LOCUS       sequence-423110-        7767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of 4Myc-Pho8 under CPU1 promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7767)
  AUTHORS   He CW, Cui XF, Ma SJ, Xu Q, Ran YP, Chen WZ, Mu JX, Li H, Zhu J, 
            Gong Q, Xie Z
  TITLE     Membrane recruitment of Atg8 by Hfl1 facilitates turnover of 
            vacuolar membrane proteins in yeast cells approaching stationary 
            phase.
  JOURNAL   BMC Biol. 2021 Jun 4;19(1):117. doi: 10.1186/s12915-021-01048-7.
  PUBMED    34088313
REFERENCE   2  (bases 1 to 7767)
  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: "BMC Biol.";
            date: "2021-06-4"; volume: "19(1):117. doi"; pages: " 
            10.1186/s12915-021-01048-7"
COMMENT     Sequence Label: BS-Trp-PCUP1-4myc-Pho8
FEATURES             Location/Qualifiers
     source          1..7767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..21
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5..21
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     40..60
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        42..60
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        796..1073
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     1810..1826
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     1811..1827
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1831..2065
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             2121..2150
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2157..2186
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2193..2222
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2229..2258
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4997..5016)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4998..5016)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5023..5040)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5023..5039)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5032..5054)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5181..5636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5268..5287)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5478..5499
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5662..5766
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5767..5835,5836..6627)
                     /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(5985..6004)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6798..7386
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7287..7306
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7540..7557
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7674..7695
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7710..7715,7716..7733,7734..7740)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7748..7764
                     /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     join(7753..7767,1..8)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc tcactaaagg
       61 gaacaaaagc tggtactatt agctgaattg ccactgctat cgttgttagt ggcgttagtg
      121 cttgcattca aagacatgga gggcgttatt acgccggagc tcctcgacag cagatctgat
      181 gactggtcaa tatatttttg cattgaggct ctgtttggaa ttatattttg agatgaccca
      241 tctaatgtac tggtatcacc agatttcatg tcgtttttta aagcggctgc ttgagtctta
      301 gcaatagcgt caccatctgg tgaatccttt gaaggaacca ctgacgaagg tttggacagt
      361 gacgaagagg atctttcctg ctttgaatta gtcgcgctgg gagcagatga cgagttggtg
      421 gagctggggg caggattgct ggccgtcgtg ggtcctgaat gggtccttgg ctggtccatc
      481 tctattctga aaacggaaga ggagtaggga atattactgg ctgaaaataa gtcttgaatg
      541 aacgtatacg cgtatatttc taccaatctc tcaacactga gtaatggtag ttataagaaa
      601 gagaccgagt tagggacagt tagaggcggt ggagatattc cttatggcat gtctggcgat
      661 gataaaactt ttcaaacggc agccccgatc taaaagagct gacagggaaa tggtcagaaa
      721 aagaaacgtg cacccgcccg tctggacgcg ccgctcaccc gcacggcaga gaccaatcag
      781 taaaaatcaa cggttaacga cattactata tatataatat aggaagcatt taatagaaca
      841 gcatcgtaat atatgtgtac tttgcagtta tgacgccaga tggcagtagt ggaagatatt
      901 ctttattgaa aaatagcttg tcaccttacg tacaatcttg atccggagct tttctttttt
      961 tgccgattaa gaattcggtc gaaaaaagaa aaggagaggg ccaagaggga gggcattggt
     1021 gactattgag cacgtgagta tacgtgatta agcacacaaa ggcagcttgg agtatgctcg
     1081 ttaaagtgtg tggtttgcaa accgttgaag ctgcaaagac tgctgtggat gatggtgctg
     1141 attacttagg tatcatttgt gttcccggta ggaaaagaac cattgactca tctgttgcga
     1201 aaggtatttc aactgcagtt caccaacaag agaacgtgaa aggtactaaa ctagtcgggg
     1261 tgtttagaaa tcagtccgtt gatgatgtcc ttcaactgta ccacgaatat aatctagatg
     1321 tgatacaatt acatggagat gaagatatta aagaatacag atctttgatc ccatcttcaa
     1381 ttccaatcat taagaggttc cagttcccac aggattgtga attactactg gacctgtatg
     1441 aacacgtaga caatgtgctg acgttgttcg attctggtga aggtggcact ggtgagaaat
     1501 tgaattggag tgcaatttcc agttggtctg caagtcatcc cgagataaaa ttcattatcg
     1561 ctggtggatt gaatcctgat aacgtttctg ttgccattaa tatgttacca aatgcgatcg
     1621 gtgtcgatgt aagtggagga gtagagactg atggtatcaa ggatttagaa aaggtaaagc
     1681 tattcatcca gcaggcctct caatagagga aaaaacgtat ctttcttcaa ttacacatac
     1741 gtatatataa attgatatat atatagtgaa tgatcgttcc ccggccgctc tagggtaccg
     1801 ggccccccct cgaggtcgac ggtatcgata ctagttagaa aaagacattt ttgctgtcag
     1861 tcactgtcaa gagattcttt tgctggcatt tcttctagaa gcaaaaagag cgatgcgtct
     1921 tttccgctga accgttccag caaaaaagac taccaacgca atatggattg tcagaatcat
     1981 ataaaagaga agcaaataac tccttgtctt gtatcaattg cattataata tcttcttgtt
     2041 agtgcaatat catatagaag tcatcagctt cgtacgctgc aggtcgtggg attcctgggt
     2101 taattaacat cgctagcatg gaacaaaaat tgatttctga agaagatttg ccaagagaac
     2161 aaaaattgat ttctgaagaa gatttgccta gcgaacaaaa attgatttct gaagaagatt
     2221 tgccaagaga acaaaaattg atttctgaag aagatttgcc taggatgatg actcacacat
     2281 taccaagcga acagacacgt cttgttcctg gatctgactc gagctctcgt cctaaaaaga
     2341 gacggatctc gaagagatca aagataatag tatccactgt ggtctgtatt ggtttgttgc
     2401 tggttttagt gcaactggca tttccaagca gttttgcatt acgttctgca tcacacaaga
     2461 agaagaatgt catattcttc gtgacggatg ggatgggacc tgcgtctttg tccatggcaa
     2521 gatcattcaa ccagcatgtt aatgatttac ctatagacga catcctgaca cttgacgagc
     2581 attttatcgg gtcttcgaga acaaggtcct cggactcact agtcactgac tcagctgcgg
     2641 gcgccaccgc atttgcatgt gcattaaaga gctataatgg ggccatcggt gttgatccgc
     2701 atcacagacc ttgtggtaca gtgctcgagg cggctaagtt agcaggttat ctcactgggc
     2761 ttgttgttac gacaagaatc acagatgcca ctccggcttc attcagttca catgttgatt
     2821 acagatggca ggaagatctt atcgctacgc accagttagg cgaatatcct ctggggagag
     2881 tagtagactt actcatgggt ggcggcagga gccatttcta ccctcaaggc gaaaaggctt
     2941 ctccatatgg tcaccacggt gctagaaaag atggaagaga tttaatcgat gaggctcaat
     3001 ctaatggctg gcagtacgtg ggtgaccgta aaaattttga ttctttgttg aaaagccacg
     3061 gtgaaaacgt tactttacct ttcttgggtc tctttgcaga taacgacatt ccttttgaaa
     3121 ttgacagaga tgaaaaggag tatccttccc ttaaggaaca agtcaaggtg gctttgggtg
     3181 ccttggaaaa agcttccaat gaggacaaag attctaatgg gtttttctta atggtggaag
     3241 gctccagaat tgatcatgct ggtcaccaaa acgatccagc ttcgcaggta agggaggtgt
     3301 tggctttcga tgaagcattc caatacgttt tggagtttgc tgaaaattcc gatacggaga
     3361 ctgttttggt ctctacgtcg gatcatgaaa cgggtggttt agtcacttca agacaagtca
     3421 ctgctagcta tcctcagtat gtctggtatc cacaggtgct cgccaacgcc acccattcag
     3481 gtgagttttt gaaaagaaaa ctagtagatt ttgttcatga gcataagggc gcaagcagca
     3541 aaatagaaaa tttcatcaaa catgagatcc tggaaaaaga tttgggtatt tacgattaca
     3601 cggatagcga tttagaaaca ttaattcatt tggatgacaa cgcaaatgct atccaagaca
     3661 aactgaatga catggtatca tttagggccc aaattggctg gaccacgcat ggtcacagcg
     3721 cagtcgacgt caacatatat gcatacgcca ataagaaagc tacatggtca tatgtcttga
     3781 acaatttaca aggcaatcac gaaaatacgg aagttggcca gttcttagaa aactttttag
     3841 aattgaacct gaacgaagtc actgatttga ttagggacac caaacacaca tcagattttg
     3901 atgctacaga aatagctagt gaagtacaac attatgacga atactaccat gagttgacca
     3961 actgatccta tctaactctc ccgcggtggg ttattactga gtagtattta tttaagtatt
     4021 gtttgtgcac ttgcctgcag gccttttgaa aagcaagcat aaaagatcta aacataaaat
     4081 ctgtaaaata acaagatgta aagataatgc taaatcattt ggctttttga ttgattgtac
     4141 aggaaaatat acatcgcagg gggttgactt ttaccatttc accgcaatgg aatcaaactt
     4201 gttgaagaga atgttcacag gcgcatacgc tacaatgacc cgattcttgc tagccttttc
     4261 tcggtcttgc aaacaaccgc cggcagctta gtatataaat acacatgtac atacctctct
     4321 ccgtatcctc gtaatcattt tcttgtattt atcgtctttt cgctgtaaaa actttatcac
     4381 acttatctca aatacactta ttaaccgctt ttactattat cttctacgct gacagtaata
     4441 tcaaacagtg acacatatta aacacagtgg tttctttgca taaacaccat cagcctcaag
     4501 tcgtcaagta aagatttcgt gttcatgcag atagataaca atctatatgt tgataattag
     4561 cgttgcctca tcaatgcgag atccgtttaa ccggacccta gtgcacttac cccacgttcg
     4621 gtccactgtg tgccgaacat gctccttcac tattttaaca tgtggaattc ttgaaagaat
     4681 gaaatcgcca tgccaagcca tcacacggtc ttttatgcaa ttgattgacc gcctgcaaca
     4741 cataggcagt aaaattttta ctgaaacgta tataatcatc ataagcgaca agtgaggcaa
     4801 cacctttgtt accacattga caaccccagg tattcatact tcctattagc ggaatcagga
     4861 gtgcaaaaag agaaaataaa agtaaaaagg tagggcaaca catagtatga atacaaacgt
     4921 tccaatattc agttctccgg tcagagattt accaaggtct ttcgaacaaa aacatttagc
     4981 ggttgtagac aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca
     5041 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     5101 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     5161 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5221 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5281 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5341 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5401 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5461 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5521 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5581 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
     5641 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5701 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5761 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5821 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5881 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5941 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6001 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     6061 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     6121 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     6181 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6241 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6301 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6361 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6421 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6481 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6541 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6601 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6661 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6721 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6781 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6841 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6901 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6961 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7021 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     7081 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     7141 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     7201 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7261 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7321 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7381 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7441 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     7501 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7561 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7621 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7681 agttagctca ctcattaggc accccaggct ttacacttta tgctcccggc tcgtatgttg
     7741 tgtggaattg tgagcggata acaattt
//
