LOCUS       sequence-490366-        6539 bp    DNA     circular SYN 23-OCT-2025
DEFINITION  Expression of ScAtg38 Delta(123 to 226)-3xFLAG under its own 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6539)
  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 6539)
  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: pYO814
FEATURES             Location/Qualifiers
     source          1..6539
                     /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             3428..3493
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3510..3697
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3565..3589)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3766..3786)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3766..3784)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3805..3821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3805..3821)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3818..3840)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3829..3845
                     /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(3853..3859,3860..3877,3878..3883))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3898..3919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4036..4053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4207..4795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4287..4306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4966..5757,5758..5826))
                     /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     5589..5608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5827..5931)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    5968..6471
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6512..6530
                     /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 tgtcgaggac tacaaagacc atgacggtga ttataaagat catgatatcg attacaagga
     3481 tgacgatgac aagtagaatt cgtgagctcg cgaatttctt atgatttatg atttttatta
     3541 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa
     3601 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat
     3661 agcatgaggt cgctcttatt gaccacacct ctaccggcat gccgagcaaa tgcctgcaaa
     3721 tcgctccccg cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt
     3781 aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3841 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3901 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3961 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4021 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4081 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4141 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4201 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4381 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4621 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4741 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4801 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4861 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4921 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4981 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5041 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5101 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5161 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5221 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5281 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     5341 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5401 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5461 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5521 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5581 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5641 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5701 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5761 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5821 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5881 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5941 ccgaaaagtg ccacctgggt ccttttcatc acgtgctata aaaataatta taatttaaat
     6001 tttttaatat aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat
     6061 agtttttgtt ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt
     6121 tatcttgctc ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga
     6181 agaccacaca cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc
     6241 tatttaatct gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat
     6301 tttttaaacc tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt
     6361 actttctaaa atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat
     6421 tattccatca ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtctaaga
     6481 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
