LOCUS       sequence-490367-        6716 bp    DNA     circular SYN 23-OCT-2025
DEFINITION  Expression of ScAtg38 Delta(182 to 226)-3xFLAG under its own 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6716)
  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 6716)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 23, 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: pYO815
FEATURES             Location/Qualifiers
     source          1..6716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        194..409
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             410..1213
                     /codon_start=1
                     /gene="S. cerevisiae URA3"
                     /product="orotidine-5'-phosphate decarboxylase, required 
                     for uracil biosynthesis"
                     /label=URA3
                     /note="yeast auxotrophic marker, counterselectable with 
                     5-fluoroorotic acid (5-FOA)"
                     /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
                     ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
                     YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
                     YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
                     VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
     rep_origin      complement(1344..1799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1481..1502)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1693..1712
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1925..1947
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1939..1956
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1940..1956
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1966..1985
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1966..1984
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3605..3670
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3687..3874
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3742..3766)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3943..3963)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3943..3961)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3982..3998)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3982..3998)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3995..4017)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4006..4022
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4030..4036,4037..4054,4055..4060))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4075..4096
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4213..4230)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4384..4972)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4464..4483)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5143..5934,5935..6003))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5766..5785
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6004..6108)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6145..6648
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6689..6707
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc
      241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag
      301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt
      361 cttaacccaa ctgcacagaa caaaaacctg caggaaacga agataaatca tgtcgaaagc
      421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat
      481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt
      541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat
      601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta
      661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca
      721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt
      781 ggtgggccca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc
      841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata
      901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc
      961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt
     1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt
     1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc
     1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg
     1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag
     1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg
     1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt
     1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     1921 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta
     1981 tagggcgaat tgggtaccat gtccagtatt tactctacct tgattccttc tcctgaactt
     2041 catttcctaa gccctagttt cactccattt acatcagatt atattcacga tgatatagcg
     2101 cataaaggcc attccagtta tgacgtcatg ttagatttat tagacccttc caattctctt
     2161 gtctctaccg cgatgaacaa tccaacatat tttaacgttt ataatacaat cattggtaat
     2221 gtagaacctc gtcagatatc gcgtgctatg accaaattac aacaacgcat caaattcccc
     2281 tcatggtcct cttctgcgat gcacgttaac attggaagaa gatcgccata cttgccttta
     2341 caaccaaatg aaaacgaagt tagcggcatg atgttaagca atatgtctac cgtggtgaac
     2401 gtctttgaga atgcgtgcaa tacttttgac aaagtatttg ccaagggcgc atttttaaat
     2461 aattataatg taggcgactt gttccaatca atgcagaacg ttcaagatga attcgccgag
     2521 tcaagagaag tagtacaaag cctgatggag gattatgtag ctgcagaaca agattcgtac
     2581 ttggacgatg tacttgttga tgatgaaaat atggttggcg agttggaaga ggacctggat
     2641 gccgacggtg atcataaatt agtataatga tgccttcctt gttcaggaat tttaccaccg
     2701 cccaatagtt ttattatagt actctcaata tatctatacc aacaaccgct tcaaatgtat
     2761 tatctccctt tataaattct gcactgtatt gtaatatgta taaagctctg taccactttt
     2821 tcatcattgc atacatcgat agcgcctctt caactttaga atttcctaaa gggtagaaaa
     2881 accagatgat tcaactcagc gtaatggaac gctctgtttg aaataaaaaa catctgtatc
     2941 gctgcacgga ttagttcgat aattcaacga cagttatact aatcttggtg atggaatgag
     3001 tactttagcg gaagtaagtt tttgaattat gcccccactt ttttttttgt tcatggtgac
     3061 taacatgaat taggtttata ctattattga ggacgccgaa caggaatgtc gaaaaggaga
     3121 ttttacaaat gcaaaagcga agtatcaaga agccattgaa gttttaggtc cgcagaacga
     3181 gaatttgtct caaaataaat taagttctga cgttacgcaa gccattgacc tattaaaaca
     3241 ggacattaca gccaaaatcc aagaattgga acttttgatt gaaaagcagt catcagaaga
     3301 aaataatatt ggaatggtca acaacaatat gcttattggc tctgttattt taaacaacaa
     3361 atctccgata aatggaataa gcaatgctag gaattgggat aacccagcat atcaggacac
     3421 tttaagtccc ataaacgatc cacttcttat gtcaatttta aaccgcttac aatttaattt
     3481 gaataatgat attcaattga aaacagaagg aggaaaaaat tccaaaaact cggaaatgaa
     3541 gattaactta agactggaac aattcaaaaa ggagttggta ttatatgagc agaaaaaagt
     3601 cgaggactac aaagaccatg acggtgatta taaagatcat gatatcgatt acaaggatga
     3661 cgatgacaag tagaattcgt gagctcgcga atttcttatg atttatgatt tttattatta
     3721 aataagttat aaaaaaaata agtgtataca aattttaaag tgactcttag gttttaaaac
     3781 gaaaattctt attcttgagt aactctttcc tgtaggtcag gttgctttct caggtatagc
     3841 atgaggtcgc tcttattgac cacacctcta ccggcatgcc gagcaaatgc ctgcaaatcg
     3901 ctccccgcgg ccgccaccgc ggtggagctc cagcttttgt tccctttagt gagggttaat
     3961 tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4021 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4081 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     4141 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     4201 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4261 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4321 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4381 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4441 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4501 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4561 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4621 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4681 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4741 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4801 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4861 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4921 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4981 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5041 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5101 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     5161 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     5221 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     5281 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     5341 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     5401 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     5461 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     5521 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     5581 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5641 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5701 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5761 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5821 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5881 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5941 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6001 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6061 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6121 aaaagtgcca cctgggtcct tttcatcacg tgctataaaa ataattataa tttaaatttt
     6181 ttaatataaa tatataaatt aaaaatagaa agtaaaaaaa gaaattaaag aaaaaatagt
     6241 ttttgttttc cgaagatgta aaagactcta gggggatcgc caacaaatac taccttttat
     6301 cttgctcttc ctgctctcag gtattaatgc cgaattgttt catcttgtct gtgtagaaga
     6361 ccacacacga aaatcctgtg attttacatt ttacttatcg ttaatcgaat gtatatctat
     6421 ttaatctgct tttcttgtct aataaatata tatgtaaagt acgctttttg ttgaaatttt
     6481 ttaaaccttt gtttattttt ttttcttcat tccgtaactc ttctaccttc tttatttact
     6541 ttctaaaatc caaatacaaa acataaaaat aaataaacac agagtaaatt cccaaattat
     6601 tccatcatta aaagatacga ggcgcgtgta agttacaggc aagcgatccg tctaagaaac
     6661 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtc
//
