LOCUS       sequence-400044-        8055 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast expression vector for accessory subunit of Methanosarcina 
            acetivorans cis-prenyltransferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8055)
  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 8055)
  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-GW2-MaUPPS-B
FEATURES             Location/Qualifiers
     source          1..8055
                     /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(1459..1483)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1688..1705)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1859..2447)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1939..1958)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2618..3409,3410..3478))
                     /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     3241..3260
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3479..3583)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3651..3669
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3707..3729)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3829..3848
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      3865..4847
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     promoter        4848..5253
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     promoter        5543..5892
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /label=MET17 promoter
                     /note="expression is repressed by methionine"
     CDS             5893..7227
                     /codon_start=1
                     /gene="S. cerevisiae MET17"
                     /gene_synonym="MET15; MET25"
                     /product="O-acetylserine and O-acetylhomoserine 
                     sulfhydrylase, required for sulfur amino acid synthesis"
                     /label=MET17
                     /translation="MPSHFDTVQLHAGQENPGDNAHRSRAVPIYATTSYVFENSKHGSQ
                     LFGLEVPGYVYSRFQNPTSNVLEERIAALEGGAAALAVSSGQAAQTLAIQGLAHTGDNI
                     VSTSYLYGGTYNQFKISFKRFGIEARFVEGDNPEEFEKVFDERTKAVYLETIGNPKYNV
                     PDFEKIVAIAHKHGIPVVVDNTFGAGGYFCQPIKYGADIVTHSATKWIGGHGTTIGGII
                     VDSGKFPWKDYPEKFPQFSQPAEGYHSTIYNEAYGNLAYIVHVRTELLRDLGPLMNPFA
                     SFLLLQGVETLSLRAERHGENALKLAKWLEQSPYVSWVSYPGLASHSHHENARKYLSNG
                     FGGVLSFGVKDLPNADKETDPFKLSGAQVVDNLKLASNLANVGDAKTLVIAPYFTTHKQ
                     LNDKEKLASGVTKDLIRVSVGIEFIDDIIADFQQSFETVFAGQKP"
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 tcatatggac ctattaagct
      781 tcggccatcc gctttatgaa aggtttctag catggcagat aacccgtagc accactaaga
      841 cgcccgccca cgtagcaata attctaaagg agactgatct attggaaccc aagggtatgg
      901 agaaactttt atcagctttg ctaatcttca agagatataa cgtggaactt gtatcaattt
      961 acgtcgatat actaaaagcc gagcaacctt tgaaaaccga acttgcgagc acactaggaa
     1021 ctagcattga ggagatcttc acctgcctga gtagcgagac ggggtacaaa attcacggtc
     1081 tagagggtga ggtaaagagt tctcgtcaag gtaaagattt cttcgtgtat gtcagtcttg
     1141 gcttcggggg ccgtggtgag ataacccgtg cggtactatc aatcctagag gaggtgaaga
     1201 cgggcagtgt cagtcctgag gaggtggacg agaagatgtt ggagtcccac ctactggtaa
     1261 aacatgaacc ggatataatg attagatcag gtggacaaaa gcttagcgat ttcctggtct
     1321 ggcagagtgt atattcagaa ctatacttta cggatgtgaa ttggaaggat gtaaggagaa
     1381 tagacctact acgtgttatt cgtgatttcc aaaaaaggca aagaaggtat gggaagtgag
     1441 gtaccaaggg cgaattcgac ccagctttct tgtacaaagt ggtgattcga cttggttgaa
     1501 cacgttgcca aggcttaagt gaatttactt taaatcttgc atttaaataa attttctttt
     1561 tatagcttta tgacttagtt tcaatttata tactatttta atgacatttt cgattcggat
     1621 ccgtgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     1681 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     1741 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     1801 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     1861 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     1921 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     1981 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     2041 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     2101 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     2161 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     2221 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     2281 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     2341 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     2401 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     2461 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     2521 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     2581 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     2641 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     2701 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     2761 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     2821 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     2881 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     2941 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     3001 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     3061 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     3121 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     3181 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     3241 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     3301 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     3361 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     3421 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     3481 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     3541 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     3601 tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta
     3661 tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     3721 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     3781 agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc
     3841 agattgtact gagagtgcac catactagaa agtataggaa cttcagagcg cttttgaaaa
     3901 ccaaaagcgc tctgaagacg cactttcaaa aaaccaaaaa cgcaccggac tgtaacgagc
     3961 tactaaaata ttgcgaatac cgcttccaca aacattgctc aaaagtatct ctttgctata
     4021 tatctctgtg ctatatccct atataaccta cccatccacc tttcgctcct tgaacttgca
     4081 tctaaactcg acctctacat tttttatgtt tatctctagt attactcttt agacaaaaaa
     4141 attgtagtaa gaactattca tagagtgaat cgaaaacaat acgaaaatgt aaacatttcc
     4201 tatacgtagt atatagagac aaaatagaag aaaccgttca taattttctg accaatgaag
     4261 aatcatcaac gctatcactt tctgttcaca aagtatgcgc aatccacatc ggtatagaat
     4321 ataatcgggg atgcctttat cttgaaaaaa tgcacccgca gcttcgctag taatcagtaa
     4381 acgcgggaag tggagtcagg ctttttttat ggaagagaaa atagacacca aagtagcctt
     4441 cttctaacct taacggacct acagtgcaaa aagttatcaa gagactgcat tatagagcgc
     4501 acaaaggaga aaaaaagtaa tctaagatgc tttgttagaa aaatagcgct ctcgggatgc
     4561 atttttgtag aacaaaaaag aagtatagat tctttgttgg taaaatagcg ctctcgcgtt
     4621 gcatttctgt tctgtaaaaa tgcagctcag attctttgtt tgaaaaatta gcgctctcgc
     4681 gttgcatttt tgttttacaa aaatgaagca cagattcttc gttggtaaaa tagcgctttc
     4741 gcgttgcatt tctgttctgt aaaaatgcag ctcagattct ttgtttgaaa aattagcgct
     4801 ctcgcgttgc atttttgttc tacaaaatga agcacagatg cttcgttaac tgtgggaata
     4861 ctcaggtatc gtaagatgca agagttcgaa tctcttagca accattattt ttttcctcaa
     4921 cataacgaga acacacaggg gcgctatcgc acagaatcaa attcgatgac tggaaatttt
     4981 ttgttaattt cagaggtcgc ctgacgcata tacctttttc aactgaaaaa ttgggagaaa
     5041 aaggaaaggt gagagcgccg gaaccggctt ttcatataga atagagaagc gttcatgact
     5101 aaatgcttgc atcacaatac ttgaagttga caatattatt taaggaccta ttgttttttc
     5161 caataggtgg ttagcaatcg tcttactttc taacttttct taccttttac atttcagcaa
     5221 tatatatata tatttcaagg atataccatt ctaatgtctg cccctaagaa gatcgtcgtt
     5281 ttgccaggtg accataatat actagtaacg taaatactag ttagtagatg atagttgatt
     5341 tttattccaa cactaagaaa taatttcgcc atttcttgaa tgtatttaaa gatatttaat
     5401 gctataatag acatttaaat ccaattcttc caacatacaa tgggagtttg gccgagtggt
     5461 ttaaggcgtc agatttaggt ggatttaacc tctaaaatct ctgatatctt cggatgcaag
     5521 ggttcgaatc ccttagctct cattattttt tgctttttct cttgaggtca catgatcgca
     5581 aaatggcaaa tggcacgtga agctgtcgat attggggaac tgtggtggtt ggcaaatgac
     5641 taattaagtt agtcaaggcg ccatcctcat gaaaactgtg taacataata accgaagtgt
     5701 cgaaaaggtg gcaccttgtc caattgaaca cgctcgatga aaaaataaga tatatataag
     5761 gttaagtaaa gcgtctgtta gaaaggaagt ttttcctttt tcttgctctc ttgtcttttc
     5821 atctactatt tccttcgtgt aatacagggt cgtcagatac atagatacaa ttctattacc
     5881 cccatccata caatgccatc tcatttcgat actgttcaac tacacgccgg ccaagagaac
     5941 cctggtgaca atgctcacag atccagagct gtaccaattt acgccaccac ttcttatgtt
     6001 ttcgaaaact ctaagcatgg ttcgcaattg tttggtctag aagttccagg ttacgtctat
     6061 tcccgtttcc aaaacccaac cagtaatgtt ttggaagaaa gaattgctgc tttagaaggt
     6121 ggtgctgctg ctttggctgt ttcctccggt caagccgctc aaacccttgc catccaaggt
     6181 ttggcacaca ctggtgacaa catcgtttcc acttcttact tatacggtgg tacttataac
     6241 cagttcaaaa tctcgttcaa aagatttggt atcgaggcta gatttgttga aggtgacaat
     6301 ccagaagaat tcgaaaaggt ctttgatgaa agaaccaagg ctgtttattt ggaaaccatt
     6361 ggtaatccaa agtacaatgt tccggatttt gaaaaaattg ttgcaattgc tcacaaacac
     6421 ggtattccag ttgtcgttga caacacattt ggtgccggtg gttacttctg tcagccaatt
     6481 aaatacggtg ctgatattgt aacacattct gctaccaaat ggattggtgg tcatggtact
     6541 actatcggtg gtattattgt tgactctggt aagttcccat ggaaggacta cccagaaaag
     6601 ttccctcaat tctctcaacc tgccgaagga tatcacagta ctatctacaa tgaagcctac
     6661 ggtaacttgg catacatcgt tcatgttaga actgaactat taagagattt gggtccattg
     6721 atgaacccat ttgcctcttt cttgctacta caaggtgttg aaacattatc tttgagagct
     6781 gaaagacacg gtgaaaatgc attgaagtta gccaaatggt tagaacaatc cccatacgta
     6841 tcttgggttt cataccctgg tttagcatct cattctcatc atgaaaatgc taggaagtat
     6901 ctatctaacg gtttcggtgg tgtcttatct ttcggtgtaa aagacttacc aaatgccgac
     6961 aaggaaactg acccattcaa actttctggt gctcaagttg ttgacaattt aaagcttgcc
     7021 tctaacttgg ccaatgttgg tgatgccaag accttagtca ttgctccata cttcactacc
     7081 cacaaacaat taaatgacaa agaaaagttg gcatctggtg ttaccaagga cttaattcgt
     7141 gtctctgttg gtatcgaatt tattgatgac attattgcag acttccagca atcttttgaa
     7201 actgttttcg ctggccaaaa accatgagtg tgcgtaatga gttgtaaaat tatgtataaa
     7261 cctactttct ctcacaagta ctatactttt ataaaacgaa ctttattgaa atgaatatcc
     7321 tttttttccc ttgttacatg tcgtgactcg tactttgaac ctaaattgtt ctaacatcaa
     7381 agaacagtgt taattcgcag tcgagaagaa aaatatggtg aacaagactc atctacttca
     7441 tgagactact ttacgcctcc tataaagctg tcacactgta cataaacttt ataaatgaaa
     7501 ttcataatag aaacgacacg aaattacaaa atggaatatg ttcatagggt agacgaaact
     7561 atatacgcaa tctacataca tttatcaaga aggagaaaaa ggaggatgta aaggaataca
     7621 ggtaagcaaa ttgatactaa tggctcaacg tgataaggaa aaagaattgc actttaacat
     7681 taatattgac aaggaggagg gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa
     7741 ctatgattcc taatttatat attggaggat tttctctaaa aaaaaaaaaa tacaacaaat
     7801 aaaaaacact caatgacctg accatttgat ggagtttaag tcaatacctt cttgaaccat
     7861 ttcccataat ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg ccttaacgac
     7921 gtaccggatc tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     7981 ggcgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
     8041 cgctattacg ccagc
//
