LOCUS       sequence-240337-        5846 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing S. pombe HA-Atg101.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5846)
  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 5846)
  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-SpAtg101(414)
FEATURES             Location/Qualifiers
     source          1..5846
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    89..110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(125..130,131..148,149..155)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    163..179
                     /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     168..190
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     187..203
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     187..203
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     222..242
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        224..242
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        261..495
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             506..532
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             536..562
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             566..592
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     terminator      1160..1407
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1176..1194)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1425..1444)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1426..1444)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1454..1471)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1454..1470)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1463..1485)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1611..2066
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1698..1717)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1908..1929
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2169..2843)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(2844..3124)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3145..3164)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3264..3286
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3324..3342)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3383..3886
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        3923..4027
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4028..4096,4097..4888)
                     /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(4246..4265)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5059..5647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5548..5567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5801..5818
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
       61 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
      121 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
      181 ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac cctcactaaa
      241 gggaacaaaa gctggagctc ctagttagaa aaagacattt ttgctgtcag tcactgtcaa
      301 gagattcttt tgctggcatt tcttctagaa gcaaaaagag cgatgcgtct tttccgctga
      361 accgttccag caaaaaagac taccaacgca atatggattg tcagaatcat ataaaagaga
      421 agcaaataac tccttgtctt gtatcaattg cattataata tcttcttgtt agtgcaatat
      481 catatagaag tcatcgacta gtatgtaccc atacgatgtt cctgactatg cgggctatcc
      541 ctatgacgtc ccggactatg caggatatcc atatgacgtt ccagattacg ctcccgggat
      601 gacaaacacg gtgacaattg aactgaaaat tggttataag tatgctgctg aggttgtaaa
      661 agccgtttta ggtgtaattc tatttcatag acagttttct actgttcctg ctaggacaat
      721 tgacgtttta gacattacgg ttcctactct ggttggtgca gaactaaatg aacaactagc
      781 aactaaagcg gcggaattta ttgataccat tcgaaatgag gctggtgcaa atggtcagat
      841 gattttactg ttgtatgaaa gatcgcctaa aaagtcttgg tttgggaagg gaaatacaat
      901 tccttgggaa caatggatct tgcacactac cattttagaa gagggtgact cttatcaaga
      961 atcttcttta tctctagagg cagcagttga gcaaattgtg caagcagtta accttcgaag
     1021 cctttcatac ctgcccccag ttgctatgga ttccgggaat tacccgtatg aaattgtaac
     1081 tcctacttca acagaaggtt ggggctcact gctgaagcga atgattatag aaaacgttag
     1141 tggaggtgat taactcgagt catgtaatta gttatgtcac gcttacattc acgccctccc
     1201 cccacatccg ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt
     1261 tattttttta tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt
     1321 tttctgtaca gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt
     1381 ttgggacgct cgaaggcttt aatttgcggc cggtacccaa ttcgccctat agtgagtcgt
     1441 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     1501 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     1561 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg acgcgccctg
     1621 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     1681 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     1741 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1801 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1861 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1921 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     1981 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2041 taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac gcatctgtgc
     2101 ggtatttcac accgcataga tcggcaagtg cacaaacaat acttaaataa atactactca
     2161 gtaataacct atttcttagc atttttgacg aaatttgcta ttttgttaga gtcttttaca
     2221 ccatttgtct ccacacctcc gcttacatca acaccaataa cgccatttaa tctaagcgca
     2281 tcaccaacat tttctggcgt cagtccacca gctaacataa aatgtaagct ttcggggctc
     2341 tcttgccttc caacccagtc agaaatcgag ttccaatcca aaagttcacc tgtcccacct
     2401 gcttctgaat caaacaaggg aataaacgaa tgaggtttct gtgaagctgc actgagtagt
     2461 atgttgcagt cttttggaaa tacgagtctt ttaataactg gcaaaccgag gaactcttgg
     2521 tattcttgcc acgactcatc tccatgcagt tggacgatat caatgccgta atcattgacc
     2581 agagccaaaa catcctcctt aggttgatta cgaaacacgc caaccaagta tttcggagtg
     2641 cctgaactat ttttatatgc ttttacaaga cttgaaattt tccttgcaat aaccgggtca
     2701 attgttctct ttctattggg cacacatata atacccagca agtcagcatc ggaatctaga
     2761 gcacattctg cggcctctgt gctctgcaag ccgcaaactt tcaccaatgg accagaacta
     2821 cctgtgaaat taataacaga catactccaa gctgcctttg tgtgcttaat cacgtatact
     2881 cacgtgctca atagtcacca atgccctccc tcttggccct ctccttttct tttttcgacc
     2941 gaattaattc ttaatcggca aaaaaagaaa agctccggat caagattgta cgtaaggtga
     3001 caagctattt ttcaataaag aatatcttcc actactgcca tctggcgtca taactgcaaa
     3061 gtacacatat attacgatgc tgttctatta aatgcttcct atattatata tatagtaatg
     3121 tcgtgatcta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3181 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3241 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3301 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3361 gataataatg gtttcttaga cggatcgctt gcctgtaact tacacgcgcc tcgtatcttt
     3421 taatgatgga ataatttggg aatttactct gtgtttattt atttttatgt tttgtatttg
     3481 gattttagaa agtaaataaa gaaggtagaa gagttacgga atgaagaaaa aaaaataaac
     3541 aaaggtttaa aaaatttcaa caaaaagcgt actttacata tatatttatt agacaagaaa
     3601 agcagattaa atagatatac attcgattaa cgataagtaa aatgtaaaat cacaggattt
     3661 tcgtgtgtgg tcttctacac agacaagatg aaacaattcg gcattaatac ctgagagcag
     3721 gaagagcaag ataaaaggta gtatttgttg gcgatccccc tagagtcttt tacatcttcg
     3781 gaaaacaaaa actatttttt ctttaatttc tttttttact ttctattttt aatttatata
     3841 tttatattaa aaaatttaaa ttataattat ttttatagca cgtgatgaaa aggacccagg
     3901 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3961 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     4021 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     4081 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     4141 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     4201 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     4261 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     4321 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     4381 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     4441 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     4501 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     4561 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     4621 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     4681 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     4741 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4801 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4861 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     4921 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     4981 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5041 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5101 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5161 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5221 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5281 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5341 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5401 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5461 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5521 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5581 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5641 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5701 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     5761 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     5821 agcggaagag cgcccaatac gcaaac
//
