LOCUS       sequence-490368-        6674 bp    DNA     circular SYN 23-OCT-2025
DEFINITION  Expression of ScAtg38 Delta(123 to 181)-3xFLAG under its own 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6674)
  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 6674)
  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: pYO827
FEATURES             Location/Qualifiers
     source          1..6674
                     /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             3563..3628
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3645..3832
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3700..3724)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(3901..3921)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3901..3919)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3940..3956)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3940..3956)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3953..3975)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3964..3980
                     /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(3988..3994,3995..4012,4013..4018))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4033..4054
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4171..4188)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4342..4930)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4422..4441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5101..5892,5893..5961))
                     /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     5724..5743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5962..6066)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6103..6606
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6647..6665
                     /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 ttttaaagaa tatggtatga aaattgacga aattacgaaa gaaaataaaa aattggctaa
     3481 tgaaattggg cgattaagag aacgctggga cagcttagta gaaagtgcga agcaaagaag
     3541 ggataagcaa aaaaacgtcg aggactacaa agaccatgac ggtgattata aagatcatga
     3601 tatcgattac aaggatgacg atgacaagta gaattcgtga gctcgcgaat ttcttatgat
     3661 ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg
     3721 actcttaggt tttaaaacga aaattcttat tcttgagtaa ctctttcctg taggtcaggt
     3781 tgctttctca ggtatagcat gaggtcgctc ttattgacca cacctctacc ggcatgccga
     3841 gcaaatgcct gcaaatcgct ccccgcggcc gccaccgcgg tggagctcca gcttttgttc
     3901 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3961 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     4021 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     4081 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     4141 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4201 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4261 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4321 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4381 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4441 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gacacctgtc
     4501 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4561 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4621 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4681 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4741 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4801 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4861 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4921 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4981 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5041 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5101 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5161 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5221 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5281 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5341 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5401 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5461 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5521 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5581 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5641 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5701 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5761 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5821 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5881 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5941 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6001 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6061 tccgcgcaca tttccccgaa aagtgccacc tgggtccttt tcatcacgtg ctataaaaat
     6121 aattataatt taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga
     6181 aattaaagaa aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca
     6241 acaaatacta ccttttatct tgctcttcct gctctcaggt attaatgccg aattgtttca
     6301 tcttgtctgt gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt
     6361 aatcgaatgt atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac
     6421 gctttttgtt gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt
     6481 ctaccttctt tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag
     6541 agtaaattcc caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa
     6601 gcgatccgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     6661 gaggcccttt cgtc
//
