LOCUS       sequence-236136-        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) (L105D)
FEATURES             Location/Qualifiers
     source          1..6592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    123..144
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(159..164,165..182,183..189)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    197..213
                     /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     202..224
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     221..237
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     221..237
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     256..276
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        258..276
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        295..529
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     RBS             1625..1636
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1845..2092
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1861..1879)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2110..2129)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2111..2129)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2139..2156)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2139..2155)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2148..2170)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2296..2751
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2383..2402)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2593..2614
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2882..3685)
                     /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(3686..3901)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3925..3944)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4044..4066
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4104..4122)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4163..4666
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4703..4807
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4808..4876,4877..5668)
                     /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(5026..5045)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5839..6427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6328..6347
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(6581..6592,1..6)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
       61 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      121 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
      181 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
      241 gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctgga gctcctagtt
      301 agaaaaagac atttttgctg tcagtcactg tcaagagatt cttttgctgg catttcttct
      361 agaagcaaaa agagcgatgc gtcttttccg ctgaaccgtt ccagcaaaaa agactaccaa
      421 cgcaatatgg attgtcagaa tcatataaaa gagaagcaaa taactccttg tcttgtatca
      481 attgcattat aatatcttct tgttagtgca atatcatata gaagtcatcg actagtggat
      541 cccccgggct gcaggaattc gatatcaagc ttatcgatat gaacgaagca gatgttacaa
      601 aatttgttaa taatgccagg aaaacgctga ccgatgctca acttttatgt tcaagtgcta
      661 atttaaggat tgtagatatt aagaaaaaat tgtcatcttg gcagttgagt atttcaaaac
      721 tcaattttct aatagttggc ttgagacagc aaggaaagtt tctttacact attttaaagg
      781 aaggcattgg gacaaagcta atccagaaac aatggaatca agctgtctta gtggtcttag
      841 ttgatgagat gaagtactgg caatatgaaa tcacttctaa agtacaaagg gacgatggta
      901 tagtaaatga attgagcata tcagaaaaag atgataccga tccctctaaa ctaggagatt
      961 acatctcgag agacaacgtt aacttattga atgacaaact gaaagaagtg ccagtaattg
     1021 agcgtcaaat tgaaaacatt aagcttcaat acgaaaatat ggtcagaaaa gttaacaaag
     1081 aactgattga taccaagttg acggacgtaa ctcaaaaatt ccaaagtaaa tttggtatag
     1141 ataacctgat ggaaacaaat gttgcagagc agtttagcag ggaactaacg gaccttgaaa
     1201 aagatttagc agagataatg aattcattga cacagcactt tgataagaca ttgctgttac
     1261 aagataaaaa aattgataac gatgaacgtg aggagctgtt taaggtggta caaggcgacg
     1321 acaaagaact atacaacatt ttcaaaactc tgcatgaggt aattgatgac gtggacaaaa
     1381 caattcttaa cttgggtcaa tttttgcagg caaaaataaa ggaaaagaca gaactacaca
     1441 gcgaagtttc tgaaataata aacgatttca atagaaattt ggaatatcta ttaatcttta
     1501 aagatatttc caatctgatt gatagcttca aaaattcctg tacacaagat attcaaacaa
     1561 ctaaggaact ttgtgaattt tatgataatt ttgaagaaag ctacggtaac ttagttctag
     1621 aagcaaagag gagaaaggat gtggcaaaca gaatgaaaac tatattgaaa gattgtgaaa
     1681 agcagttgca gaatttagat gctcaggacc aggaagaacg tcagaatttt atagcggaaa
     1741 atggaactta tcttcctgag acaatctggc ccggtaaaat tgacgatttt tcttccctgt
     1801 acactttaaa ttacaacgtg aagaatcctt aggtcgacct cgagtcatgt aattagttat
     1861 gtcacgctta cattcacgcc ctccccccac atccgctcta accgaaaagg aaggagttag
     1921 acaacctgaa gtctaggtcc ctatttattt ttttatagtt atgttagtat taagaacgtt
     1981 atttatattt caaatttttc ttttttttct gtacagacgc gtgtacgcat gtaacattat
     2041 actgaaaacc ttgcttgaga aggttttggg acgctcgaag gctttaattt gcggccggta
     2101 cccaattcgc cctatagtga gtcgtattac gcgcgctcac tggccgtcgt tttacaacgt
     2161 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     2221 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     2281 ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2341 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2401 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2461 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2521 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2581 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2641 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2701 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
     2761 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtaat aactgatata
     2821 attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt
     2881 tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt
     2941 caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt
     3001 cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct
     3061 tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac
     3121 ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa
     3181 cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca
     3241 tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa
     3301 tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata
     3361 cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga
     3421 agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg
     3481 aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt
     3541 caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt
     3601 gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac
     3661 gttccttata tgtagctttc gacatgattt atcttcgttt cctgcaggtt tttgttctgt
     3721 gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat
     3781 ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat
     3841 caaaaaaatt tcaaggaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga
     3901 aaaggtggta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3961 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4021 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4081 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4141 gataataatg gtttcttaga cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt
     4201 taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt tttgtatttg
     4261 gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac
     4321 aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa
     4381 agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt
     4441 tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag
     4501 gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg
     4561 gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt aatttatata
     4621 tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggacccagg
     4681 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     4741 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4801 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4861 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4921 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4981 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5041 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5101 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5161 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5221 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5281 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5341 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5401 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5461 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5521 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5581 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     5641 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     5701 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5761 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5821 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5881 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5941 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6001 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6061 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6121 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6181 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6241 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6301 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6361 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6421 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6481 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     6541 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tg
//
