LOCUS       sequence-240920-        9389 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 9389)
  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 9389)
  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(406)
FEATURES             Location/Qualifiers
     source          1..9389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    102..123
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(138..143,144..161,162..168)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    176..192
                     /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     181..203
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     200..216
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     200..216
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     235..255
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        237..255
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     292..308
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     292..308
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5399..5418)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5400..5418)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5428..5445)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5428..5444)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5437..5459)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5585..6040
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5672..5691)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5882..5903
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(6171..6974)
                     /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(6975..7190)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(7214..7233)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7333..7355
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7393..7411)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7479..7583
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7584..7652,7653..8444)
                     /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(7802..7821)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8615..9203
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9104..9123
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9357..9374
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
       61 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
      121 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
      181 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gcgcgcaatt
      241 aaccctcact aaagggaaca aaagctggag ctccaccgcg gtggcggccg ctctagaact
      301 agtggatccc ccgggacctg ccacaaggtt atttctacac acaataaaaa taagatacat
      361 tgccgctgct ccgcattaca tatcatatag agtgacacta agtaattaaa tgacactaag
      421 taacgaaaga tatctgtaaa caacgactat ataccagaat tagtcatatt ctctattgta
      481 ttcaccctac acagccgcat tacacagctg ctccggacgt tttacgctgc acatcaatga
      541 ctaataccac tgtatttgat ccacacttag cgcaaaagtc acctgggtgg cgttttccgc
      601 tcgcgctccg ccggtgagag cgagaaaaaa aaaagggatt gggcggtaat gtaaggaaaa
      661 cccactagtt aaaatcaatg agataaggaa ctctcttttt ttcccgttaa tcatcttttc
      721 cttgttcgtt tcgtgtatct gtaatgaatc ttatcggagt aactattact ggaaagccat
      781 accgttaaag aagcaagcat atcattcgta ttcaagcatc tgtttttctt tttaaccgct
      841 cggctctgat ttcttaaacc atttccccca tcagcatcag tttgtgattt gtaactatat
      901 cgatcgtgga atcgatggaa ttctaagcgt catttagttg gcaacaaaag gaatcgagca
      961 tctcataaag gagcttatat tattaatatc ttcctgatcg tactattttt ttttagcttt
     1021 ttatttttta ggccgaggtt aattctagaa cgccacattt catcaccttt aagttaagta
     1081 ccaaggccat cttttaccct ttttagtttc tggggaaaca gagaacagta ctatagaaga
     1141 taaaaagata taatacaatc gtagtaatag tacccataat aaaataacat ataatataat
     1201 aaaaacataa ggcaaaggag ataggagaat aatcattgat atctccacaa aacgaatttg
     1261 aagctacccc atattttcaa atctctttta caacaccaga cgagaaatta agaaactcga
     1321 gatgggagac attaaaaata aagatcacac aacctctgtg aaccataatc taatggcaag
     1381 tgcaggaaat tacaccgctg agaaagaaat cggaaagggt tcgtttgcca ctgtatatag
     1441 agggcatctg acatccgaca aatctcagca tgtagccata aaggaagtat caagggcgaa
     1501 attaaaaaat aagaaattac tagagaattt ggaaatagaa atcgctatct tgaagaaaat
     1561 caagcatcct catatcgtcg gacttattga ctgtgaacga acatcaacag atttttattt
     1621 gatcatggag tactgtgctc ttggggacct aacatttctg ttgaaaaggc gtaaagaatt
     1681 gatggagaat catcctctac taaggaccgt atttgaaaaa taccctccac cgagtgagaa
     1741 ccataacggc ctgcatagag cgtttgtctt gagttattta cagcagttag cgtccgcttt
     1801 gaaatttttg aggtctaaaa acttggttca tagagacatc aagcctcaaa acttactatt
     1861 atctacaccc ctaattggat atcatgattc aaaaagcttc catgaacttg gatttgttgg
     1921 gatctacaac ttacccattt taaagatagc agatttcggg tttgcaagat ttttgccaaa
     1981 cacgtcatta gcagaaactc tttgtggctc accattatat atggcaccag aaattttgaa
     2041 ttatcaaaaa tataacgcta aagcagatct gtggtctgtc ggtacagtgg tattcgaaat
     2101 gtgctgtggc accccaccgt ttagagcttc caatcatttg gagttattca agaaaattaa
     2161 aagagcaaac gatgtcataa cgtttccttc atattgcaat attgaaccag agttaaaaga
     2221 gttgatatgt agtttattga catttgatcc agcccaaaga ataggatttg aggagttttt
     2281 tgctaacaag gtagtcaacg aagacttgtc ttcttatgaa ttggaagatg atttacctga
     2341 gttagaatcc aaatcaaaag gtattgtaga aagtaatatg ttcgtttctg agtatttatc
     2401 taaacagcca aagagtccga acagtaatct tgcaggtcat caatcaatgg cagataatcc
     2461 cgcggagctc agtgatgccc tcaagaacag caatatatta actgccccag ccgttaaaac
     2521 agaccataca caagccgtag ataaaaaggc ttcaaataat aaataccata atagcctagt
     2581 ttcagataga agctttgaaa gagaatatgt ggtggtagag aagaaatcgg ttgaagttaa
     2641 ttcattggca gacgaggttg ctcaagcagg attcaatcca aatcctatta agcacccaac
     2701 ttcaactcaa aatcagaacg tcttattgaa tgagcaattc tctccaaaca atcaacagta
     2761 ttttcaaaat caaggagaaa atccgaggtt actgagggcc acatcatctt ccagtggagg
     2821 tagtgatggg tctaggcgac catctttggt ggatagacgc ttgtctatat cctcgctgaa
     2881 tccatctaat gcattatcaa gagccctcgg tattgcatca acgagattgt ttggtggtgc
     2941 aaatcaacag cagcaacagc aacaaatcac atcttcccca ccgtacagtc aaactttgtt
     3001 gaattcccaa ctttttcatg aacttactga gaatataata ttaagaatag atcacctaca
     3061 gcatccagag acactgaaat tagataatac taatatcgtt agtattttgg aatctctggc
     3121 cgcaaaggca tttgttgttt actcttatgc agaagtgaaa ttttctcaaa ttgttccatt
     3181 atcaacaaca ttaaaaggca tggctaactt tgagaacagg cgcagtatgg atagtaatgc
     3241 tattgcagaa gaacaagact cagacgatgc agaagaagag gatgaaacgt tgaaaaagta
     3301 caaggaagat tgcttatcca cgaaaacttt tggaaagggt agaactttat ctgccacatc
     3361 tcagttgagt gcaactttca ataaactacc acgttcggaa atgatccttc tatgtaatga
     3421 ggccattgtc ttatatatga aggcattatc cattttatca aaatctatgc aggtaacgtc
     3481 caactggtgg tatgaatctc aagaaaaatc atgttctcta agagttaacg tgttggtaca
     3541 gtggctaagg gaaaaattta atgaatgttt agaaaaagct gatttcctga gattaaaaat
     3601 taacgacttg agattcaagc atgcttctga ggtagctgaa aatcaaactt tggaagaaaa
     3661 aggtagttcg gaagagccag tatatttaga aaagctatta tatgatcgtg cattagaaat
     3721 atctaagatg gccgcacata tggaattaaa aggagaaaat ttgtacaact gtgaattagc
     3781 ttatgcaact tcactttgga tgttagaaac ttctctggac gatgatgatt ttacgaatgc
     3841 ctatggtgac taccctttta aaactaacat acatctaaaa agtaacgacg ttgaagataa
     3901 agagaagtat catagtgtgt tggatgaaaa tgacagaata atcataagaa agtatattga
     3961 tagtattgca aacaggttga aaatattgag gcagaagatg aaccaccaaa attaagtgat
     4021 gcataatatg gttttcttat taagtacaaa tgacctgcta tattttcaag tatcttcatg
     4081 attatgttat gctactctcg aaatgcttta cgagttcgtc attaaagtat tcaaatttta
     4141 ttttcccttt gtgttttgtt ttgtttttat ttctacgtga tttttttctt gaaatccagc
     4201 tcttttatac ctggatgggt tttattttga tgtttaaatg ttaattaagg aaattattga
     4261 attccttcgg tttcggtgtg tttttttctt tttaagtttt tttttgttgc gtttagtact
     4321 tgcctttggc tatacatatc cttttaatct aatttttatt tcttcaaaat ttgattcttt
     4381 aaccttttgt tctatatata tatataaatg tatgtatgta tgatttgatt gaggaggctt
     4441 taacaagtca ttaatatctt tattattttt acttgttgag ttcattaaaa ttcattttat
     4501 aatttgttgt tacaaacagt attttacaag tcataataaa cgttcaaaaa caaacacaat
     4561 attaaaagaa ttggtattag gatccgacat gacaattaac ttagatcctt ctatcaagtt
     4621 tctaatcata tattacatct attaaaataa tttacatata tcatgctaaa gcttataaag
     4681 agacattctt cttctcgtgc gctgggcgtc gctagcctca aatacaagtg atgcgtctgc
     4741 ttctattcta aggtcatcct tgctgttaga tttcccttct accaggtggg cttttatacg
     4801 ttgaggtaca ttaccgacag gcgtaagagt agagctcaaa cttgcttccg tactctcaaa
     4861 gggatattct cctacaggag acatagttgt atattgtgga aactcacaag aattcactct
     4921 taatgaactt ccactctcac ctatcaagca ttgttgatca ccgattgctg gggcatcctg
     4981 aatactgatc tgagatgcaa aagcggtgga tgatgggtac cgtactggtc atacaaagct
     5041 cttaaacgat tgtctttaag tatttcgtat gcctggcata tctcttgaaa ctttcgcttg
     5101 gattcttctg tatgattgtt tttatcagga tgatatttta atgcagcgtt cctgtaagct
     5161 ttcttaatct cagatgtgga tgcaaaagcg gtggatgatg ggtaccgtac tggtcataca
     5221 aagctcttaa acgattgtct ttaagtattt cgtatgcctg gcatatctct tgaaactttc
     5281 gcttggattc ttctgtatga ttgtttttat caggatgata ttttaatgca gcgttcctgt
     5341 aagctttctt aatctcagat gtggatgcaa aagcggtgga tgatgggtac ccaattcgcc
     5401 ctatagtgag tcgtattacg cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga
     5461 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     5521 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     5581 atggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     5641 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     5701 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     5761 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     5821 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     5881 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     5941 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     6001 tttaacgcga attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc
     6061 ttacgcatct gtgcggtatt tcacaccgca tagggtaata actgatataa ttaaattgaa
     6121 gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt ttagttttgc
     6181 tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc accctctacc
     6241 ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc agatcctgta
     6301 gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt gtcatctaaa
     6361 cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc catgtctctt
     6421 tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac agtaccctta
     6481 gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat caaaaggcct
     6541 ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat acctgggccc
     6601 accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac acccgcagag
     6661 tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa
     6721 ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga aaaatcagtc
     6781 aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc aactaactcc
     6841 agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg cttttcgtgc
     6901 atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg ttccttatat
     6961 gtagctttcg acatgattta tcttcgtttc ctgcaggttt ttgttctgtg cagttgggtt
     7021 aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata taccaatcta
     7081 agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt
     7141 caaggaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa aaggtggtat
     7201 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc
     7261 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     7321 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     7381 cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     7441 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     7501 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     7561 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     7621 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     7681 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     7741 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     7801 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     7861 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     7921 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     7981 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     8041 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     8101 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     8161 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     8221 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     8281 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     8341 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     8401 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     8461 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     8521 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     8581 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     8641 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     8701 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     8761 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     8821 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     8881 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     8941 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     9001 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     9061 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     9121 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     9181 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     9241 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     9301 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     9361 agtcagtgag cgaggaagcg gaagagcgc
//
