LOCUS       sequence-236131-        6592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing Atg17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6592)
  AUTHORS   Liu X, Mao K, Yu AYH, Omairi-Nasser A, Austin J 2nd, Glick BS, Yip 
            CK, Klionsky DJ
  TITLE     The Atg17-Atg31-Atg29 Complex Coordinates with Atg11 to Recruit the 
            Vam7 SNARE and Mediate Autophagosome-Vacuole Fusion.
  JOURNAL   Curr Biol. 2016 Jan 25;26(2):150-160. doi: 
            10.1016/j.cub.2015.11.054. Epub 2016 Jan 7.
  PUBMED    26774783
REFERENCE   2  (bases 1 to 6592)
  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.cub.2015.11.054"; journalName: "Curr Biol"; date: 
            "2016-01-25- 25"; volume: "26"; issue: "2"; pages: "150-160"
COMMENT     Sequence Label: pCu-Atg17(416) (I325D)
FEATURES             Location/Qualifiers
     source          1..6592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    66..87
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(102..107,108..125,126..132)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    140..156
                     /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     145..167
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     164..180
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     164..180
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     199..219
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        201..219
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        238..472
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     RBS             1568..1579
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1788..2035
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1804..1822)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2053..2072)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2054..2072)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2082..2099)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2082..2098)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2091..2113)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2239..2694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2326..2345)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2536..2557
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2825..3628)
                     /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(3629..3844)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3868..3887)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3987..4009
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4047..4065)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4106..4609
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4646..4750
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4751..4819,4820..5611)
                     /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(4969..4988)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5782..6370
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6271..6290
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6524..6541
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
       61 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
      121 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
      181 catgattacg ccaagcgcgc aattaaccct cactaaaggg aacaaaagct ggagctccta
      241 gttagaaaaa gacatttttg ctgtcagtca ctgtcaagag attcttttgc tggcatttct
      301 tctagaagca aaaagagcga tgcgtctttt ccgctgaacc gttccagcaa aaaagactac
      361 caacgcaata tggattgtca gaatcatata aaagagaagc aaataactcc ttgtcttgta
      421 tcaattgcat tataatatct tcttgttagt gcaatatcat atagaagtca tcgactagtg
      481 gatcccccgg gctgcaggaa ttcgatatca agcttatcga tatgaacgaa gcagatgtta
      541 caaaatttgt taataatgcc aggaaaacgc tgaccgatgc tcaactttta tgttcaagtg
      601 ctaatttaag gattgtagat attaagaaaa aattgtcatc ttggcagttg agtatttcaa
      661 aactcaattt tctaatagtt ggcttgagac agcaaggaaa gtttctttac actattttaa
      721 aggaaggcat tgggacaaag ctaatccaga aacaatggaa tcaagctgtc ttagtggtct
      781 tagttgatga gatgaagtac tggcaatatg aaatcacttc taaagtacaa aggttggatg
      841 gtatagtaaa tgaattgagc atatcagaaa aagatgatac cgatccctct aaactaggag
      901 attacatctc gagagacaac gttaacttat tgaatgacaa actgaaagaa gtgccagtaa
      961 ttgagcgtca aattgaaaac attaagcttc aatacgaaaa tatggtcaga aaagttaaca
     1021 aagaactgat tgataccaag ttgacggacg taactcaaaa attccaaagt aaatttggta
     1081 tagataacct gatggaaaca aatgttgcag agcagtttag cagggaacta acggaccttg
     1141 aaaaagattt agcagagata atgaattcat tgacacagca ctttgataag acattgctgt
     1201 tacaagataa aaaaattgat aacgatgaac gtgaggagct gtttaaggtg gtacaaggcg
     1261 acgacaaaga actatacaac attttcaaaa ctctgcatga ggtaattgat gacgtggaca
     1321 aaacaattct taacttgggt caatttttgc aggcaaaaat aaaggaaaag acagaactac
     1381 acagcgaagt ttctgaaata ataaacgatt tcaatagaaa tttggaatat ctattaatct
     1441 ttaaagatat ttccaatctg attgatagct tcaaaaattc ctgtacacaa gatgatcaaa
     1501 caactaagga actttgtgaa ttttatgata attttgaaga aagctacggt aacttagttc
     1561 tagaagcaaa gaggagaaag gatgtggcaa acagaatgaa aactatattg aaagattgtg
     1621 aaaagcagtt gcagaattta gatgctcagg accaggaaga acgtcagaat tttatagcgg
     1681 aaaatggaac ttatcttcct gagacaatct ggcccggtaa aattgacgat ttttcttccc
     1741 tgtacacttt aaattacaac gtgaagaatc cttaggtcga cctcgagtca tgtaattagt
     1801 tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt
     1861 tagacaacct gaagtctagg tccctattta tttttttata gttatgttag tattaagaac
     1921 gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat
     1981 tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcggccg
     2041 gtacccaatt cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa
     2101 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     2161 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     2221 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     2281 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     2341 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     2401 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     2461 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     2521 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     2581 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     2641 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     2701 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat
     2761 ataattaaat tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag
     2821 ttttttagtt ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac
     2881 gttcaccctc taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa
     2941 tgtcagatcc tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc
     3001 ccttgtcatc taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc
     3061 cacccatgtc tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt
     3121 caacagtacc cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta
     3181 acatcaaaag gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa
     3241 caatacctgg gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt
     3301 atacacccgc agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt
     3361 cgaagagtaa aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca
     3421 tggaaaaatc agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg
     3481 cttcaactaa ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg
     3541 tttgcttttc gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag
     3601 cacgttcctt atatgtagct ttcgacatga tttatcttcg tttcctgcag gtttttgttc
     3661 tgtgcagttg ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta
     3721 tatataccaa tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg
     3781 aatcaaaaaa atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat
     3841 tgaaaaggtg gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3901 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     3961 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4021 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4081 catgataata atggtttctt agacggatcg cttgcctgta acttacacgc gcctcgtatc
     4141 ttttaatgat ggaataattt gggaatttac tctgtgttta tttattttta tgttttgtat
     4201 ttggatttta gaaagtaaat aaagaaggta gaagagttac ggaatgaaga aaaaaaaata
     4261 aacaaaggtt taaaaaattt caacaaaaag cgtactttac atatatattt attagacaag
     4321 aaaagcagat taaatagata tacattcgat taacgataag taaaatgtaa aatcacagga
     4381 ttttcgtgtg tggtcttcta cacagacaag atgaaacaat tcggcattaa tacctgagag
     4441 caggaagagc aagataaaag gtagtatttg ttggcgatcc ccctagagtc ttttacatct
     4501 tcggaaaaca aaaactattt tttctttaat ttcttttttt actttctatt tttaatttat
     4561 atatttatat taaaaaattt aaattataat tatttttata gcacgtgatg aaaaggaccc
     4621 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4681 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4741 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4801 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4861 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4921 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4981 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5041 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5101 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5161 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5221 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5281 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5341 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     5401 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5461 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5521 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5581 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5641 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5701 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5761 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5821 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5881 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     5941 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6001 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6061 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6121 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6181 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6241 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6301 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6361 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6421 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     6481 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     6541 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cg
//
