LOCUS       sequence-240334-        6538 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing S. pombe Atg17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6538)
  AUTHORS   Nanji T, Liu X, Chew LH, Li FK, Biswas M, Yu ZQ, Lu S, Dong MQ, Du 
            LL, Klionsky DJ, Yip CK
  TITLE     Conserved and unique features of the fission yeast core Atg1 
            complex.
  JOURNAL   Autophagy. 2017;13(12):2018-2027. doi: 
            10.1080/15548627.2017.1382782. Epub 2017 Nov 23.
  PUBMED    28976798
REFERENCE   2  (bases 1 to 6538)
  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: 
            "Autophagy."; date: "2017"; volume: "13(12):2018-2027. doi"; pages: 
            " 10.1080/15548627.2017.1382782. Epub 2017 Nov 23"
COMMENT     Sequence Label: pCu-SpAtg17(416)
FEATURES             Location/Qualifiers
     source          1..6538
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    83..104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(119..124,125..142,143..149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    157..173
                     /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     162..184
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     181..197
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     181..197
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     216..236
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        218..236
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        255..489
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     terminator      1751..1998
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1767..1785)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2016..2035)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2017..2035)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2045..2062)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2045..2061)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2054..2076)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2202..2657
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2289..2308)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2499..2520
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2788..3591)
                     /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(3592..3807)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3831..3850)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3950..3972
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4010..4028)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4069..4572
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4609..4713
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4714..4782,4783..5574)
                     /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(4932..4951)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5745..6333
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6234..6253
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6487..6504
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
       61 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      121 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      181 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
      241 aaaagctgga gctcctagtt agaaaaagac atttttgctg tcagtcactg tcaagagatt
      301 cttttgctgg catttcttct agaagcaaaa agagcgatgc gtcttttccg ctgaaccgtt
      361 ccagcaaaaa agactaccaa cgcaatatgg attgtcagaa tcatataaaa gagaagcaaa
      421 taactccttg tcttgtatca attgcattat aatatcttct tgttagtgca atatcatata
      481 gaagtcatcg actagtggat cccccgggat ggaattgttg cagcagtgga ctcagcaagc
      541 caaagcagct cttactcaag ccaggcaact gtgtggtgat gcacataagt ttaatgaaga
      601 tgcaaaaact gatttgagaa attccataaa gcagcatcaa cagctaaaag agttggcgaa
      661 gctcacagca tctcaatgta ctcgtttaga ctcttcaaca gctcttatta agcagttatt
      721 agatctagtt caaaattatc caacatttaa tcaattaaac gtccttcatg atcgattgga
      781 atcgtcactt aaacgtcttc gtgactgtac attagatcca gccttaggaa gcgaatatac
      841 caatctttat gcatttgtag atgatactgc attagaagat ttgaaaacca ggctccgagg
      901 cgtaactgat ggtgtttgga atgctttcga aaagctagcg ggtttactgg aagaagattt
      961 gtgtgccaat tatcataaac gtttagaagc tgtttcatta gacttcttac ctcccgctta
     1021 taacgatact gccgaagaac ttgcggattt gcttttgcaa gttgctcaac attacgatca
     1081 atgctccgaa gctctcaata tttatgacac cctttctgat gcggagaaga aagatctcca
     1141 agaagtgctt caatcagact ctaatcatgt tccttcggta ttgacagaac ttaggagtgg
     1201 attagatcaa acaattcatt atttcaacgc agttcaaagt tataagtcaa aggtagattc
     1261 ggcaacatcg attttggaag ccttggctga ggagttaaac aaaaaccagt tgactaatca
     1321 aaggcatgaa gccgcgcatg agttaatgag ggcgcaaacc ggcctggaga ttccgcaatt
     1381 agctcaagaa ctcgtgcagc ttgaaagaca ttacacacat tttgctaaag cgtataccgc
     1441 tctccttcag gaaattcaca gacgacaaac ctatgaaaat tgtgtaagat ccattgtgga
     1501 cgaatttgtg ggacgcctgg agaaggaaca acaggcggag gcgaaatgtc gtatcgattt
     1561 tttcaatcaa tatggtgatt atttacccca gactctatgg ggagctgtaa cagatcctcc
     1621 tttacatttt gaaattatag agcaccaata tacagaattg ccaaacgtaa aggtaattcc
     1681 tgataaaaac gataagaaat cgaagcaacg tgaaaaatct tctactacag cctctaagcg
     1741 atgactcgag tcatgtaatt agttatgtca cgcttacatt cacgccctcc ccccacatcc
     1801 gctctaaccg aaaaggaagg agttagacaa cctgaagtct aggtccctat ttattttttt
     1861 atagttatgt tagtattaag aacgttattt atatttcaaa tttttctttt ttttctgtac
     1921 agacgcgtgt acgcatgtaa cattatactg aaaaccttgc ttgagaaggt tttgggacgc
     1981 tcgaaggctt taatttgcgg ccggtaccca attcgcccta tagtgagtcg tattacgcgc
     2041 gctcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     2101 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     2161 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct gtagcggcgc
     2221 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2281 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2341 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     2401 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     2461 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     2521 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     2581 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     2641 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     2701 caccgcatag ggtaataact gatataatta aattgaagct ctaatttgtg agtttagtat
     2761 acatgcattt acttataata cagtttttta gttttgctgg ccgcatcttc tcaaatatgc
     2821 ttcccagcct gcttttctgt aacgttcacc ctctacctta gcatcccttc cctttgcaaa
     2881 tagtcctctt ccaacaataa taatgtcaga tcctgtagag accacatcat ccacggttct
     2941 atactgttga cccaatgcgt ctcccttgtc atctaaaccc acaccgggtg tcataatcaa
     3001 ccaatcgtaa ccttcatctc ttccacccat gtctctttga gcaataaagc cgataacaaa
     3061 atctttgtcg ctcttcgcaa tgtcaacagt acccttagta tattctccag tagataggga
     3121 gcccttgcat gacaattctg ctaacatcaa aaggcctcta ggttcctttg ttacttcttc
     3181 tgccgcctgc ttcaaaccgc taacaatacc tgggcccacc acaccgtgtg cattcgtaat
     3241 gtctgcccat tctgctattc tgtatacacc cgcagagtac tgcaatttga ctgtattacc
     3301 aatgtcagca aattttctgt cttcgaagag taaaaaattg tacttggcgg ataatgcctt
     3361 tagcggctta actgtgccct ccatggaaaa atcagtcaag atatccacat gtgtttttag
     3421 taaacaaatt ttgggaccta atgcttcaac taactccagt aattccttgg tggtacgaac
     3481 atccaatgaa gcacacaagt ttgtttgctt ttcgtgcatg atattaaata gcttggcagc
     3541 aacaggacta ggatgagtag cagcacgttc cttatatgta gctttcgaca tgatttatct
     3601 tcgtttcctg caggtttttg ttctgtgcag ttgggttaag aatactgggc aatttcatgt
     3661 ttcttcaaca ctacatatgc gtatatatac caatctaagt ctgtgctcct tccttcgttc
     3721 ttccttctgt tcggagatta ccgaatcaaa aaaatttcaa ggaaaccgaa atcaaaaaaa
     3781 agaataaaaa aaaaatgatg aattgaaaag gtggtatggt gcactctcag tacaatctgc
     3841 tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga
     3901 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     3961 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata
     4021 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgga tcgcttgcct
     4081 gtaacttaca cgcgcctcgt atcttttaat gatggaataa tttgggaatt tactctgtgt
     4141 ttatttattt ttatgttttg tatttggatt ttagaaagta aataaagaag gtagaagagt
     4201 tacggaatga agaaaaaaaa ataaacaaag gtttaaaaaa tttcaacaaa aagcgtactt
     4261 tacatatata tttattagac aagaaaagca gattaaatag atatacattc gattaacgat
     4321 aagtaaaatg taaaatcaca ggattttcgt gtgtggtctt ctacacagac aagatgaaac
     4381 aattcggcat taatacctga gagcaggaag agcaagataa aaggtagtat ttgttggcga
     4441 tccccctaga gtcttttaca tcttcggaaa acaaaaacta ttttttcttt aatttctttt
     4501 tttactttct atttttaatt tatatattta tattaaaaaa tttaaattat aattattttt
     4561 atagcacgtg atgaaaagga cccaggtggc acttttcggg gaaatgtgcg cggaacccct
     4621 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     4681 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     4741 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     4801 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     4861 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     4921 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     4981 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5041 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5101 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5161 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5221 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5281 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5341 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5401 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5461 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5521 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5581 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     5641 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     5701 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     5761 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     5821 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     5881 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     5941 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6001 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6061 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6121 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6181 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6241 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6301 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6361 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6421 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6481 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctc
//
