LOCUS       sequence-485582-        6829 bp    DNA     circular SYN 09-SEP-2025
DEFINITION  Expression of ScAtg38 (FL)-3xFLAG under its own promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6829)
  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 6829)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 9, 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: pYO785
FEATURES             Location/Qualifiers
     source          1..6829
                     /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             3740..3805
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3822..4009
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3877..3901)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(4056..4076)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4056..4074)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4095..4111)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4095..4111)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4108..4130)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4119..4135
                     /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(4143..4149,4150..4167,4168..4173))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4188..4209
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4326..4343)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4497..5085)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4577..4596)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5256..6047,6048..6116))
                     /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     5879..5898
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6117..6221)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    6258..6761
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     6802..6820
                     /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 agaaaaaatt
     3601 taaagaatat ggtatgaaaa ttgacgaaat tacgaaagaa aataaaaaat tggctaatga
     3661 aattgggcga ttaagagaac gctgggacag cttagtagaa agtgcgaagc aaagaaggga
     3721 taagcaaaaa aacgtcgagg actacaaaga ccatgacggt gattataaag atcatgatat
     3781 cgattacaag gatgacgatg acaagtagaa ttcgtgagct cgcgaatttc ttatgattta
     3841 tgatttttat tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
     3901 cttaggtttt aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc
     3961 tttctcaggt atagcatgag gtcgctctta ttgaccacac ctctaccggc aggccggccg
     4021 cggccgccac cgcggtggag ctccagcttt tgttcccttt agtgagggtt aattgcgcgc
     4081 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4141 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4201 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4261 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4321 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4381 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4441 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4501 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4561 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4621 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4681 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4741 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4801 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4861 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4921 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4981 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5041 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5101 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5161 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5221 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5281 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5341 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5401 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5461 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5521 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5581 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5641 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5701 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5761 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5821 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5881 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5941 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6001 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6061 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6121 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6181 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6241 ccacctgggt ccttttcatc acgtgctata aaaataatta taatttaaat tttttaatat
     6301 aaatatataa attaaaaata gaaagtaaaa aaagaaatta aagaaaaaat agtttttgtt
     6361 ttccgaagat gtaaaagact ctagggggat cgccaacaaa tactaccttt tatcttgctc
     6421 ttcctgctct caggtattaa tgccgaattg tttcatcttg tctgtgtaga agaccacaca
     6481 cgaaaatcct gtgattttac attttactta tcgttaatcg aatgtatatc tatttaatct
     6541 gcttttcttg tctaataaat atatatgtaa agtacgcttt ttgttgaaat tttttaaacc
     6601 tttgtttatt tttttttctt cattccgtaa ctcttctacc ttctttattt actttctaaa
     6661 atccaaatac aaaacataaa aataaataaa cacagagtaa attcccaaat tattccatca
     6721 ttaaaagata cgaggcgcgt gtaagttaca ggcaagcgat ccgtctaaga aaccattatt
     6781 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtc
//
