LOCUS       sequence-484467-        3858 bp    DNA     circular SYN 25-AUG-2025
DEFINITION  Bacterial expression of GST-ScAtg38 Delta(182 to 226).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3858)
  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 3858)
  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: pYO1090
FEATURES             Location/Qualifiers
     source          1..3858
                     /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(1659..1677)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1673..1720
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1765..1783)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1851..1955
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1956..2024,2025..2816)
                     /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(2174..2193)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2987..3575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3476..3495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3729..3746
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(3761..3858,1..42)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3826..3845)
                     /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 tcatcagaag aaaataatat tggaatggtc aacaacaata tgcttattgg ctctgttatt
     1321 ttaaacaaca aatctccgat aaatggaata agcaatgcta ggaattggga taacccagca
     1381 tatcaggaca ctttaagtcc cataaacgat ccacttctta tgtcaatttt aaaccgctta
     1441 caatttaatt tgaataatga tattcaattg aaaacagaag gaggaaaaaa ttccaaaaac
     1501 tcggaaatga agattaactt aagactggaa caattcaaaa aggagttggt attatatgag
     1561 cagaaaaaat agggatccag atctaagctt ggtaccacgc gtgcgcgctg atccggctgc
     1621 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
     1681 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatagc
     1741 cggataattc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     1801 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1861 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1921 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     1981 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     2041 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     2101 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     2161 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     2221 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     2281 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     2341 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     2401 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     2461 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     2521 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     2581 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     2641 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     2701 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     2761 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     2821 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     2881 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     2941 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3001 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     3061 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     3121 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     3181 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     3241 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     3301 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     3361 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     3421 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     3481 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     3541 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     3601 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     3661 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     3721 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     3781 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     3841 tgccgcatag ttaagcca
//
