LOCUS       sequence-329347-        8734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Atg1 with ATG17 3'UTR in yeasts..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8734)
  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 8734)
  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-ATG17-3'UTR
FEATURES             Location/Qualifiers
     source          1..8734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     653..673
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        655..673
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     710..726
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     710..726
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5162..5181)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5163..5181)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5191..5208)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5191..5207)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5200..5222)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5348..5803
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5435..5454)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5645..5666
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5934..6737)
                     /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(6738..6953)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6977..6996)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7096..7118
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7156..7174)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7242..7346
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7347..7415,7416..8207)
                     /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(7565..7584)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(8378..8734,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
      661 ccctcactaa agggaacaaa agctggagct ccaccgcggt ggcggccgct ctagaactag
      721 tggatccccc gggacctgcc acaaggttat ttctacacac aataaaaata agatacattg
      781 ccgctgctcc gcattacata tcatatagag tgacactaag taattaaatg acactaagta
      841 acgaaagata tctgtaaaca acgactatat accagaatta gtcatattct ctattgtatt
      901 caccctacac agccgcatta cacagctgct ccggacgttt tacgctgcac atcaatgact
      961 aataccactg tatttgatcc acacttagcg caaaagtcac ctgggtggcg ttttccgctc
     1021 gcgctccgcc ggtgagagcg agaaaaaaaa aagggattgg gcggtaatgt aaggaaaacc
     1081 cactagttaa aatcaatgag ataaggaact ctcttttttt cccgttaatc atcttttcct
     1141 tgttcgtttc gtgtatctgt aatgaatctt atcggagtaa ctattactgg aaagccatac
     1201 cgttaaagaa gcaagcatat cattcgtatt caagcatctg tttttctttt taaccgctcg
     1261 gctctgattt cttaaaccat ttcccccatc agcatcagtt tgtgatttgt aactatatcg
     1321 atcgtggaat cgatggaatt ctaagcgtca tttagttggc aacaaaagga atcgagcatc
     1381 tcataaagga gcttatatta ttaatatctt cctgatcgta ctattttttt ttagcttttt
     1441 attttttagg ccgaggttaa ttctagaacg ccacatttca tcacctttaa gttaagtacc
     1501 aaggccatct tttacccttt ttagtttctg gggaaacaga gaacagtact atagaagata
     1561 aaaagatata atacaatcgt agtaatagta cccataataa aataacatat aatataataa
     1621 aaacataagg caaaggagat aggagaataa tcattgatat ctccacaaaa cgaatttgaa
     1681 gctaccccat attttcaaat ctcttttaca acaccagacg agaaattaag aaactcgaga
     1741 tgggagacat taaaaataaa gatcacacaa cctctgtgaa ccataatcta atggcaagtg
     1801 caggaaatta caccgctgag aaagaaatcg gaaagggttc gtttgccact gtatatagag
     1861 ggcatctgac atccgacaaa tctcagcatg tagccataaa ggaagtatca agggcgaaat
     1921 taaaaaataa gaaattacta gagaatttgg aaatagaaat cgctatcttg aagaaaatca
     1981 agcatcctca tatcgtcgga cttattgact gtgaacgaac atcaacagat ttttatttga
     2041 tcatggagta ctgtgctctt ggggacctaa catttctgtt gaaaaggcgt aaagaattga
     2101 tggagaatca tcctctacta aggaccgtat ttgaaaaata ccctccaccg agtgagaacc
     2161 ataacggcct gcatagagcg tttgtcttga gttatttaca gcagttagcg tccgctttga
     2221 aatttttgag gtctaaaaac ttggttcata gagacatcaa gcctcaaaac ttactattat
     2281 ctacacccct aattggatat catgattcaa aaagcttcca tgaacttgga tttgttggga
     2341 tctacaactt acccatttta aagatagcag atttcgggtt tgcaagattt ttgccaaaca
     2401 cgtcattagc agaaactctt tgtggctcac cattatatat ggcaccagaa attttgaatt
     2461 atcaaaaata taacgctaaa gcagatctgt ggtctgtcgg tacagtggta ttcgaaatgt
     2521 gctgtggcac cccaccgttt agagcttcca atcatttgga gttattcaag aaaattaaaa
     2581 gagcaaacga tgtcataacg tttccttcat attgcaatat tgaaccagag ttaaaagagt
     2641 tgatatgtag tttattgaca tttgatccag cccaaagaat aggatttgag gagttttttg
     2701 ctaacaaggt agtcaacgaa gacttgtctt cttatgaatt ggaagatgat ttacctgagt
     2761 tagaatccaa atcaaaaggt attgtagaaa gtaatatgtt cgtttctgag tatttatcta
     2821 aacagccaaa gagtccgaac agtaatcttg caggtcatca atcaatggca gataatcccg
     2881 cggagctcag tgatgccctc aagaacagca atatattaac tgccccagcc gttaaaacag
     2941 accatacaca agccgtagat aaaaaggctt caaataataa ataccataat agcctagttt
     3001 cagatagaag ctttgaaaga gaatatgtgg tggtagagaa gaaatcggtt gaagttaatt
     3061 cattggcaga cgaggttgct caagcaggat tcaatccaaa tcctattaag cacccaactt
     3121 caactcaaaa tcagaacgtc ttattgaatg agcaattctc tccaaacaat caacagtatt
     3181 ttcaaaatca aggagaaaat ccgaggttac tgagggccac atcatcttcc agtggaggta
     3241 gtgatgggtc taggcgacca tctttggtgg atagacgctt gtctatatcc tcgctgaatc
     3301 catctaatgc attatcaaga gccctcggta ttgcatcaac gagattgttt ggtggtgcaa
     3361 atcaacagca gcaacagcaa caaatcacat cttccccacc gtacagtcaa actttgttga
     3421 attcccaact ttttcatgaa cttactgaga atataatatt aagaatagat cacctacagc
     3481 atccagagac actgaaatta gataatacta atatcgttag tattttggaa tctctggccg
     3541 caaaggcatt tgttgtttac tcttatgcag aagtgaaatt ttctcaaatt gttccattat
     3601 caacaacatt aaaaggcatg gctaactttg agaacaggcg cagtatggat agtaatgcta
     3661 ttgcagaaga acaagactca gacgatgcag aagaagagga tgaaacgttg aaaaagtaca
     3721 aggaagattg cttatccacg aaaacttttg gaaagggtag aactttatct gccacatctc
     3781 agttgagtgc aactttcaat aaactaccac gttcggaaat gatccttcta tgtaatgagg
     3841 ccattgtctt atatatgaag gcattatcca ttttatcaaa atctatgcag gtaacgtcca
     3901 actggtggta tgaatctcaa gaaaaatcat gttctctaag agttaacgtg ttggtacagt
     3961 ggctaaggga aaaatttaat gaatgtttag aaaaagctga tttcctgaga ttaaaaatta
     4021 acgacttgag attcaagcat gcttctgagg tagctgaaaa tcaaactttg gaagaaaaag
     4081 gtagttcgga agagccagta tatttagaaa agctattata tgatcgtgca ttagaaatat
     4141 ctaagatggc cgcacatatg gaattaaaag gagaaaattt gtacaactgt gaattagctt
     4201 atgcaacttc actttggatg ttagaaactt ctctggacga tgatgatttt acgaatgcct
     4261 atggtgacta cccttttaaa actaacatac atctaaaaag taacgacgtt gaagataaag
     4321 agaagtatca tagtgtgttg gatgaaaatg acagaataat cataagaaag tatattgata
     4381 gtattgcaaa caggttgaaa atattgaggc agaagatgaa ccaccaaaat taaaaaaaag
     4441 gaaaaaaagg atacggtaca aagattcaat aattgtatca atatttcgaa aaatacttta
     4501 tcaaaaaagc aatacttgaa ggagaaagga aaaaattcgc tgtcttcttc acttttacga
     4561 caaagaaaaa tgtgcccttt tcaactaaaa gtcatgaatg aataaataaa tgattacata
     4621 tgcgtatatt ttatagtgaa caacttctga catattatcc cgaatgtgag taatcatgct
     4681 tgtgctgtga atctgtgttg gtgtttggct gtgaccctgc caaaaaggtg ctcgaagtgc
     4741 cggaaacaat tccactttta tccaaactgc tcgtagtgga cgagtgggat aacgttcttg
     4801 ttaaatattt tccaaaaagt aaactgtttg ggccaaatga gtcgctatgt tctaatacaa
     4861 attggctatg attggaccct gaggtgtgat gatcccttcc agaaaagctt gcctttttat
     4921 tggggttctt gtgtgatctg agatgcaaaa gcggtggatg atgggtaccg tactggtcat
     4981 acaaagctct taaacgattg tctttaagta tttcgtatgc ctggcatatc tcttgaaact
     5041 ttcgcttgga ttcttctgta tgattgtttt tatcaggatg atattttaat gcagcgttcc
     5101 tgtaagcttt cttaatctca gatgtggatg caaaagcggt ggatgatggg tacccaattc
     5161 gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg
     5221 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     5281 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     5341 cgaatggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     5401 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     5461 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     5521 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     5581 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5641 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     5701 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     5761 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc
     5821 tccttacgca tctgtgcggt atttcacacc gcatagggta ataactgata taattaaatt
     5881 gaagctctaa tttgtgagtt tagtatacat gcatttactt ataatacagt tttttagttt
     5941 tgctggccgc atcttctcaa atatgcttcc cagcctgctt ttctgtaacg ttcaccctct
     6001 accttagcat cccttccctt tgcaaatagt cctcttccaa caataataat gtcagatcct
     6061 gtagagacca catcatccac ggttctatac tgttgaccca atgcgtctcc cttgtcatct
     6121 aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt catctcttcc acccatgtct
     6181 ctttgagcaa taaagccgat aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc
     6241 ttagtatatt ctccagtaga tagggagccc ttgcatgaca attctgctaa catcaaaagg
     6301 cctctaggtt cctttgttac ttcttctgcc gcctgcttca aaccgctaac aatacctggg
     6361 cccaccacac cgtgtgcatt cgtaatgtct gcccattctg ctattctgta tacacccgca
     6421 gagtactgca atttgactgt attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa
     6481 aaattgtact tggcggataa tgcctttagc ggcttaactg tgccctccat ggaaaaatca
     6541 gtcaagatat ccacatgtgt ttttagtaaa caaattttgg gacctaatgc ttcaactaac
     6601 tccagtaatt ccttggtggt acgaacatcc aatgaagcac acaagtttgt ttgcttttcg
     6661 tgcatgatat taaatagctt ggcagcaaca ggactaggat gagtagcagc acgttcctta
     6721 tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg
     6781 gttaagaata ctgggcaatt tcatgtttct tcaacactac atatgcgtat atataccaat
     6841 ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa
     6901 tttcaaggaa accgaaatca aaaaaaagaa taaaaaaaaa atgatgaatt gaaaaggtgg
     6961 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     7021 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     7081 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     7141 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     7201 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     7261 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     7321 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     7381 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     7441 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     7501 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     7561 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     7621 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     7681 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     7741 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     7801 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     7861 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     7921 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     7981 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     8041 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     8101 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     8161 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     8221 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     8281 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     8341 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     8401 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     8461 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     8521 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     8581 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     8641 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     8701 ggggttcgtg cacacagccc agcttggagc gaac
//
