LOCUS       sequence-240333-        8659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing ATG1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8659)
  AUTHORS   Liu X, Yao Z, Jin M, Namkoong S, Yin Z, Lee JH, Klionsky DJ
  TITLE     Dhh1 promotes autophagy-related protein translation during nitrogen 
            starvation.
  JOURNAL   PLoS Biol. 2019 Apr 11;17(4):e3000219. doi: 
            10.1371/journal.pbio.3000219. eCollection 2019 Apr.
  PUBMED    30973873
REFERENCE   2  (bases 1 to 8659)
  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"; journalName: "PLoS 
            Biol."; date: "2019-04-11"; volume: "17(4):e3000219. doi"; pages: " 
            10.1371/journal.pbio.3000219. eCollection 2019 Apr"
COMMENT     Sequence Label: pRS-ATG7p-ATG1(406)
FEATURES             Location/Qualifiers
     source          1..8659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(131..136,137..154,155..161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    169..185
                     /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     174..196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     193..209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     193..209
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     228..248
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        230..248
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     285..301
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     285..301
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4662..4681)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4663..4681)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4691..4708)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4691..4707)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4700..4722)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4848..5303
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4935..4954)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5145..5166
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5434..6237)
                     /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(6238..6453)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6477..6496)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6596..6618
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6656..6674)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6742..6846
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6847..6915,6916..7707)
                     /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(7065..7084)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7878..8466
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8367..8386
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8620..8637
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      181 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      241 actaaaggga acaaaagctg gagctccacc gcggtggcgg ccgctctaga actagtggat
      301 cccccgggct gcaggaattc gatatcaagc ttatcgatgc atccttagat tacaattcaa
      361 gagctcaaaa gcaagaaatg gaacgtgcag aaaaggacta cgaattattc ttacaagaat
      421 tagaagagga tgctgaatta agacaaagtg ttaacctata caaaaaccgc gaagcaaatg
      481 tacctcctga agagcatgaa atggatgaag acgaggatga agacgctcca caaatcaaca
      541 ttgatgaatt attggacgag ttagatgaaa tgacattaga agatggagtc gagaacacac
      601 ccgttgaatc tcagcagtaa aaaaacacca aagaatggag agtagtatat actcttaaca
      661 aacactagca tttcagttta tctatataca tatatatgga tgtaaaatgt actttatgga
      721 agaacaagcc accacatgtt gaaaactaga taggcaagca agatttttca tttgtagagc
      781 tcttacgcaa gatttttaaa cttccgcttt tttaatccat gagattcctt tggacaccct
      841 ttccggcacg gcaaaacaaa aaaattaagg gaactcatta ttttacgatg ctacttagat
      901 aactaaagtt cattatattt caacaaatat aagataatca agaataaact cgagatggga
      961 gacattaaaa ataaagatca cacaacctct gtgaaccata atctaatggc aagtgcagga
     1021 aattacaccg ctgagaaaga aatcggaaag ggttcgtttg ccactgtata tagagggcat
     1081 ctgacatccg acaaatctca gcatgtagcc ataaaggaag tatcaagggc gaaattaaaa
     1141 aataagaaat tactagagaa tttggaaata gaaatcgcta tcttgaagaa aatcaagcat
     1201 cctcatatcg tcggacttat tgactgtgaa cgaacatcaa cagattttta tttgatcatg
     1261 gagtactgtg ctcttgggga cctaacattt ctgttgaaaa ggcgtaaaga attgatggag
     1321 aatcatcctc tactaaggac cgtatttgaa aaataccctc caccgagtga gaaccataac
     1381 ggcctgcata gagcgtttgt cttgagttat ttacagcagt tagcgtccgc tttgaaattt
     1441 ttgaggtcta aaaacttggt tcatagagac atcaagcctc aaaacttact attatctaca
     1501 cccctaattg gatatcatga ttcaaaaagc ttccatgaac ttggatttgt tgggatctac
     1561 aacttaccca ttttaaagat agcagatttc gggtttgcaa gatttttgcc aaacacgtca
     1621 ttagcagaaa ctctttgtgg ctcaccatta tatatggcac cagaaatttt gaattatcaa
     1681 aaatataacg ctaaagcaga tctgtggtct gtcggtacag tggtattcga aatgtgctgt
     1741 ggcaccccac cgtttagagc ttccaatcat ttggagttat tcaagaaaat taaaagagca
     1801 aacgatgtca taacgtttcc ttcatattgc aatattgaac cagagttaaa agagttgata
     1861 tgtagtttat tgacatttga tccagcccaa agaataggat ttgaggagtt ttttgctaac
     1921 aaggtagtca acgaagactt gtcttcttat gaattggaag atgatttacc tgagttagaa
     1981 tccaaatcaa aaggtattgt agaaagtaat atgttcgttt ctgagtattt atctaaacag
     2041 ccaaagagtc cgaacagtaa tcttgcaggt catcaatcaa tggcagataa tcccgcggag
     2101 ctcagtgatg ccctcaagaa cagcaatata ttaactgccc cagccgttaa aacagaccat
     2161 acacaagccg tagataaaaa ggcttcaaat aataaatacc ataatagcct agtttcagat
     2221 agaagctttg aaagagaata tgtggtggta gagaagaaat cggttgaagt taattcattg
     2281 gcagacgagg ttgctcaagc aggattcaat ccaaatccta ttaagcaccc aacttcaact
     2341 caaaatcaga acgtcttatt gaatgagcaa ttctctccaa acaatcaaca gtattttcaa
     2401 aatcaaggag aaaatccgag gttactgagg gccacatcat cttccagtgg aggtagtgat
     2461 gggtctaggc gaccatcttt ggtggataga cgcttgtcta tatcctcgct gaatccatct
     2521 aatgcattat caagagccct cggtattgca tcaacgagat tgtttggtgg tgcaaatcaa
     2581 cagcagcaac agcaacaaat cacatcttcc ccaccgtaca gtcaaacttt gttgaattcc
     2641 caactttttc atgaacttac tgagaatata atattaagaa tagatcacct acagcatcca
     2701 gagacactga aattagataa tactaatatc gttagtattt tggaatctct ggccgcaaag
     2761 gcatttgttg tttactctta tgcagaagtg aaattttctc aaattgttcc attatcaaca
     2821 acattaaaag gcatggctaa ctttgagaac aggcgcagta tggatagtaa tgctattgca
     2881 gaagaacaag actcagacga tgcagaagaa gaggatgaaa cgttgaaaaa gtacaaggaa
     2941 gattgcttat ccacgaaaac ttttggaaag ggtagaactt tatctgccac atctcagttg
     3001 agtgcaactt tcaataaact accacgttcg gaaatgatcc ttctatgtaa tgaggccatt
     3061 gtcttatata tgaaggcatt atccatttta tcaaaatcta tgcaggtaac gtccaactgg
     3121 tggtatgaat ctcaagaaaa atcatgttct ctaagagtta acgtgttggt acagtggcta
     3181 agggaaaaat ttaatgaatg tttagaaaaa gctgatttcc tgagattaaa aattaacgac
     3241 ttgagattca agcatgcttc tgaggtagct gaaaatcaaa ctttggaaga aaaaggtagt
     3301 tcggaagagc cagtatattt agaaaagcta ttatatgatc gtgcattaga aatatctaag
     3361 atggccgcac atatggaatt aaaaggagaa aatttgtaca actgtgaatt agcttatgca
     3421 acttcacttt ggatgttaga aacttctctg gacgatgatg attttacgaa tgcctatggt
     3481 gactaccctt ttaaaactaa catacatcta aaaagtaacg acgttgaaga taaagagaag
     3541 tatcatagtg tgttggatga aaatgacaga ataatcataa gaaagtatat tgatagtatt
     3601 gcaaacaggt tgaaaatatt gaggcagaag atgaaccacc aaaattaagt gatgcataat
     3661 atggttttct tattaagtac aaatgacctg ctatattttc aagtatcttc atgattatgt
     3721 tatgctactc tcgaaatgct ttacgagttc gtcattaaag tattcaaatt ttattttccc
     3781 tttgtgtttt gttttgtttt tatttctacg tgattttttt cttgaaatcc agctctttta
     3841 tacctggatg ggttttattt tgatgtttaa atgttaatta aggaaattat tgaattcctt
     3901 cggtttcggt gtgttttttt ctttttaagt ttttttttgt tgcgtttagt acttgccttt
     3961 ggctatacat atccttttaa tctaattttt atttcttcaa aatttgattc tttaaccttt
     4021 tgttctatat atatatataa atgtatgtat gtatgatttg attgaggagg ctttaacaag
     4081 tcattaatat ctttattatt tttacttgtt gagttcatta aaattcattt tataatttgt
     4141 tgttacaaac agtattttac aagtcataat aaacgttcaa aaacaaacac aatattaaaa
     4201 gaattggtat taggatccga catgacaatt aacttagatc cttctatcaa gtttctaatc
     4261 atatattaca tctattaaaa taatttacat atatcatgct aaagcttata aagagacatt
     4321 cttcttctcg tgcgctgggc gtcgctagcc tcaaatacaa gtgatgcgtc tgcttctatt
     4381 ctaaggtcat ccttgctgtt agatttccct tctaccaggt gggcttttat acgttgaggt
     4441 acattaccga caggcgtaag agtagagctc aaacttgctt ccgtactctc aaagggatat
     4501 tctcctacag gagacatagt tgtatattgt ggaaactcac aagaattcac tcttaatgaa
     4561 cttccactct cacctatcaa gcattgttga tcaccgattg ctggggcatc ctgaatactg
     4621 atctgagatg caaaagcggt ggatgatggg tacccaattc gccctatagt gagtcgtatt
     4681 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     4741 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     4801 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
     4861 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     4921 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     4981 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5041 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5101 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5161 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     5221 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5281 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     5341 atttcacacc gcatagggta ataactgata taattaaatt gaagctctaa tttgtgagtt
     5401 tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa
     5461 atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt
     5521 tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca catcatccac
     5581 ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat
     5641 aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat
     5701 aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga
     5761 tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac
     5821 ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt
     5881 cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt
     5941 attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa
     6001 tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt
     6061 ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt
     6121 acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt
     6181 ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat
     6241 ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata ctgggcaatt
     6301 tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct
     6361 tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa accgaaatca
     6421 aaaaaaagaa taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac tctcagtaca
     6481 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     6541 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     6601 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     6661 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     6721 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     6781 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6841 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6901 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6961 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7021 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7081 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7141 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7201 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7261 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7321 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7381 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7441 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7501 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7561 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     7621 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     7681 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     7741 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7801 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7861 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     7921 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     7981 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8041 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8101 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8161 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8221 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8281 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8341 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     8401 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     8461 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     8521 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     8581 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     8641 gcggaagagc gcccaatac
//
