LOCUS       sequence-459168-        5487 bp    DNA     circular SYN 15-APR-2025
DEFINITION  HA-tagged MYC over-expession construct with point mutations 
            corresponding to the deletion of degron 1 and 2 for transient 
            expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5487)
  AUTHORS   Mark KG, Kolla S, Aguirre JD, Garshott DM, Schmitt S, Haakonsen DL, 
            Xu C, Kater L, Kempf G, Martinez-Gonzalez B, Akopian D, See SK, 
            Thoma NH, Rape M
  TITLE     Orphan quality control shapes network dynamics and gene expression.
  JOURNAL   Cell. 2023 Aug 3;186(16):3460-3475.e23. doi: 
            10.1016/j.cell.2023.06.015. Epub 2023 Jul 20.
  PUBMED    37478862
REFERENCE   2  (bases 1 to 5487)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2023.06.015"; journalName: "Cell"; date: "2023-08-3-
            3"; volume: "186"; issue: "16"; pages: "3460-3475.e23"
COMMENT     Sequence Label: pCS2-HA-Myc-Delta-deg1∆deg2
FEATURES             Location/Qualifiers
     source          1..5487
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..150
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     132..149
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             188..268
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     CDS             278..1597
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
                     SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
                     LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
                     PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
                     LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
                     PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
                     KCTSPRSSDTEENVEEETHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
                     TAAAAAAQAEEQKLISEEDLLRKRREQLKHKLEQERNSEA"
     polyA_signal    1636..1770
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1660..1679)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1714..1733
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1883..1903)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1883..1901)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1922..1938)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1922..1938)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1935..1957)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1946..1962
                     /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)."
     promoter        complement(join(1970..1976,1977..1994,1995..2000))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2015..2036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2153..2170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2324..2912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2404..2423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3083..3874,3875..3943))
                     /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     3706..3725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3944..4048)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4074..4529)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4211..4232)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4423..4442
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(4600..5487,1..97)
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta ggcgtgccta
       61 atgggaggtc tatataagca atgctcgttt agggaaccgc cattctgcct ggggacgtcg
      121 gagcaagctt gatttaggtg acactataga atacaagcta cttgttcttt ttgcaggatc
      181 caccatgtac ccatacgatg ttccagatta cgcttatcct tatgacgtac ctgactatgc
      241 atacccttat gatgtaccag actacgctgg ccggcccatg cccctcaacg ttagcttcac
      301 caacaggaac tatgacctcg actacgactc ggtgcagccg tatttctact gcgacgagga
      361 ggagaacttc taccagcagc agcagcagag cgagctgcag cccccggcgc ccagcgagga
      421 tatctggaag aaattcgagc tgctgcccac cccgcccctg tcccctagcc gccgctccgg
      481 gctctgctcg ccctcctacg ttgcggtcac acccttctcc cttcggggag acaacgacgg
      541 cggtggcggg agcttctcca cggccgacca gctggagatg gtgaccgagc tgctgggagg
      601 agacatggtg aaccagagtt tcatctgcga cccggacgac gagaccttca tcaaaaacat
      661 catcatccag gactgtatgt ggagcggctt ctcggccgcc gccaagctcg tctcagagaa
      721 gctggcctcc taccaggctg cgcgcaaaga cagcggcagc ccgaaccccg cccgcggcca
      781 cagcgtctgc tccacctcca gcttgtacct gcaggatctg agcgccgccg cctcagagtg
      841 catcgacccc tcggtggtct tcccctaccc tctcaacgac agcagctcgc ccaagtcctg
      901 cgcctcgcaa gactccagcg ccttctctcc gtcctcggat tctctgctct cctcgacgga
      961 gtcctccccg cagggcagcc ccgagcccct ggtgctccat gaggagacac cgcccaccac
     1021 cagcagcgac tctgaggagg aacaagaaga tgaggaagaa atcgatgttg tttctgtgga
     1081 aaagaggcag gctcctggca aaaggtcaga gtctggatca ccttctgctg gaggccacag
     1141 caaacctcct cacagcccac tggtcctcaa gaggtgccac gtctccacac atcagcacaa
     1201 ctacgcagcg cctccctcca ctcggaagga ctatcctgct gccaagaggg tcaagttgga
     1261 cagtgtcaga gtcctgagac agatcagcaa caaccgaaaa tgcaccagcc ccaggtcctc
     1321 ggacaccgag gagaatgtcg aggaggagac acacaacgtc ttggagcgcc agaggaggaa
     1381 cgagctaaaa cggagctttt ttgccctgcg tgaccagatc ccggagttgg aaaacaatga
     1441 aaaggccccc aaggtagtta tccttaaaaa agccacagca gccgccgcgg ccgcccaagc
     1501 agaggagcaa aagctcattt ctgaagagga cttgttgcgg aaacgacgag aacagttgaa
     1561 acacaaactt gaacaggaac ggaactctga agcgtaaagg cgcgcctcta gaactatagt
     1621 gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca
     1681 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     1741 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     1801 caggttcagg gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg
     1861 gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     1921 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1981 ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     2041 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     2101 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2161 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2221 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2281 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2341 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2401 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2461 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2521 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2581 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2641 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2701 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2761 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2821 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2881 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2941 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     3001 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     3061 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3121 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3181 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3241 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3301 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3361 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3421 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3481 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3541 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3601 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3661 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3721 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3781 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3841 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3901 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3961 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     4021 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     4081 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     4141 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     4201 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     4261 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     4321 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     4381 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4441 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4501 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
     4561 ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgaccgcca attcaatatg
     4621 gcgtatatgg actcatgcca attcaatatg gtggatctgg acctgtgcca attcaatatg
     4681 gcgtatatgg actcgtgcca attcaatatg gtggatctgg accccagcca attcaatatg
     4741 gcggacttgg caccatgcca attcaatatg gcggacctgg cactgtgcca actggggagg
     4801 ggtctacttg gcacggtgcc aagtttgagg aggggtcttg gccctgtgcc aagtccgcca
     4861 tattgaattg gcatggtgcc aataatggcg gccatattgg ctatatgcca ggatcaatat
     4921 ataggcaata tccaatatgg ccctatgcca atatggctat tggccaggtt caatactatg
     4981 tattggccct atgccatata gtattccata tatgggtttt cctattgacg tagatagccc
     5041 ctcccaatgg gcggtcccat ataccatata tggggcttcc taataccgcc catagccact
     5101 cccccattga cgtcaatggt ctctatatat ggtctttcct attgacgtca tatgggcggt
     5161 cctattgacg tatatggcgc ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg
     5221 ctcaatgccc attgacgtca ataggaccac ccaccattga cgtcaatggg atggctcatt
     5281 gcccattcat atccgttctc acgcccccta ttgacgtcaa tgacggtaaa tggcccactt
     5341 ggcagtacat caatatctat taatagtaac ttggcaagta cattactatt ggaagtacgc
     5401 cagggtacat tggcagtact cccattgacg tcaatggcgg taaatggccc gcgatggctg
     5461 ccaagtacat ccccattgac gtcaatg
//
