LOCUS       sequence-329346-        8549 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Atg1 with ATG7 3'UTR in yeasts..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8549)
  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 8549)
  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-ATG7-3'UTR
FEATURES             Location/Qualifiers
     source          1..8549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    75..96
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(111..116,117..134,135..141)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    149..165
                     /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     154..176
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     173..189
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     173..189
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     208..228
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        210..228
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     265..281
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     265..281
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4532..4551)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4533..4551)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4561..4578)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4561..4577)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4570..4592)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4718..5173
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4805..4824)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5015..5036
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5304..6107)
                     /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(6108..6323)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6347..6366)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6466..6488
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6526..6544)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6612..6716
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6717..6785,6786..7577)
                     /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(6935..6954)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7748..8336
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8237..8256
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8490..8507
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
       61 gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc tttacacttt
      121 atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac
      181 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg
      241 gagctccacc gcggtggcgg ccgctctaga actagtggat cccccgggac ctgccacaag
      301 gttatttcta cacacaataa aaataagata cattgccgct gctccgcatt acatatcata
      361 tagagtgaca ctaagtaatt aaatgacact aagtaacgaa agatatctgt aaacaacgac
      421 tatataccag aattagtcat attctctatt gtattcaccc tacacagccg cattacacag
      481 ctgctccgga cgttttacgc tgcacatcaa tgactaatac cactgtattt gatccacact
      541 tagcgcaaaa gtcacctggg tggcgttttc cgctcgcgct ccgccggtga gagcgagaaa
      601 aaaaaaaggg attgggcggt aatgtaagga aaacccacta gttaaaatca atgagataag
      661 gaactctctt tttttcccgt taatcatctt ttccttgttc gtttcgtgta tctgtaatga
      721 atcttatcgg agtaactatt actggaaagc cataccgtta aagaagcaag catatcattc
      781 gtattcaagc atctgttttt ctttttaacc gctcggctct gatttcttaa accatttccc
      841 ccatcagcat cagtttgtga tttgtaacta tatcgatcgt ggaatcgatg gaattctaag
      901 cgtcatttag ttggcaacaa aaggaatcga gcatctcata aaggagctta tattattaat
      961 atcttcctga tcgtactatt tttttttagc tttttatttt ttaggccgag gttaattcta
     1021 gaacgccaca tttcatcacc tttaagttaa gtaccaaggc catcttttac cctttttagt
     1081 ttctggggaa acagagaaca gtactataga agataaaaag atataataca atcgtagtaa
     1141 tagtacccat aataaaataa catataatat aataaaaaca taaggcaaag gagataggag
     1201 aataatcatt gatatctcca caaaacgaat ttgaagctac cccatatttt caaatctctt
     1261 ttacaacacc agacgagaaa ttaagaaact cgagatggga gacattaaaa ataaagatca
     1321 cacaacctct gtgaaccata atctaatggc aagtgcagga aattacaccg ctgagaaaga
     1381 aatcggaaag ggttcgtttg ccactgtata tagagggcat ctgacatccg acaaatctca
     1441 gcatgtagcc ataaaggaag tatcaagggc gaaattaaaa aataagaaat tactagagaa
     1501 tttggaaata gaaatcgcta tcttgaagaa aatcaagcat cctcatatcg tcggacttat
     1561 tgactgtgaa cgaacatcaa cagattttta tttgatcatg gagtactgtg ctcttgggga
     1621 cctaacattt ctgttgaaaa ggcgtaaaga attgatggag aatcatcctc tactaaggac
     1681 cgtatttgaa aaataccctc caccgagtga gaaccataac ggcctgcata gagcgtttgt
     1741 cttgagttat ttacagcagt tagcgtccgc tttgaaattt ttgaggtcta aaaacttggt
     1801 tcatagagac atcaagcctc aaaacttact attatctaca cccctaattg gatatcatga
     1861 ttcaaaaagc ttccatgaac ttggatttgt tgggatctac aacttaccca ttttaaagat
     1921 agcagatttc gggtttgcaa gatttttgcc aaacacgtca ttagcagaaa ctctttgtgg
     1981 ctcaccatta tatatggcac cagaaatttt gaattatcaa aaatataacg ctaaagcaga
     2041 tctgtggtct gtcggtacag tggtattcga aatgtgctgt ggcaccccac cgtttagagc
     2101 ttccaatcat ttggagttat tcaagaaaat taaaagagca aacgatgtca taacgtttcc
     2161 ttcatattgc aatattgaac cagagttaaa agagttgata tgtagtttat tgacatttga
     2221 tccagcccaa agaataggat ttgaggagtt ttttgctaac aaggtagtca acgaagactt
     2281 gtcttcttat gaattggaag atgatttacc tgagttagaa tccaaatcaa aaggtattgt
     2341 agaaagtaat atgttcgttt ctgagtattt atctaaacag ccaaagagtc cgaacagtaa
     2401 tcttgcaggt catcaatcaa tggcagataa tcccgcggag ctcagtgatg ccctcaagaa
     2461 cagcaatata ttaactgccc cagccgttaa aacagaccat acacaagccg tagataaaaa
     2521 ggcttcaaat aataaatacc ataatagcct agtttcagat agaagctttg aaagagaata
     2581 tgtggtggta gagaagaaat cggttgaagt taattcattg gcagacgagg ttgctcaagc
     2641 aggattcaat ccaaatccta ttaagcaccc aacttcaact caaaatcaga acgtcttatt
     2701 gaatgagcaa ttctctccaa acaatcaaca gtattttcaa aatcaaggag aaaatccgag
     2761 gttactgagg gccacatcat cttccagtgg aggtagtgat gggtctaggc gaccatcttt
     2821 ggtggataga cgcttgtcta tatcctcgct gaatccatct aatgcattat caagagccct
     2881 cggtattgca tcaacgagat tgtttggtgg tgcaaatcaa cagcagcaac agcaacaaat
     2941 cacatcttcc ccaccgtaca gtcaaacttt gttgaattcc caactttttc atgaacttac
     3001 tgagaatata atattaagaa tagatcacct acagcatcca gagacactga aattagataa
     3061 tactaatatc gttagtattt tggaatctct ggccgcaaag gcatttgttg tttactctta
     3121 tgcagaagtg aaattttctc aaattgttcc attatcaaca acattaaaag gcatggctaa
     3181 ctttgagaac aggcgcagta tggatagtaa tgctattgca gaagaacaag actcagacga
     3241 tgcagaagaa gaggatgaaa cgttgaaaaa gtacaaggaa gattgcttat ccacgaaaac
     3301 ttttggaaag ggtagaactt tatctgccac atctcagttg agtgcaactt tcaataaact
     3361 accacgttcg gaaatgatcc ttctatgtaa tgaggccatt gtcttatata tgaaggcatt
     3421 atccatttta tcaaaatcta tgcaggtaac gtccaactgg tggtatgaat ctcaagaaaa
     3481 atcatgttct ctaagagtta acgtgttggt acagtggcta agggaaaaat ttaatgaatg
     3541 tttagaaaaa gctgatttcc tgagattaaa aattaacgac ttgagattca agcatgcttc
     3601 tgaggtagct gaaaatcaaa ctttggaaga aaaaggtagt tcggaagagc cagtatattt
     3661 agaaaagcta ttatatgatc gtgcattaga aatatctaag atggccgcac atatggaatt
     3721 aaaaggagaa aatttgtaca actgtgaatt agcttatgca acttcacttt ggatgttaga
     3781 aacttctctg gacgatgatg attttacgaa tgcctatggt gactaccctt ttaaaactaa
     3841 catacatcta aaaagtaacg acgttgaaga taaagagaag tatcatagtg tgttggatga
     3901 aaatgacaga ataatcataa gaaagtatat tgatagtatt gcaaacaggt tgaaaatatt
     3961 gaggcagaag atgaaccacc aaaattaata ttttgcatat aatagcattg gtacatattg
     4021 tggtgccact ttccgtaatt aagctactaa atttttgact ctaacgttta taacattgaa
     4081 ttattaaaat gatcttgcta tactattttg catacattat tttttgtagt tttaacaaga
     4141 agacaaattc ttgactttac tcacaaaaca taaacataaa atgtaataaa gtcctcaatg
     4201 ttcaccgtat ttttcaaaaa ggatcagtga aagtgtcggt aaagctacct ttttgatact
     4261 ttctttttat gagaataagt tgtatttctt tattatgtat gatacgggcg ttcaatcaat
     4321 acgttttaag ttcgttgctg acggctaaga aattggtaag aggagtagtt ccatcaatgt
     4381 ctaaagtgat atgataaaaa ctatacgatt tgaagatcta tgcgttcact tcacgtatcc
     4441 aagtttcccc tcctaataat tcacattaat ttggcattca tatctaaatg atctgagatg
     4501 caaaagcggt ggatgatggg tacccaattc gccctatagt gagtcgtatt acgcgcgctc
     4561 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4621 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     4681 cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag cggcgcatta
     4741 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4801 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4861 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4921 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4981 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     5041 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     5101 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     5161 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     5221 gcatagggta ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat
     5281 gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc
     5341 cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt
     5401 cctcttccaa caataataat gtcagatcct gtagagacca catcatccac ggttctatac
     5461 tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa
     5521 tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct
     5581 ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc
     5641 ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc
     5701 gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct
     5761 gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg
     5821 tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc
     5881 ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa
     5941 caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc
     6001 aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca
     6061 ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt
     6121 ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct
     6181 tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc
     6241 ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa accgaaatca aaaaaaagaa
     6301 taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac tctcagtaca atctgctctg
     6361 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     6421 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     6481 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     6541 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6601 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6661 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6721 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6781 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6841 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6901 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     6961 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     7021 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     7081 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     7141 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     7201 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     7261 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     7321 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7381 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     7441 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7501 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7561 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7621 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7681 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7741 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7801 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7861 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7921 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7981 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     8041 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     8101 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     8161 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     8221 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     8281 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     8341 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     8401 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     8461 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     8521 gcccaatacg caaaccgcct ctccccgcg
//
