LOCUS       sequence-468451-        5337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of FL His6- human VPS34 in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5337)
  AUTHORS   Ohashi Y, Tremel S, Masson GR, McGinney L, Boulanger J, 
            Rostislavleva K, Johnson CM, Niewczas I, Clark J, Williams RL
  TITLE     Membrane characteristics tune activities of endosomal and autophagic
            human VPS34 complexes.
  JOURNAL   Elife. 2020 Jun 30;9:e58281. doi: 10.7554/eLife.58281.
  PUBMED    32602837
REFERENCE   2  (bases 1 to 5337)
  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"; journalName: "Elife."; 
            date: "2020-06-30"; volume: "9:e58281. doi"; pages: " 
            10.7554/eLife.58281"
COMMENT     Sequence Label: pSM41
FEATURES             Location/Qualifiers
     source          1..5337
                     /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             2438..2455
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2468..2488
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(5245..5263)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5259..5306
                     /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 gagatataaa tatggctcac catcatcatc atcattcttc
     2461 tggttccgaa aacctgtatt tccagggctc ccatatgggc gaagcagaga agtttcacta
     2521 catctatagt tgtgacctgg atatcaacgt ccagcttaag ataggaagct tggaagggaa
     2581 gagagaacaa aagagttata aagctgtcct ggaagaccca atgttgaagt tctcaggact
     2641 atatcaagag acatgctctg atctttatgt tacttgtcaa gtttttgcag aagggaagcc
     2701 tttggccttg ccagtgagaa catcctacaa agcatttagt acaagatgga actggaatga
     2761 atggctgaaa ctaccagtaa aataccctga cctgcccagg aatgcccaag tggccctcac
     2821 catctgggat gtgtatggtc ccggaaaagc agtgcctgta ggaggaacaa cggtttcgct
     2881 ctttggaaaa tacggcatgt ttcgccaagg gatgcatgac ttgaaagtct ggcctaatgt
     2941 agaagcagat ggatcagaac ccacaaaaac tcctggcaga acaagtagca ctctctcaga
     3001 agatcagatg agccgtcttg ccaagctcac caaagctcat cgacaaggac acatggtgaa
     3061 agtagattgg ctggatagat tgacatttag agaaatagaa atgataaatg agagtgaaaa
     3121 acgaagttct aatttcatgt acctgatggt tgaatttcga tgtgtcaagt gtgatgataa
     3181 ggaatatggt attgtttatt atgaaaagga cggtgatgaa tcatctccaa ttttaacaag
     3241 ttttgaatta gtgaaagttc ctgaccccca gatgtctatg gagaatttag ttgagagcaa
     3301 acaccacaag cttgcccgga gtttaagaag tggaccttct gaccacgatc tgaaacccaa
     3361 tgctgccacg agagatcagt taaatattat tgtgagttat ccaccaacca agcaacttac
     3421 ctatgaagaa caagatcttg tttggaagtt tagatattat cttacgaatc aagaaaaagc
     3481 cttgacaaaa ttcttgaaat gtgttaattg ggatctacct caagaggcca aacaggcctt
     3541 ggaacttctg ggaaaatgga agccgatgga tgtagaggac tccttggagc tgttatcctc
     3601 tcattacacc aacccaactg tgaggcgtta tgctgttgcc cggttgcgac aggccgatga
     3661 tgaggatttg ttgatgtacc tattacaatt ggtccaggct ctcaaatatg aaaattttga
     3721 tgatataaag aatggattgg aacctaccaa gaaggatagt cagagttcag tgtcagaaaa
     3781 tgtgtcaaat tctggaataa attctgcaga aatagatagc tcccaaatta taaccagccc
     3841 ccttccttca gtctcttcac ctcctcctgc atcaaaaaca aaagaagttc cagatggcga
     3901 aaatctggaa caagatctct gtaccttctt gatatcgaga gcctgcaaaa actcaacact
     3961 ggctaattat ttatactggt atgtgatagt ggaatgtgaa gatcaagata ctcagcagag
     4021 agatccaaag acccatgaga tgtacttgaa cgtaatgaga agattcagcc aagcattgtt
     4081 gaagggtgat aagtctgtca gagttatgcg ttctttgctg gctgcacaac agacatttgt
     4141 agatcggttg gtgcatctaa tgaaggcagt acaacgcgaa agtggaaatc gtaagaaaaa
     4201 gaatgagaga ctacaggcat tgcttggaga taatgaaaag atgaatttgt cagatgtgga
     4261 acttatcccg ttgcctttag aaccccaagt gaaaattaga ggaataattc cggaaacagc
     4321 tacactgttt aaaagtgccc ttatgcctgc acagttgttt tttaagacgg aagatggagg
     4381 caaatatcca gttatattta agcatggaga tgatttacgt caagatcaac ttattcttca
     4441 aatcatttca ctcatggaca agctgttacg gaaagaaaat ctggacttga aattgacacc
     4501 ttataaggtg ttagccacca gtacaaaaca tggcttcatg cagtttatcc agtcagttcc
     4561 tgtggctgaa gttcttgata cagagggaag cattcagaac ttttttagaa aatatgcacc
     4621 aagtgagaat gggccaaatg ggattagtgc tgaggtcatg gacacttacg ttaaaagctg
     4681 tgctggatat tgcgtgatca cctatatact tggagttgga gacaggcacc tggataacct
     4741 tttgctaaca aaaacaggca aactcttcca catagacttt ggatatattc tgggtcgtga
     4801 tccaaagcct cttcctccac caatgaagct gaataaagaa atggtagaag gaatgggggg
     4861 cacacagagt gagcagtacc aagagttccg taaacagtgt tacacggctt tcctccacct
     4921 gcgaaggtat tctaatctga ttttgaactt gttttccttg atggttgatg caaacattcc
     4981 agatattgca cttgaaccag ataaaactgt gaaaaaggtt caggataaat tccgcttaga
     5041 cctgtcggat gaagaggctg tgcattacat gcagagtctg attgatgaga gtgtccatgc
     5101 tctttttgct gcagtggtgg aacagattca caagtttgcc cagtactggc gcaaatgagg
     5161 atccagatct aagcttggta ccacgcgtgc gcgctgatcc ggctgctaac aaagcccgaa
     5221 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     5281 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatagccgga taattct
//
