LOCUS       sequence-469204-        2913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Protein expression of His-tagged HsNRBF2 MIT  domain in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2913)
  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 2913)
  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: pOP796
FEATURES             Location/Qualifiers
     source          1..2913
                     /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(714..732)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      728..775
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(820..838)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        906..1010
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1011..1079,1080..1871)
                     /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(1229..1248)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2042..2630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2531..2550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2784..2801
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(2816..2913,1..42)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2881..2900)
                     /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 ccagtaggga tccagatcta agcttggtac cacgcgtgcg
      661 cgctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg ccaccgctga
      721 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
      781 aggaggaact atagccggat aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
      841 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
      901 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      961 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1021 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1081 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1141 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1201 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     1261 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1321 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1381 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1441 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1501 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     1561 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1621 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     1681 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     1741 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     1801 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     1861 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     1921 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     1981 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2041 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2101 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2161 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     2221 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2281 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2341 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2401 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2461 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2521 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2581 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2641 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2701 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     2761 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     2821 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     2881 tacaatctgc tctgatgccg catagttaag cca
//
