LOCUS       sequence-240335-        9019 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing ATG1 with ATG7 3'UTR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9019)
  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 9019)
  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-ATG1-ATG7 3'UTR(406)
FEATURES             Location/Qualifiers
     source          1..9019
                     /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(5022..5041)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5023..5041)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5051..5068)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5051..5067)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5060..5082)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5208..5663
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5295..5314)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5505..5526
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5794..6597)
                     /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(6598..6813)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6837..6856)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6956..6978
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7016..7034)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7102..7206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7207..7275,7276..8067)
                     /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(7425..7444)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8238..8826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8727..8746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8980..8997
                     /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 cccccgggac ctgccacaag gttatttcta cacacaataa aaataagata cattgccgct
      361 gctccgcatt acatatcata tagagtgaca ctaagtaatt aaatgacact aagtaacgaa
      421 agatatctgt aaacaacgac tatataccag aattagtcat attctctatt gtattcaccc
      481 tacacagccg cattacacag ctgctccgga cgttttacgc tgcacatcaa tgactaatac
      541 cactgtattt gatccacact tagcgcaaaa gtcacctggg tggcgttttc cgctcgcgct
      601 ccgccggtga gagcgagaaa aaaaaaaggg attgggcggt aatgtaagga aaacccacta
      661 gttaaaatca atgagataag gaactctctt tttttcccgt taatcatctt ttccttgttc
      721 gtttcgtgta tctgtaatga atcttatcgg agtaactatt actggaaagc cataccgtta
      781 aagaagcaag catatcattc gtattcaagc atctgttttt ctttttaacc gctcggctct
      841 gatttcttaa accatttccc ccatcagcat cagtttgtga tttgtaacta tatcgatcgt
      901 ggaatcgatg gaattctaag cgtcatttag ttggcaacaa aaggaatcga gcatctcata
      961 aaggagctta tattattaat atcttcctga tcgtactatt tttttttagc tttttatttt
     1021 ttaggccgag gttaattcta gaacgccaca tttcatcacc tttaagttaa gtaccaaggc
     1081 catcttttac cctttttagt ttctggggaa acagagaaca gtactataga agataaaaag
     1141 atataataca atcgtagtaa tagtacccat aataaaataa catataatat aataaaaaca
     1201 taaggcaaag gagataggag aataatcatt gatatctcca caaaacgaat ttgaagctac
     1261 cccatatttt caaatctctt ttacaacacc agacgagaaa ttaagaaact cgagatggga
     1321 gacattaaaa ataaagatca cacaacctct gtgaaccata atctaatggc aagtgcagga
     1381 aattacaccg ctgagaaaga aatcggaaag ggttcgtttg ccactgtata tagagggcat
     1441 ctgacatccg acaaatctca gcatgtagcc ataaaggaag tatcaagggc gaaattaaaa
     1501 aataagaaat tactagagaa tttggaaata gaaatcgcta tcttgaagaa aatcaagcat
     1561 cctcatatcg tcggacttat tgactgtgaa cgaacatcaa cagattttta tttgatcatg
     1621 gagtactgtg ctcttgggga cctaacattt ctgttgaaaa ggcgtaaaga attgatggag
     1681 aatcatcctc tactaaggac cgtatttgaa aaataccctc caccgagtga gaaccataac
     1741 ggcctgcata gagcgtttgt cttgagttat ttacagcagt tagcgtccgc tttgaaattt
     1801 ttgaggtcta aaaacttggt tcatagagac atcaagcctc aaaacttact attatctaca
     1861 cccctaattg gatatcatga ttcaaaaagc ttccatgaac ttggatttgt tgggatctac
     1921 aacttaccca ttttaaagat agcagatttc gggtttgcaa gatttttgcc aaacacgtca
     1981 ttagcagaaa ctctttgtgg ctcaccatta tatatggcac cagaaatttt gaattatcaa
     2041 aaatataacg ctaaagcaga tctgtggtct gtcggtacag tggtattcga aatgtgctgt
     2101 ggcaccccac cgtttagagc ttccaatcat ttggagttat tcaagaaaat taaaagagca
     2161 aacgatgtca taacgtttcc ttcatattgc aatattgaac cagagttaaa agagttgata
     2221 tgtagtttat tgacatttga tccagcccaa agaataggat ttgaggagtt ttttgctaac
     2281 aaggtagtca acgaagactt gtcttcttat gaattggaag atgatttacc tgagttagaa
     2341 tccaaatcaa aaggtattgt agaaagtaat atgttcgttt ctgagtattt atctaaacag
     2401 ccaaagagtc cgaacagtaa tcttgcaggt catcaatcaa tggcagataa tcccgcggag
     2461 ctcagtgatg ccctcaagaa cagcaatata ttaactgccc cagccgttaa aacagaccat
     2521 acacaagccg tagataaaaa ggcttcaaat aataaatacc ataatagcct agtttcagat
     2581 agaagctttg aaagagaata tgtggtggta gagaagaaat cggttgaagt taattcattg
     2641 gcagacgagg ttgctcaagc aggattcaat ccaaatccta ttaagcaccc aacttcaact
     2701 caaaatcaga acgtcttatt gaatgagcaa ttctctccaa acaatcaaca gtattttcaa
     2761 aatcaaggag aaaatccgag gttactgagg gccacatcat cttccagtgg aggtagtgat
     2821 gggtctaggc gaccatcttt ggtggataga cgcttgtcta tatcctcgct gaatccatct
     2881 aatgcattat caagagccct cggtattgca tcaacgagat tgtttggtgg tgcaaatcaa
     2941 cagcagcaac agcaacaaat cacatcttcc ccaccgtaca gtcaaacttt gttgaattcc
     3001 caactttttc atgaacttac tgagaatata atattaagaa tagatcacct acagcatcca
     3061 gagacactga aattagataa tactaatatc gttagtattt tggaatctct ggccgcaaag
     3121 gcatttgttg tttactctta tgcagaagtg aaattttctc aaattgttcc attatcaaca
     3181 acattaaaag gcatggctaa ctttgagaac aggcgcagta tggatagtaa tgctattgca
     3241 gaagaacaag actcagacga tgcagaagaa gaggatgaaa cgttgaaaaa gtacaaggaa
     3301 gattgcttat ccacgaaaac ttttggaaag ggtagaactt tatctgccac atctcagttg
     3361 agtgcaactt tcaataaact accacgttcg gaaatgatcc ttctatgtaa tgaggccatt
     3421 gtcttatata tgaaggcatt atccatttta tcaaaatcta tgcaggtaac gtccaactgg
     3481 tggtatgaat ctcaagaaaa atcatgttct ctaagagtta acgtgttggt acagtggcta
     3541 agggaaaaat ttaatgaatg tttagaaaaa gctgatttcc tgagattaaa aattaacgac
     3601 ttgagattca agcatgcttc tgaggtagct gaaaatcaaa ctttggaaga aaaaggtagt
     3661 tcggaagagc cagtatattt agaaaagcta ttatatgatc gtgcattaga aatatctaag
     3721 atggccgcac atatggaatt aaaaggagaa aatttgtaca actgtgaatt agcttatgca
     3781 acttcacttt ggatgttaga aacttctctg gacgatgatg attttacgaa tgcctatggt
     3841 gactaccctt ttaaaactaa catacatcta aaaagtaacg acgttgaaga taaagagaag
     3901 tatcatagtg tgttggatga aaatgacaga ataatcataa gaaagtatat tgatagtatt
     3961 gcaaacaggt tgaaaatatt gaggcagaag atgaaccacc aaaattaata ttttgcatat
     4021 aatagcattg gtacatattg tggtgccact ttccgtaatt aagctactaa atttttgact
     4081 ctaacgttta taacattgaa ttattaaaat gatcttgcta tactattttg catacattat
     4141 tttttgtagt tttaacaaga agacaaattc ttgactttac tcacaaaaca taaacataaa
     4201 atgtaataaa gtcctcaatg ttcaccgtat ttttcaaaaa ggatcagtga aagtgtcggt
     4261 aaagctacct ttttgatact ttctttttat gagaataagt tgtatttctt tattatgtat
     4321 gatacgggcg ttcaatcaat acgttttaag ttcgttgctg acggctaaga aattggtaag
     4381 aggagtagtt ccatcaatgt ctaaagtgat atgataaaaa ctatacgatt tgaagatcta
     4441 tgcgttcact tcacgtatcc aagtttcccc tcctaataat tcacattaat ttggcattca
     4501 tatctaaagc gttgctatta cccttcccct ggtagtcgcg gaacaacaaa aaacgccaaa
     4561 ataagtaatg aaaatgtaaa caaaaaggaa ctaagttgaa aagtcatcga aagcgatacc
     4621 acctggttct ttcttcatta tagactaatt gttgcgtgcc aagaagttgt ttgtgtatcc
     4681 ataatattta ttatcaaact gatctaagat tgctttgtac acatcttgcg tgaaaaaatg
     4741 gaaatcaaag aggtagatga cagggcagag ctactgagat ataccaacaa tattccgtta
     4801 cttggaaagc tagtcaacca tcaaccatta tggagtacaa atcctaagtt aaagtccttt
     4861 tcgctagaaa aaatatcagc ccctgatcaa aggcgtgtac aagaggcatt agttgtaaaa
     4921 gacctattaa atgtattgat tggacttgaa ggaacataca tcagatattt taatgattat
     4981 gaacctagtg acccagagac accgataggg tacccaattc gccctatagt gagtcgtatt
     5041 acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     5101 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     5161 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg cgccctgtag
     5221 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5281 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5341 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5401 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     5461 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5521 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     5581 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5641 caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt
     5701 atttcacacc gcatagggta ataactgata taattaaatt gaagctctaa tttgtgagtt
     5761 tagtatacat gcatttactt ataatacagt tttttagttt tgctggccgc atcttctcaa
     5821 atatgcttcc cagcctgctt ttctgtaacg ttcaccctct accttagcat cccttccctt
     5881 tgcaaatagt cctcttccaa caataataat gtcagatcct gtagagacca catcatccac
     5941 ggttctatac tgttgaccca atgcgtctcc cttgtcatct aaacccacac cgggtgtcat
     6001 aatcaaccaa tcgtaacctt catctcttcc acccatgtct ctttgagcaa taaagccgat
     6061 aacaaaatct ttgtcgctct tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga
     6121 tagggagccc ttgcatgaca attctgctaa catcaaaagg cctctaggtt cctttgttac
     6181 ttcttctgcc gcctgcttca aaccgctaac aatacctggg cccaccacac cgtgtgcatt
     6241 cgtaatgtct gcccattctg ctattctgta tacacccgca gagtactgca atttgactgt
     6301 attaccaatg tcagcaaatt ttctgtcttc gaagagtaaa aaattgtact tggcggataa
     6361 tgcctttagc ggcttaactg tgccctccat ggaaaaatca gtcaagatat ccacatgtgt
     6421 ttttagtaaa caaattttgg gacctaatgc ttcaactaac tccagtaatt ccttggtggt
     6481 acgaacatcc aatgaagcac acaagtttgt ttgcttttcg tgcatgatat taaatagctt
     6541 ggcagcaaca ggactaggat gagtagcagc acgttcctta tatgtagctt tcgacatgat
     6601 ttatcttcgt ttcctgcagg tttttgttct gtgcagttgg gttaagaata ccgggcaatt
     6661 tcatgtttct tcaacactac atatgcgtat atataccaat ctaagtctgt gctccttcct
     6721 tcgttcttcc ttctgttcgg agattaccga atcaaaaaaa tttcaaggaa accgaaatca
     6781 aaaaaaagaa taaaaaaaaa atgatgaatt gaaaaggtgg tatggtgcac tctcagtaca
     6841 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     6901 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     6961 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     7021 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     7081 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     7141 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     7201 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     7261 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7321 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7381 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7441 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7501 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7561 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7621 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7681 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7741 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7801 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7861 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7921 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     7981 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     8041 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     8101 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     8161 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8221 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8281 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8341 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8401 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     8461 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8521 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8581 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8641 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8701 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     8761 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     8821 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     8881 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     8941 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     9001 gcggaagagc gcccaatac
//
