LOCUS       sequence-470270-        3537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression His-taged HsNRBF2_FL in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3537)
  AUTHORS   Ohashi Y, Soler N, Garcia Ortegon M, Zhang L, Kirsten ML, Perisic O,
            Masson GR, Burke JE, Jakobi AJ, Apostolakis AA, Johnson CM, Ohashi 
            M, Ktistakis NT, Sachse C, Williams RL
  TITLE     Characterization of Atg38 and NRBF2, a fifth subunit of the 
            autophagic Vps34/PIK3C3 complex.
  JOURNAL   Autophagy. 2016 Nov;12(11):2129-2144. doi: 
            10.1080/15548627.2016.1226736. Epub 2016 Sep 14.
  PUBMED    27630019
REFERENCE   2  (bases 1 to 3537)
  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: 
            "10.1080/15548627.2016.1226736"; journalName: "Autophagy"; date: 
            "2016-11"; volume: "12"; issue: "11"; pages: "2129-2144"
COMMENT     Sequence Label: pOP795
FEATURES             Location/Qualifiers
     source          1..3537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..150
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     203..222
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        203..221
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             295..317
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             361..381
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1338..1356)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1352..1399
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1444..1462)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1530..1634
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1635..1703,1704..2495)
                     /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(1853..1872)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2666..3254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3155..3174
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3408..3425
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(3440..3537,1..42)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3505..3524)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
       61 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
      121 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
      181 caggatctcg atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc
      241 tctagtgccg gctccggaga gctccaattg gaattcgcta gctctagaaa taattttgtt
      301 taactttaag aaggagatat aaatatggct caccatcatc atcatcattc ttctggttcc
      361 gaaaacctgt atttccaggg ctcccatatg gaagtaatgg aaggacccct caacctggct
      421 catcaacaga gcagacgagc agaccgttta ttagctgcag gcaaatacga agaggctatt
      481 tcttgtcaca aaaaggctgc agcatatctt tctgaagcca tgaagctgac acagtcagag
      541 caggctcatc tttcactgga attgcaaagg gatagccata tgaaacagct cctcctcatc
      601 caagagagat ggaaaagggc ccagcgtgaa gaaagattga aagcccagca gaacacagac
      661 aaggatgcag ctgcccatct tcagacatct cacaaaccct ctgcagagga tgcagagggc
      721 cagagtcccc tttctcagaa gtacagccct tccacagaga aatgcctgcc tgagattcag
      781 gggatctttg acagggatcc agacacacta ctttatttac ttcagcaaaa gagtgagcca
      841 gcagagccat gtattggaag caaagcccca aaagatgata aaacaattat agaggagcag
      901 gcaaccaaaa ttgcagattt gaagaggcat gtggaattcc ttgtggctga gaatgaaaga
      961 ttaaggaaag aaaataaaca actaaaggct gaaaaggcca gacttctaaa aggtccaata
     1021 gaaaaggagc tggatgtaga tgctgatttt gtagaaacgt cagagttatg gagcttgcca
     1081 ccacatgcag aaactgctac agcctcctca acctggcaga agttcgcagc aaatactggg
     1141 aaagccaagg acattccaat ccccaatctt cctcccttgg attttccatc tccagaactt
     1201 cctcttatgg agctctctga ggatattctg aaaggattta tgaataatta gggatccaga
     1261 tctaagcttg gtaccacgcg tgcgcgctga tccggctgct aacaaagccc gaaaggaagc
     1321 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     1381 ggtcttgagg ggttttttgc tgaaaggagg aactatagcc ggataattct tgaagacgaa
     1441 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     1501 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     1561 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     1621 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     1681 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     1741 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     1801 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     1861 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     1921 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     1981 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     2041 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     2101 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     2161 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     2221 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     2281 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     2341 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     2401 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     2461 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     2521 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     2581 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     2641 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2701 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2761 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     2821 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     2881 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     2941 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3001 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3061 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3121 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3181 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3241 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3301 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3361 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3421 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     3481 cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagcca
//
