LOCUS       sequence-329318-        9204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Atg1 with ATG1 3'UTR in yeasts..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9204)
  AUTHORS   Gatica D, Hu G, Liu X, Zhang N, Williamson PR, Klionsky DJ
  TITLE     The Pat1-Lsm Complex Stabilizes ATG mRNA during Nitrogen 
            Starvation-Induced Autophagy.
  JOURNAL   Mol Cell. 2019 Jan 17;73(2):314-324.e4. doi: 
            10.1016/j.molcel.2018.11.002. Epub 2018 Dec 6.
  PUBMED    30527663
REFERENCE   2  (bases 1 to 9204)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.molcel.2018.11.002"; journalName: "Mol Cell"; date: 
            "2019-01-17- 17"; volume: "73"; issue: "2"; pages: "314-324.e4"
COMMENT     Sequence Label: pRS406 ATG1
FEATURES             Location/Qualifiers
     source          1..9204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    94..115
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(130..135,136..153,154..160)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    168..184
                     /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)."
     primer_bind     173..195
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     192..208
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     192..208
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     227..247
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        229..247
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     284..300
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     284..300
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5206..5225)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5207..5225)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5235..5252)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5235..5251)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5244..5266)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5392..5847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5479..5498)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5689..5710
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5978..6781)
                     /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"
     promoter        complement(6782..6997)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7021..7040)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7140..7162
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7200..7218)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7286..7390
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7391..7459,7460..8251)
                     /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     complement(7609..7628)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8422..9010
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8911..8930
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9164..9181
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
       61 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
      121 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
      181 acaatttcac acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca
      241 ctaaagggaa caaaagctgg agctccaccg cggtggcggc cgctctagaa ctagtggatc
      301 ccccgggacc tgccacaagg ttatttctac acacaataaa aataagatac attgccgctg
      361 ctccgcatta catatcatat agagtgacac taagtaatta aatgacacta agtaacgaaa
      421 gatatctgta aacaacgact atataccaga attagtcata ttctctattg tattcaccct
      481 acacagccgc attacacagc tgctccggac gttttacgct gcacatcaat gactaatacc
      541 actgtatttg atccacactt agcgcaaaag tcacctgggt ggcgttttcc gctcgcgctc
      601 cgccggtgag agcgagaaaa aaaaaaggga ttgggcggta atgtaaggaa aacccactag
      661 ttaaaatcaa tgagataagg aactctcttt ttttcccgtt aatcatcttt tccttgttcg
      721 tttcgtgtat ctgtaatgaa tcttatcgga gtaactatta ctggaaagcc ataccgttaa
      781 agaagcaagc atatcattcg tattcaagca tctgtttttc tttttaaccg ctcggctctg
      841 atttcttaaa ccatttcccc catcagcatc agtttgtgat ttgtaactat atcgatcgtg
      901 gaatcgatgg aattctaagc gtcatttagt tggcaacaaa aggaatcgag catctcataa
      961 aggagcttat attattaata tcttcctgat cgtactattt ttttttagct ttttattttt
     1021 taggccgagg ttaattctag aacgccacat ttcatcacct ttaagttaag taccaaggcc
     1081 atcttttacc ctttttagtt tctggggaaa cagagaacag tactatagaa gataaaaaga
     1141 tataatacaa tcgtagtaat agtacccata ataaaataac atataatata ataaaaacat
     1201 aaggcaaagg agataggaga ataatcattg atatctccac aaaacgaatt tgaagctacc
     1261 ccatattttc aaatctcttt tacaacacca gacgagaaat taagaaactc gagatgggag
     1321 acattaaaaa taaagatcac acaacctctg tgaaccataa tctaatggca agtgcaggaa
     1381 attacaccgc tgagaaagaa atcggaaagg gttcgtttgc cactgtatat agagggcatc
     1441 tgacatccga caaatctcag catgtagcca taaaggaagt atcaagggcg aaattaaaaa
     1501 ataagaaatt actagagaat ttggaaatag aaatcgctat cttgaagaaa atcaagcatc
     1561 ctcatatcgt cggacttatt gactgtgaac gaacatcaac agatttttat ttgatcatgg
     1621 agtactgtgc tcttggggac ctaacatttc tgttgaaaag gcgtaaagaa ttgatggaga
     1681 atcatcctct actaaggacc gtatttgaaa aataccctcc accgagtgag aaccataacg
     1741 gcctgcatag agcgtttgtc ttgagttatt tacagcagtt agcgtccgct ttgaaatttt
     1801 tgaggtctaa aaacttggtt catagagaca tcaagcctca aaacttacta ttatctacac
     1861 ccctaattgg atatcatgat tcaaaaagct tccatgaact tggatttgtt gggatctaca
     1921 acttacccat tttaaagata gcagatttcg ggtttgcaag atttttgcca aacacgtcat
     1981 tagcagaaac tctttgtggc tcaccattat atatggcacc agaaattttg aattatcaaa
     2041 aatataacgc taaagcagat ctgtggtctg tcggtacagt ggtattcgaa atgtgctgtg
     2101 gcaccccacc gtttagagct tccaatcatt tggagttatt caagaaaatt aaaagagcaa
     2161 acgatgtcat aacgtttcct tcatattgca atattgaacc agagttaaaa gagttgatat
     2221 gtagtttatt gacatttgat ccagcccaaa gaataggatt tgaggagttt tttgctaaca
     2281 aggtagtcaa cgaagacttg tcttcttatg aattggaaga tgatttacct gagttagaat
     2341 ccaaatcaaa aggtattgta gaaagtaata tgttcgtttc tgagtattta tctaaacagc
     2401 caaagagtcc gaacagtaat cttgcaggtc atcaatcaat ggcagataat cccgcggagc
     2461 tcagtgatgc cctcaagaac agcaatatat taactgcccc agccgttaaa acagaccata
     2521 cacaagccgt agataaaaag gcttcaaata ataaatacca taatagccta gtttcagata
     2581 gaagctttga aagagaatat gtggtggtag agaagaaatc ggttgaagtt aattcattgg
     2641 cagacgaggt tgctcaagca ggattcaatc caaatcctat taagcaccca acttcaactc
     2701 aaaatcagaa cgtcttattg aatgagcaat tctctccaaa caatcaacag tattttcaaa
     2761 atcaaggaga aaatccgagg ttactgaggg ccacatcatc ttccagtgga ggtagtgatg
     2821 ggtctaggcg accatctttg gtggatagac gcttgtctat atcctcgctg aatccatcta
     2881 atgcattatc aagagccctc ggtattgcat caacgagatt gtttggtggt gcaaatcaac
     2941 agcagcaaca gcaacaaatc acatcttccc caccgtacag tcaaactttg ttgaattccc
     3001 aactttttca tgaacttact gagaatataa tattaagaat agatcaccta cagcatccag
     3061 agacactgaa attagataat actaatatcg ttagtatttt ggaatctctg gccgcaaagg
     3121 catttgttgt ttactcttat gcagaagtga aattttctca aattgttcca ttatcaacaa
     3181 cattaaaagg catggctaac tttgagaaca ggcgcagtat ggatagtaat gctattgcag
     3241 aagaacaaga ctcagacgat gcagaagaag aggatgaaac gttgaaaaag tacaaggaag
     3301 attgcttatc cacgaaaact tttggaaagg gtagaacttt atctgccaca tctcagttga
     3361 gtgcaacttt caataaacta ccacgttcgg aaatgatcct tctatgtaat gaggccattg
     3421 tcttatatat gaaggcatta tccattttat caaaatctat gcaggtaacg tccaactggt
     3481 ggtatgaatc tcaagaaaaa tcatgttctc taagagttaa cgtgttggta cagtggctaa
     3541 gggaaaaatt taatgaatgt ttagaaaaag ctgatttcct gagattaaaa attaacgact
     3601 tgagattcaa gcatgcttct gaggtagctg aaaatcaaac tttggaagaa aaaggtagtt
     3661 cggaagagcc agtatattta gaaaagctat tatatgatcg tgcattagaa atatctaaga
     3721 tggccgcaca tatggaatta aaaggagaaa atttgtacaa ctgtgaatta gcttatgcaa
     3781 cttcactttg gatgttagaa acttctctgg acgatgatga ttttacgaat gcctatggtg
     3841 actacccttt taaaactaac atacatctaa aaagtaacga cgttgaagat aaagagaagt
     3901 atcatagtgt gttggatgaa aatgacagaa taatcataag aaagtatatt gatagtattg
     3961 caaacaggtt gaaaatattg aggcagaaga tgaaccacca aaattaagtg atgcataata
     4021 tggttttctt attaagtaca aatgacctgc tatattttca agtatcttca tgattatgtt
     4081 atgctactct cgaaatgctt tacgagttcg tcattaaagt attcaaattt tattttccct
     4141 ttgtgttttg ttttgttttt atttctacgt gatttttttc ttgaaatcca gctcttttat
     4201 acctggatgg gttttatttt gatgtttaaa tgttaattaa ggaaattatt gaattccttc
     4261 ggtttcggtg tgtttttttc tttttaagtt tttttttgtt gcgtttagta cttgcctttg
     4321 gctatacata tccttttaat ctaattttta tttcttcaaa atttgattct ttaacctttt
     4381 gttctatata tatatataaa tgtatgtatg tatgatttga ttgaggaggc tttaacaagt
     4441 cattaatatc tttattattt ttacttgttg agttcattaa aattcatttt ataatttgtt
     4501 gttacaaaca gtattttaca agtcataata aacgttcaaa aacaaacaca atattaaaag
     4561 aattggtatt aggatccgac atgacaatta acttagatcc ttctatcaag tttctaatca
     4621 tatattacat ctattaaaat aatttacata tatcatgcta aagcttataa agagacattc
     4681 ttcttctcgt gcgctgggcg tcgctagcct caaatacaag tgatgcgtct gcttctattc
     4741 taaggtcatc cttgctgtta gatttccctt ctaccaggtg ggcttttata cgttgaggta
     4801 cattaccgac aggcgtaaga gtagagctca aacttgcttc cgtactctca aagggatatt
     4861 ctcctacagg agacatagtt gtatattgtg gaaactcaca agaattcact cttaatgaac
     4921 ttccactctc acctatcaag cattgttgat caccgattgc tggggcatcc tgaatactga
     4981 tctgagatgc aaaagcggtg gatgatgggt accgtactgg tcatacaaag ctcttaaacg
     5041 attgtcttta agtatttcgt atgcctggca tatctcttga aactttcgct tggattcttc
     5101 tgtatgattg tttttatcag gatgatattt taatgcagcg ttcctgtaag ctttcttaat
     5161 ctcagatgtg gatgcaaaag cggtggatga tgggtaccca attcgcccta tagtgagtcg
     5221 tattacgcgc gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     5281 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     5341 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     5401 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     5461 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     5521 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     5581 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     5641 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5701 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     5761 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5821 ttaacaaaat attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg
     5881 cggtatttca caccgcatag ggtaataact gatataatta aattgaagct ctaatttgtg
     5941 agtttagtat acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc
     6001 tcaaatatgc ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc
     6061 cctttgcaaa tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat
     6121 ccacggttct atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg
     6181 tcataatcaa ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc
     6241 cgataacaaa atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag
     6301 tagataggga gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg
     6361 ttacttcttc tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg
     6421 cattcgtaat gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga
     6481 ctgtattacc aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg
     6541 ataatgcctt tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat
     6601 gtgtttttag taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg
     6661 tggtacgaac atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata
     6721 gcttggcagc aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca
     6781 tgatttatct tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc
     6841 aatttcatgt ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct
     6901 tccttcgttc ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa
     6961 atcaaaaaaa agaataaaaa aaaaatgatg aattgaaaag gtggtatggt gcactctcag
     7021 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     7081 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     7141 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     7201 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     7261 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     7321 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     7381 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     7441 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     7501 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     7561 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     7621 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     7681 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     7741 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     7801 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     7861 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     7921 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     7981 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     8041 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     8101 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     8161 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     8221 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     8281 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     8341 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     8401 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     8461 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     8521 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     8581 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     8641 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     8701 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     8761 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     8821 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     8881 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     8941 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     9001 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     9061 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     9121 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     9181 ggaagcggaa gagcgcccaa tacg
//
