LOCUS       sequence-469203-        3765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of GST-tagged PX domain (Cys at 2 and C85S) in 
            bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3765)
  AUTHORS   Buckles TC, Ohashi Y, Tremel S, McLaughlin SH, Pardon E, Steyaert J,
            Gordon MT, Williams RL, Falke JJ
  TITLE     The G-Protein Rab5A Activates VPS34 Complex II, a Class III PI3K, by
            a Dual Regulatory Mechanism.
  JOURNAL   Biophys J. 2020 Dec 1;119(11):2205-2218. doi: 
            10.1016/j.bpj.2020.10.028. Epub 2020 Oct 31.
  PUBMED    33137306
REFERENCE   2  (bases 1 to 3765)
  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.bpj.2020.10.028"; journalName: "Biophys J"; date: 
            "2020-12-1- 1"; volume: "119"; issue: "11"; pages: "2205-2218"
COMMENT     Sequence Label: pYO1645
FEATURES             Location/Qualifiers
     source          1..3765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(14..32)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        100..204
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(205..273,274..1065)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(423..442)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1236..1824
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1725..1744
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1978..1995
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2010..2149
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2075..2094)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2235..2257
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2310..2329
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2310..2328
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             2402..2424
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2435..3088
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     3046..3068
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             3107..3127
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(3673..3691)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3687..3734
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
       61 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
      121 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
      181 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
      241 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
      301 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
      361 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
      421 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
      481 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
      541 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
      601 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
      661 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
      721 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
      781 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
      841 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
      901 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
      961 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     1021 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     1081 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     1141 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     1201 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     1261 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     1321 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     1381 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     1441 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     1501 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     1561 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     1621 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     1681 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     1741 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     1801 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     1861 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     1921 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     1981 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     2041 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     2101 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     2161 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     2221 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     2281 cgcgaggcag gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg
     2341 gtttccctct agtgccggct ccggagagct ccaattggaa ttcgctagct ctagaaataa
     2401 ttttgtttaa ctttaagaag gagatataca taatatgtcc cctatactag gttattggaa
     2461 aattaagggc cttgtgcaac ccactcgact tcttttggaa tatcttgaag aaaaatatga
     2521 agagcatttg tatgagcgcg atgaaggtga taaatggcga aacaaaaagt ttgaattggg
     2581 tttggagttt cccaatcttc cttattatat tgatggtgat gttaaattaa cacagtctat
     2641 ggccatcata cgttatatag ctgacaagca caacatgttg ggtggttgtc caaaagagcg
     2701 tgcagagatt tcaatgcttg aaggagcggt tttggatatt agatacggtg tttcgagaat
     2761 tgcatatagt aaagactttg aaactctcaa agttgatttt cttagcaagc tacctgaaat
     2821 gctgaaaatg ttcgaagatc gtttatgtca taaaacatat ttaaatggtg atcatgtaac
     2881 ccatcctgac ttcatgttgt atgacgctct tgatgttgtt ttatacatgg acccaatgtg
     2941 cctggatgcg ttcccaaaat tagtttgttt taaaaaacgt attgaagcta tcccacaaat
     3001 tgataagtac ttgaaatcca gcaagtatat agcatggcct ttgcagggct ggcaagccac
     3061 gtttggtggt ggcgaccatc ctccaaaatc ggattcttct ggttccgaaa acctgtattt
     3121 ccagggctcc catatgtgcg ctgtggccca gcagctgcgg gccgagagtg actttgaaca
     3181 gcttccggat gatgttgcca tctcggccaa cattgctgac atcgaggaga agagaggctt
     3241 caccagccac tttgttttcg tcatcgaggt gaagacaaaa ggaggatcca agtacctcat
     3301 ctaccgccgc taccgccagt tccatgcttt gcagagcaag ctggaggagc gcttcgggcc
     3361 agacagcaag agcagtgccc tggccagtac cctgcccaca ctcccagcca aagtctacgt
     3421 gggtgtgaaa caggagatcg ccgagatgcg gatacctgcc ctcaacgcct acatgaagag
     3481 cctgctcagc ctgccggtct gggtgctgat ggatgaggac gtccggatct tcttttacca
     3541 gtcgccctat gactcagagc aggtgcccca ggccatccgc cggtgaggat ccagatctaa
     3601 gcttggtacc acgcgtgcgc gctgatccgg ctgctaacaa agcccgaaag gaagctgagt
     3661 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     3721 tgaggggttt tttgctgaaa ggaggaacta tagccggata attct
//
