LOCUS       sequence-484472-        3549 bp    DNA     circular SYN 25-AUG-2025
DEFINITION  Bacterial expression of GST-ScAtg38 (1-78).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3549)
  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 3549)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 25, 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: pOP775
FEATURES             Location/Qualifiers
     source          1..3549
                     /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             328..981
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     939..961
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1000..1020
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1350..1368)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1364..1411
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1456..1474)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1542..1646
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1647..1715,1716..2507)
                     /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(1865..1884)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2678..3266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3167..3186
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3420..3437
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(3452..3549,1..42)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3517..3536)
                     /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 acataatatg tcccctatac taggttattg gaaaattaag
      361 ggccttgtgc aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat
      421 ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag
      481 tttcccaatc ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc
      541 atacgttata tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag
      601 atttcaatgc ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat
      661 agtaaagact ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa
      721 atgttcgaag atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct
      781 gacttcatgt tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat
      841 gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag
      901 tacttgaaat ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt
      961 ggtggcgacc atcctccaaa atcggattct tctggttccg aaaacctgta tttccagggc
     1021 tcccatatga gtactttagc ggaagtttat actattattg aggacgccga acaggaatgt
     1081 cgaaaaggag attttacaaa tgcaaaagcg aagtatcaag aagccattga agttttaggt
     1141 ccgcagaacg agaatttgtc tcaaaataaa ttaagttctg acgttacgca agccattgac
     1201 ctattaaaac aggacattac agccaaaatc caagaattgg aacttttgat tgaaaagcag
     1261 tagggatcca gatctaagct tggtaccacg cgtgcgcgct gatccggctg ctaacaaagc
     1321 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     1381 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatag ccggataatt
     1441 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1501 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1561 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1621 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1681 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1741 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1801 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1861 ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc
     1921 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     1981 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2041 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2101 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2161 caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat
     2221 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2281 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2341 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2401 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2461 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2521 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2581 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2641 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2701 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2761 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2821 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2881 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2941 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3001 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3061 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3121 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3181 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3241 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3301 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3361 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3421 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     3481 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     3541 gttaagcca
//
