LOCUS       sequence-236134-        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) (F317D)
FEATURES             Location/Qualifiers
     source          1..6592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    73..94
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(109..114,115..132,133..139)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    147..163
                     /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     152..174
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     171..187
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     171..187
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     206..226
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        208..226
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        245..479
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     RBS             1575..1586
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1795..2042
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1811..1829)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2060..2079)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2061..2079)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2089..2106)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2089..2105)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2098..2120)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2246..2701
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2333..2352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2543..2564
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2832..3635)
                     /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(3636..3851)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3875..3894)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3994..4016
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4054..4072)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4113..4616
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4653..4757
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4758..4826,4827..5618)
                     /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(4976..4995)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5789..6377
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6278..6297
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6531..6548
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
       61 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
      121 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
      181 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga
      241 gctcctagtt agaaaaagac atttttgctg tcagtcactg tcaagagatt cttttgctgg
      301 catttcttct agaagcaaaa agagcgatgc gtcttttccg ctgaaccgtt ccagcaaaaa
      361 agactaccaa cgcaatatgg attgtcagaa tcatataaaa gagaagcaaa taactccttg
      421 tcttgtatca attgcattat aatatcttct tgttagtgca atatcatata gaagtcatcg
      481 actagtggat cccccgggct gcaggaattc gatatcaagc ttatcgatat gaacgaagca
      541 gatgttacaa aatttgttaa taatgccagg aaaacgctga ccgatgctca acttttatgt
      601 tcaagtgcta atttaaggat tgtagatatt aagaaaaaat tgtcatcttg gcagttgagt
      661 atttcaaaac tcaattttct aatagttggc ttgagacagc aaggaaagtt tctttacact
      721 attttaaagg aaggcattgg gacaaagcta atccagaaac aatggaatca agctgtctta
      781 gtggtcttag ttgatgagat gaagtactgg caatatgaaa tcacttctaa agtacaaagg
      841 ttggatggta tagtaaatga attgagcata tcagaaaaag atgataccga tccctctaaa
      901 ctaggagatt acatctcgag agacaacgtt aacttattga atgacaaact gaaagaagtg
      961 ccagtaattg agcgtcaaat tgaaaacatt aagcttcaat acgaaaatat ggtcagaaaa
     1021 gttaacaaag aactgattga taccaagttg acggacgtaa ctcaaaaatt ccaaagtaaa
     1081 tttggtatag ataacctgat ggaaacaaat gttgcagagc agtttagcag ggaactaacg
     1141 gaccttgaaa aagatttagc agagataatg aattcattga cacagcactt tgataagaca
     1201 ttgctgttac aagataaaaa aattgataac gatgaacgtg aggagctgtt taaggtggta
     1261 caaggcgacg acaaagaact atacaacatt ttcaaaactc tgcatgaggt aattgatgac
     1321 gtggacaaaa caattcttaa cttgggtcaa tttttgcagg caaaaataaa ggaaaagaca
     1381 gaactacaca gcgaagtttc tgaaataata aacgatttca atagaaattt ggaatatcta
     1441 ttaatcttta aagatatttc caatctgatt gatagcgaca aaaattcctg tacacaagat
     1501 attcaaacaa ctaaggaact ttgtgaattt tatgataatt ttgaagaaag ctacggtaac
     1561 ttagttctag aagcaaagag gagaaaggat gtggcaaaca gaatgaaaac tatattgaaa
     1621 gattgtgaaa agcagttgca gaatttagat gctcaggacc aggaagaacg tcagaatttt
     1681 atagcggaaa atggaactta tcttcctgag acaatctggc ccggtaaaat tgacgatttt
     1741 tcttccctgt acactttaaa ttacaacgtg aagaatcctt aggtcgacct cgagtcatgt
     1801 aattagttat gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg
     1861 aaggagttag acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat
     1921 taagaacgtt atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat
     1981 gtaacattat actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt
     2041 gcggccggta cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt
     2101 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     2161 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     2221 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2281 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2341 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2401 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2461 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2521 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2581 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2641 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     2701 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtaat
     2761 aactgatata attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat
     2821 aatacagttt tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt
     2881 ctgtaacgtt caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca
     2941 ataataatgt cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat
     3001 gcgtctccct tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca
     3061 tctcttccac ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc
     3121 gcaatgtcaa cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat
     3181 tctgctaaca tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa
     3241 ccgctaacaa tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct
     3301 attctgtata cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt
     3361 ctgtcttcga agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg
     3421 ccctccatgg aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga
     3481 cctaatgctt caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac
     3541 aagtttgttt gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga
     3601 gtagcagcac gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt
     3661 tttgttctgt gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat
     3721 atgcgtatat ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag
     3781 attaccgaat caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat
     3841 gatgaattga aaaggtggta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3901 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3961 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4021 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     4081 ttaatgtcat gataataatg gtttcttaga cggatcgctt gcctgtaact tacacgcgcc
     4141 tcgtatcttt taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt
     4201 tttgtatttg gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa
     4261 aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt
     4321 agacaagaaa agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat
     4381 cacaggattt tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac
     4441 ctgagagcag gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt
     4501 tacatcttcg gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt
     4561 aatttatata tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa
     4621 aggacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4681 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4741 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4801 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4861 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4921 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4981 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5041 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5101 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5161 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5221 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5281 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5341 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5401 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5461 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5521 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5581 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5641 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5701 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5761 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5821 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5881 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5941 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6001 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6061 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6121 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6181 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6241 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6301 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6361 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6421 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6481 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6541 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cg
//
