LOCUS       sequence-202801-        7767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses C terminally 3xFLAG tagged ZNF598 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7767)
  AUTHORS   Juszkiewicz S, Hegde RS
  TITLE     Initiation of Quality Control during Poly(A) Translation Requires 
            Site-Specific Ribosome Ubiquitination.
  JOURNAL   Mol Cell. 2017 Feb 16;65(4):743-750.e4. doi: 
            10.1016/j.molcel.2016.11.039. Epub 2017 Jan 5.
  PUBMED    28065601
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"; doi: 
            "10.1016/j.molcel.2016.11.039"; journalName: "Mol Cell"; date: 
            "2017-02-16- 16"; volume: "65"; issue: "4"; pages: "743-750.e4"
COMMENT     Sequence Label: pcDNA3.1-ZNF598-TEV-3xFLAG
FEATURES             Location/Qualifiers
     source          1..7767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      693..702
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3413..3433
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3437..3502
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(3537..3554)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3543..3767
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3813..4241
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3900..3919)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4110..4131
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4250..4270)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4255..4584
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4435..4570
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4497..4516
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4632..4679
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4698..5072
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    5202..5323
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5239..5258)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5293..5312
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5372..5388)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5372..5388)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5385..5407)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5396..5412
                     /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(5420..5426,5427..5444,5445..5450))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5465..5486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5603..5620)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5774..6362)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5854..5873)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6533..7324,7325..7393))
                     /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     7156..7175
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7394..7498)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7577..7596)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgtcg acagatctcc cgggaattca aaccatggcg gcggcggggg gcgccgaggg
      721 gcggcgcgcg gccctggagg cggcggcggc ggcagctcct gagcggggag gcgggagctg
      781 cgtgctgtgc tgcggagacc tggaggccac ggcgctgggc cgctgcgacc acccggtgtg
      841 ctaccgctgc tctaccaaga tgcgggtgct ctgcgagcag cgctactgcg ccgtgtgccg
      901 cgaggagctg cgccaggtgg tctttgggaa gaagcttcct gcctttgcca ccatccccat
      961 ccaccagctg cagcatgaga agaaatatga tatctacttt gcagatggaa aggtgtacgc
     1021 attgtacagg cagctgctgc agcacgagtg cccgcggtgc cccgagctgc cacctttcag
     1081 cctcttcggg gacctggagc agcacatgcg gaggcagcat gagctcttct gctgccgtct
     1141 gtgcctccag cacctccaga tcttcacata tgagcgcaag tggtactcgc gcaaggacct
     1201 ggcccggcat cgcatgcagg gtgaccccga tgacacgtcg caccgtgggc acccgctctg
     1261 caagttctgt gacgagcgct acctggacaa tgatgagctg cttaagcacc tgcgccgcga
     1321 ccactacttc tgccacttct gcgactcgga cggggcccag gactactaca gcgactatgc
     1381 ctacctgcgt gagcacttcc gggagaagca ctttctgtgt gaggaaggcc gctgcagcac
     1441 ggagcagttc acccacgcct tccgcaccga gatcgacctc aaggcccaca ggacggcctg
     1501 ccacagtcgc agccgcgccg aggcacgcca gaaccgccac atcgacctgc agttcagcta
     1561 cgcgccacgg cactcgcgcc ggaacgaggg ggtcgttggt ggcgaagact acgaggaggt
     1621 ggacaggtac agccgccagg gccgagtggc ccgggctggc actcgcggag cccagcagag
     1681 ccgccgagga agctggaggt acaaaaggga agaagaggac cgagaagtag cagctgctgt
     1741 ccgggcctcc gtggccgcac agcagcagga ggaggctcgc aggagtgagg atcaggagga
     1801 aggcggtagg cccaagaagg aggaggcagc ggcgcgggga cctgaggatc cccgtggccc
     1861 ccggcgctca ccccggactc agggcgaagg cccaggcccc aaggaaacct cgacaaatgg
     1921 tcctgtaagc caagaagcct tctcggtgac aggcccagcc gccccagggt gtgtgggggt
     1981 gccaggcgcc ctcccaccac ccagcccgaa gctcaaggac gaagacttcc ccagcctctc
     2041 tgcctccact tcctcctcct gctccactgc agcaaccccg ggccctgtgg ggttggcgct
     2101 gccgtacgcc atccctgcca gaggcaggag tgccttccag gaggaggact tccccgccct
     2161 ggtgtcctcg gtgcccaagc ctggcaccgc ccccaccagc cttgtctctg cctggaacag
     2221 cagcagtagc agcaagaagg tagcacagcc cccactctcg gcgcaggcta ccggcagcgg
     2281 ccagcccacc aggaaggctg ggaaggggag caggggcggc aggaagggcg gcccgccctt
     2341 cacacaggag gaggaggagg acggcggccc ggccctgcag gagcttctga gcacacgccc
     2401 cacgggctcc gtctcctcca cactggggct ggcctccatc caaccctcta aagttgggaa
     2461 gaagaagaaa gtgggctcgg agaagccagg caccacattg ccacagcccc cgcccgctac
     2521 ctgtccccca ggggctttgc aggccccgga agctcctgcc agcagagccg aggggccagt
     2581 tgccgtcgtc gttaatggac acacggaggg cccggcccct gctcggagtg cccccaagga
     2641 accccctggg ctcccaaggc ccctggggtc cttcccctgc cccacgccac aggaggactt
     2701 cccagcgctc ggcggcccct gcccaccccg gatgccgccg cccccaggct tcagcgctgt
     2761 ggtgctcctg aagggcacgc ctcccccacc cccgccgggc ctggtgcccc caatcagcaa
     2821 gccgccccct ggcttctctg gccttctgcc tagcccccac ccggcctgtg tccccagccc
     2881 cgccaccacc accaccacaa aagcacccag gctgctgcct gccccacggg cgtacctagt
     2941 ccccgagaac ttccgggaga ggaaccttca gctcatccag tccatcaggg acttcctgca
     3001 gagcgacgag gcccgcttca gcgagttcaa gagccactca ggggagttca gacagggcct
     3061 gatctccgca gcccagtatt acaagagttg ccgggacctg ctgggggaga atttccagaa
     3121 ggtctttaat gagctgctgg tcctgctgcc cgacacggcc aagcagcagg agctcctgtc
     3181 tgcacacacg gacttctgca accgcgagaa gcctctgagc accaagtcca agaagaacaa
     3241 gaagagcgcg tggcaggcca ccacccagca ggcgggcctg gactgccgtg tgtgccccac
     3301 ctgccagcag gtgctcgcgc atggcgatgc cagcagccac caggcgctgc atgctgcccg
     3361 ggacgacgac ttcccctccc tgcaagccat cgccaggatc atcacgtcta gagagaacct
     3421 gtactttcag ggatctgact acaaagacca tgacggtgat tataaagatc atgacatcga
     3481 ttacaaggat gacgatgaca agtaacctag agggcccgtt taaacccgct gatcagcctc
     3541 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     3601 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     3661 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     3721 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     3781 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     3841 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3901 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3961 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4021 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4081 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4141 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     4201 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     4261 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     4321 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     4381 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     4441 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     4501 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     4561 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     4621 gatcagcacg tgttgacaat taatcatcgg catagtatat cggcatagta taatacgaca
     4681 aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt tccggtgctc accgcgcgcg
     4741 acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg gacttcgtgg
     4801 aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc gcggtccagg
     4861 accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg gacgagctgt
     4921 acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg ccggccatga
     4981 ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg gccggcaact
     5041 gcgtgcactt cgtggccgag gagcaggact gacacgtgct acgagatttc gattccaccg
     5101 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     5161 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     5221 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5281 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     5341 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5401 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5461 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5521 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5581 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5641 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5701 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5761 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5821 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5881 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5941 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6001 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6061 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6181 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6421 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6601 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6661 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6721 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6781 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6841 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6901 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6961 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7021 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7081 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7141 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7201 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7261 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7321 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7381 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7441 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7501 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcgatct cccgatcccc
     7561 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc
     7621 ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc
     7681 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
     7741 gcgatgtacg ggccagatat acgcgtt
//
