034. Epub 2017 Aug 23.
  PUBMED    28842490
REFERENCE   2  (bases 1 to 7179)
  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: 
            "10.1074/jbc.M117.806034"; journalName: "J Biol Chem"; date: 
            "2017-10-20- 20"; volume: "292"; issue: "42"; pages: "17351-17361"
COMMENT     Sequence Label: pKG-GW1-MaUPPS-A
FEATURES             Location/Qualifiers
     source          1..7179
                     /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(1686..1710)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1915..1932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2086..2674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2166..2185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2845..3636,3637..3705))
                     /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     3468..3487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3706..3810)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3878..3896
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3934..3956)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4056..4075
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      4092..5074
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        5075..5480
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             5481..6575
                     /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 taaacatatg aaaaacagaa
      781 cgtttagtat gttctatcag gaatatgagc agttactgga aaaagagata ctgtctagcg
      841 aaatacccga tcatatagct gtaataatgg acggcaatcg taggtacgct ggccaaaggg
      901 gcaggacaag gtccttcggc catgcgatgg gtgccgaagt gaccgaacag gtaattgaat
      961 ggtgctatga gattggggtg aaagagttga cactgtacgc ctttagcaca gaaaacttcc
     1021 agcgtagcga agaagaggta gacggtttat ttaacttaat caacgagaaa tttttaaagc
     1081 tgtataatga caagaggacg tatgagaagg agacccagat cagggtcatt ggtgacagaa
     1141 caaaattgcc cgcgttccta aacaagagta tcgaaaagat tgagaaagcc actgagacgc
     1201 atagaaaatt taatctgaac gtagccatag catacggagg taggcaggat ataatgcagg
     1261 cggtgcgtga catcgcggcc tgcgtgtcta gtgggaaatt gtctttagaa gacgttgacg
     1321 aaaacttaat atctaaacac ctatacccag cgccaggagt aagcgttcct aatgtagatt
     1381 tgatcgtaag gaccggcgga gatgaaagag tcagtaattt cctgccgtgg caagccaacg
     1441 gatctgagtg cgctacctac ttctgtgcgc cgttttggcc cgagtttaga aagatcgacc
     1501 tattgaggag tatcagggtg tatcaggcga ggaaggatga gaaaaagcag gagaattctt
     1561 atagggtctc aaaagttata aactttcttg gcgttggcaa atatggtgag aaaagcgagg
     1621 agttgggtca attactgccc ttaaagaagc aaggagttgc ctgagcggcc gcaagggcga
     1681 attcgaccca gctttcttgt acaaagtggt gattcgactt ggttgaacac gttgccaagg
     1741 cttaagtgaa tttactttaa atcttgcatt taaataaatt ttctttttat agctttatga
     1801 cttagtttca atttatatac tattttaatg acattttcga ttcggatccg tgcattaatg
     1861 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1921 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1981 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2041 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2101 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2161 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2221 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2281 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2341 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2401 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2461 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2521 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2581 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2641 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2701 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2761 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2821 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2881 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2941 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3001 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3061 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3121 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3181 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3241 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3301 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3361 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3421 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3481 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3541 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3601 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3661 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3721 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3781 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     3841 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     3901 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     3961 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     4021 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     4081 agtgcaccat actagaaagt ataggaactt cagagcgctt ttgaaaacca aaagcgctct
     4141 gaagacgcac tttcaaaaaa ccaaaaacgc accggactgt aacgagctac taaaatattg
     4201 cgaataccgc ttccacaaac attgctcaaa agtatctctt tgctatatat ctctgtgcta
     4261 tatccctata taacctaccc atccaccttt cgctccttga acttgcatct aaactcgacc
     4321 tctacatttt ttatgtttat ctctagtatt actctttaga caaaaaaatt gtagtaagaa
     4381 ctattcatag agtgaatcga aaacaatacg aaaatgtaaa catttcctat acgtagtata
     4441 tagagacaaa atagaagaaa ccgttcataa ttttctgacc aatgaagaat catcaacgct
     4501 atcactttct gttcacaaag tatgcgcaat ccacatcggt atagaatata atcggggatg
     4561 cctttatctt gaaaaaatgc acccgcagct tcgctagtaa tcagtaaacg cgggaagtgg
     4621 agtcaggctt tttttatgga agagaaaata gacaccaaag tagccttctt ctaaccttaa
     4681 cggacctaca gtgcaaaaag ttatcaagag actgcattat agagcgcaca aaggagaaaa
     4741 aaagtaatct aagatgcttt gttagaaaaa tagcgctctc gggatgcatt tttgtagaac
     4801 aaaaaagaag tatagattct ttgttggtaa aatagcgctc tcgcgttgca tttctgttct
     4861 gtaaaaatgc agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt
     4921 tttacaaaaa tgaagcacag attcttcgtt ggtaaaatag cgctttcgcg ttgcatttct
     4981 gttctgtaaa aatgcagctc agattctttg tttgaaaaat tagcgctctc gcgttgcatt
     5041 tttgttctac aaaatgaagc acagatgctt cgttaactgt gggaatactc aggtatcgta
     5101 agatgcaaga gttcgaatct cttagcaacc attatttttt tcctcaacat aacgagaaca
     5161 cacaggggcg ctatcgcaca gaatcaaatt cgatgactgg aaattttttg ttaatttcag
     5221 aggtcgcctg acgcatatac ctttttcaac tgaaaaattg ggagaaaaag gaaaggtgag
     5281 agcgccggaa ccggcttttc atatagaata gagaagcgtt catgactaaa tgcttgcatc
     5341 acaatacttg aagttgacaa tattatttaa ggacctattg ttttttccaa taggtggtta
     5401 gcaatcgtct tactttctaa cttttcttac cttttacatt tcagcaatat atatatatat
     5461 ttcaaggata taccattcta atgtctgccc ctaagaagat cgtcgttttg ccaggtgacc
     5521 acgttggtca agaaatcaca gccgaagcca ttaaggttct taaagctatt tctgatgttc
     5581 gttccaatgt caagttcgat ttcgaaaatc atttaattgg tggtgctgct atcgatgcta
     5641 caggtgttcc acttccagat gaggcgctgg aagcctccaa gaaggctgat gccgttttgt
     5701 taggtgctgt gggtggtcct aaatggggta ccggtagtgt tagacctgaa caaggtttac
     5761 taaaaatccg taaagaactt caattgtacg ccaacttaag accatgtaac tttgcatccg
     5821 actctctttt agacttatct ccaatcaagc cacaatttgc taaaggtact gacttcgttg
     5881 ttgtcagaga attagtggga ggtatttact ttggtaagag aaaggaagac gatggtgatg
     5941 gtgtcgcttg ggatagtgaa caatacaccg ttccagaagt gcaaagaatc acaagaatgg
     6001 ccgctttcat ggccctacaa catgagccac cattgcctat ttggtccttg gataaagcta
     6061 atgttttggc ctcttcaaga ttatggagaa aaactgtgga ggaaaccatc aagaacgaat
     6121 tccctacatt gaaggttcaa catcaattga ttgattctgc cgccatgatc ctagttaaga
     6181 acccaaccca cctaaatggt attataatca ccagcaacat gtttggtgat atcatctccg
     6241 atgaagcctc cgttatccca ggttccttgg gtttgttgcc atctgcgtcc ttggcctctt
     6301 tgccagacaa gaacaccgca tttggtttgt acgaaccatg ccacggttct gctccagatt
     6361 tgccaaagaa taaggtcaac cctatcgcca ctatcttgtc tgctgcaatg atgttgaaat
     6421 tgtcattgaa cttgcctgaa gaaggtaagg ccattgaaga tgcagttaaa aaggttttgg
     6481 atgcaggtat cagaactggt gatttaggtg gttccaacag taccaccgaa gtcggtgatg
     6541 ctgtcgccga agaagttaag aaaatccttg cttaaaaaga ttctcttttt ttatgatatt
     6601 tgtacataaa ctttataaat gaaattcata atagaaacga cacgaaatta caaaatggaa
     6661 tatgttcata gggtagacga aactatatac gcaatctaca tacatttatc aagaaggaga
     6721 aaaaggagga tgtaaaggaa tacaggtaag caaattgata ctaatggctc aacgtgataa
     6781 ggaaaaagaa ttgcacttta acattaatat tgacaaggag gagggcacca cacaaaaagt
     6841 taggtgtaac agaaaatcat gaaactatga ttcctaattt atatattgga ggattttctc
     6901 taaaaaaaaa aaaatacaac aaataaaaaa cactcaatga cctgaccatt tgatggagtt
     6961 taagtcaata ccttcttgaa ccatttccca taatggtgaa agttccctca agaattttac
     7021 tctgtcagaa acggccttaa cgacgtaccg gatctatgcg gtgtgaaata ccgcacagat
     7081 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     7141 aagggcgatc ggtgcgggcc tcttcgctat tacgccagc
//
