LOCUS       sequence-240339-        6622 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing S. pombe HA-Atg17.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6622)
  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 6622)
  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: pCuHA-SpAtg17(416)
FEATURES             Location/Qualifiers
     source          1..6622
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     12..29
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    146..167
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(182..187,188..205,206..212)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    220..236
                     /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     225..247
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     244..260
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     244..260
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     279..299
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        281..299
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        318..552
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             563..589
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             593..619
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             623..649
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      1898..2145
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1914..1932)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(2163..2182)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2164..2182)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2192..2209)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2192..2208)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2201..2223)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2349..2804
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2436..2455)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2646..2667
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2935..3738)
                     /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(3739..3954)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(3978..3997)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4097..4119
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4157..4175)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4216..4719
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4756..4860
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4861..4929,4930..5721)
                     /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(5079..5098)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5892..6480
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6381..6400
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
       61 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
      121 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
      181 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
      241 acacaggaaa cagctatgac catgattacg ccaagcgcgc aattaaccct cactaaaggg
      301 aacaaaagct ggagctccta gttagaaaaa gacatttttg ctgtcagtca ctgtcaagag
      361 attcttttgc tggcatttct tctagaagca aaaagagcga tgcgtctttt ccgctgaacc
      421 gttccagcaa aaaagactac caacgcaata tggattgtca gaatcatata aaagagaagc
      481 aaataactcc ttgtcttgta tcaattgcat tataatatct tcttgttagt gcaatatcat
      541 atagaagtca tcgactagta tgtacccata cgatgttcct gactatgcgg gctatcccta
      601 tgacgtcccg gactatgcag gatatccata tgacgttcca gattacgctc ccgggatgga
      661 attgttgcag cagtggactc agcaagccaa agcagctctt actcaagcca ggcaactgtg
      721 tggtgatgca cataagttta atgaagatgc aaaaactgat ttgagaaatt ccataaagca
      781 gcatcaacag ctaaaagagt tggcgaagct cacagcatct caatgtactc gtttagactc
      841 ttcaacagct cttattaagc agttattaga tctagttcaa aattatccaa catttaatca
      901 attaaacgtc cttcatgatc gattggaatc gtcacttaaa cgtcttcgtg actgtacatt
      961 agatccagcc ttaggaagcg aatataccaa tctttatgca tttgtagatg atactgcatt
     1021 agaagatttg aaaaccaggc tccgaggcgt aactgatggt gtttggaatg ctttcgaaaa
     1081 gctagcgggt ttactggaag aagatttgtg tgccaattat cataaacgtt tagaagctgt
     1141 ttcattagac ttcttacctc ccgcttataa cgatactgcc gaagaacttg cggatttgct
     1201 tttgcaagtt gctcaacatt acgatcaatg ctccgaagct ctcaatattt atgacaccct
     1261 ttctgatgcg gagaagaaag atctccaaga agtgcttcaa tcagactcta atcatgttcc
     1321 ttcggtattg acagaactta ggagtggatt agatcaaaca attcattatt tcaacgcagt
     1381 tcaaagttat aagtcaaagg tagattcggc aacatcgatt ttggaagcct tggctgagga
     1441 gttaaacaaa aaccagttga ctaatcaaag gcatgaagcc gcgcatgagt taatgagggc
     1501 gcaaaccggc ctggagattc cgcaattagc tcaagaactc gtgcagcttg aaagacatta
     1561 cacacatttt gctaaagcgt ataccgctct ccttcaggaa attcacagac gacaaaccta
     1621 tgaaaattgt gtaagatcca ttgtggacga atttgtggga cgcctggaga aggaacaaca
     1681 ggcggaggcg aaatgtcgta tcgatttttt caatcaatat ggtgattatt taccccagac
     1741 tctatgggga gctgtaacag atcctccttt acattttgaa attatagagc accaatatac
     1801 agaattgcca aacgtaaagg taattcctga taaaaacgat aagaaatcga agcaacgtga
     1861 aaaatcttct actacagcct ctaagcgatg actcgagtca tgtaattagt tatgtcacgc
     1921 ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
     1981 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
     2041 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
     2101 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcggccg gtacccaatt
     2161 cgccctatag tgagtcgtat tacgcgcgct cactggccgt cgttttacaa cgtcgtgact
     2221 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     2281 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     2341 gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     2401 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2461 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2521 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2581 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2641 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2701 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2761 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt
     2821 ctccttacgc atctgtgcgg tatttcacac cgcatagggt aataactgat ataattaaat
     2881 tgaagctcta atttgtgagt ttagtataca tgcatttact tataatacag ttttttagtt
     2941 ttgctggccg catcttctca aatatgcttc ccagcctgct tttctgtaac gttcaccctc
     3001 taccttagca tcccttccct ttgcaaatag tcctcttcca acaataataa tgtcagatcc
     3061 tgtagagacc acatcatcca cggttctata ctgttgaccc aatgcgtctc ccttgtcatc
     3121 taaacccaca ccgggtgtca taatcaacca atcgtaacct tcatctcttc cacccatgtc
     3181 tctttgagca ataaagccga taacaaaatc tttgtcgctc ttcgcaatgt caacagtacc
     3241 cttagtatat tctccagtag atagggagcc cttgcatgac aattctgcta acatcaaaag
     3301 gcctctaggt tcctttgtta cttcttctgc cgcctgcttc aaaccgctaa caatacctgg
     3361 gcccaccaca ccgtgtgcat tcgtaatgtc tgcccattct gctattctgt atacacccgc
     3421 agagtactgc aatttgactg tattaccaat gtcagcaaat tttctgtctt cgaagagtaa
     3481 aaaattgtac ttggcggata atgcctttag cggcttaact gtgccctcca tggaaaaatc
     3541 agtcaagata tccacatgtg tttttagtaa acaaattttg ggacctaatg cttcaactaa
     3601 ctccagtaat tccttggtgg tacgaacatc caatgaagca cacaagtttg tttgcttttc
     3661 gtgcatgata ttaaatagct tggcagcaac aggactagga tgagtagcag cacgttcctt
     3721 atatgtagct ttcgacatga tttatcttcg tttcctgcag gtttttgttc tgtgcagttg
     3781 ggttaagaat actgggcaat ttcatgtttc ttcaacacta catatgcgta tatataccaa
     3841 tctaagtctg tgctccttcc ttcgttcttc cttctgttcg gagattaccg aatcaaaaaa
     3901 atttcaagga aaccgaaatc aaaaaaaaga ataaaaaaaa aatgatgaat tgaaaaggtg
     3961 gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
     4021 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     4081 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     4141 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
     4201 atggtttctt agacggatcg cttgcctgta acttacacgc gcctcgtatc ttttaatgat
     4261 ggaataattt gggaatttac tctgtgttta tttattttta tgttttgtat ttggatttta
     4321 gaaagtaaat aaagaaggta gaagagttac ggaatgaaga aaaaaaaata aacaaaggtt
     4381 taaaaaattt caacaaaaag cgtactttac atatatattt attagacaag aaaagcagat
     4441 taaatagata tacattcgat taacgataag taaaatgtaa aatcacagga ttttcgtgtg
     4501 tggtcttcta cacagacaag atgaaacaat tcggcattaa tacctgagag caggaagagc
     4561 aagataaaag gtagtatttg ttggcgatcc ccctagagtc ttttacatct tcggaaaaca
     4621 aaaactattt tttctttaat ttcttttttt actttctatt tttaatttat atatttatat
     4681 taaaaaattt aaattataat tatttttata gcacgtgatg aaaaggaccc aggtggcact
     4741 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     4801 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     4861 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4921 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     4981 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5041 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5101 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5161 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5221 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5281 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5341 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5401 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5461 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5521 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     5581 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     5641 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5701 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     5761 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5821 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5881 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5941 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6001 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6061 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6121 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6181 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6241 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6301 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6361 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6421 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6481 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6541 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     6601 gataccgctc gccgcagccg aa
//
