LOCUS       sequence-342078-        4456 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Purification of His-Lipoyl-hTRIM21 protein from bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4456)
  AUTHORS   Clift D, McEwan WA, Labzin LI, Konieczny V, Mogessie B, James LC, 
            Schuh M
  TITLE     A Method for the Acute and Rapid Degradation of Endogenous Proteins.
  JOURNAL   Cell. 2017 Nov 13. pii: S0092-8674(17)31255-2. doi: 
            10.1016/j.cell.2017.10.033.
  PUBMED    29153837
REFERENCE   2  (bases 1 to 4456)
  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: "Cell. 2017 
            Nov 13. pii: S0092-8674(17)31255-2. doi: 
            10.1016/j.cell.2017.10.033."
COMMENT     Sequence Label: HLTV-hTRIM21
FEATURES             Location/Qualifiers
     source          1..4456
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..56
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     290..307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     392..411
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        392..410
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             442..464
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             475..492
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             769..789
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2229..2247)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2243..2290
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2387..2842
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2474..2493)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2684..2705
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2868..2972
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2973..3041,3042..3833)
                     /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(3191..3210)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4004..4456,1..136)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
       61 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      301 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
      361 tcattaatgc aggatctcga tcccgcgaaa ttaatacgac tcactatagg gagaccacaa
      421 cggtttccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcaccac
      481 caccaccacc acagcggcgc ttttgaattt aagctgccgg acattggcga aggcatccac
      541 gaaggtgaaa ttgtcaaatg gtttgtgaaa ccgggcgatg aagtgaacga agacgatgta
      601 ttgtgcgaag tgcaaaatga caaggcggtt gtcgaaattc cctccccggt caaagggaaa
      661 gtgcttgaaa tcctcgtccc ggagggaaca gtggcaacgg tcgggcaaac gctcatcacg
      721 ctcgatgcgc cgggttatga aaacatgacg accggcagcg acaccggcga aaacctgtac
      781 ttccagggtg gatctatggc ttcagcagca cgcttgacaa tgatgtggga ggaggtcaca
      841 tgccctatct gcctggaccc cttcgtggag cctgtgagca tcgagtgtgg ccacagcttc
      901 tgccaggaat gcatctctca ggttgggaaa ggtgggggca gcgtctgtcc tgtgtgccgg
      961 cagcgctttc tgctcaagaa tctccggccc aatcgacagc tagccaacat ggtgaacaac
     1021 cttaaagaaa tcagccagga ggccagagag ggcacacagg gggaacggtg tgcagtgcat
     1081 ggagagagac ttcacctgtt ctgtgagaaa gatgggaagg ccctttgctg ggtatgtgcc
     1141 cagtctcgga aacaccgtga ccacgccatg gtccctcttg aggaggctgc acaggagtac
     1201 caggagaagc tccaggtggc attaggggaa ctgagaagaa agcaggagtt ggctgagaag
     1261 ttggaagtgg aaattgcaat aaagagagca gactggaaga aaacagtgga aacacagaaa
     1321 tctaggattc acgcagagtt tgtgcagcaa aaaaacttcc tggttgaaga agaacagagg
     1381 cagctgcagg agctggagaa ggatgagagg gagcagctga gaatcctggg ggagaaagag
     1441 gccaagctgg cccagcagag ccaggcccta caggagctca tctcagagct agatcgaagg
     1501 tgccacagct cagcactgga actgctgcag gaggtgataa ttgtcctgga aaggagtgag
     1561 tcctggaacc tgaaggacct ggatattacc tctccagaac tcaggagtgt gtgccatgtg
     1621 ccagggctga agaagatgct gaggacatgt gcagtccaca tcactctgga tccagacaca
     1681 gccaatccgt ggctgatact ttcagaagat cggagacaag tgaggcttgg agacacccag
     1741 cagagcatac ctggaaatga agagagattt gatagttatc ctatggtcct gggtgcccag
     1801 cactttcact ctggaaaaca ttactgggag gtagatgtga caggaaagga ggcctgggac
     1861 ctgggtgtct gcagagactc tgtgcgcagg aaggggcact ttttgcttag ttccaagagt
     1921 ggcttctgga caatttggtt gtggaacaaa caaaaatatg aggctggcac ctacccccag
     1981 actcccctcc accttcaggt gcctccatgc caagttggga ttttcctgga ctatgaggct
     2041 ggcatggtct ccttctacaa catcactgac catggctccc tcatctactc cttctctgaa
     2101 tgtgccttta caggacctct gcggcccttc ttcagtcctg gtttcaatga tggaggaaaa
     2161 aacacagccc ctctaaccct ctgtccactg aatattggat cacaaggatc cactgactat
     2221 taagaattcc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     2281 gggttttttg ctgaaaggag gaactatatc cggatctggc gtaatagcga agaggcccgc
     2341 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     2401 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2461 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2521 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2581 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2641 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2701 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2761 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     2821 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2881 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2941 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     3001 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     3061 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     3121 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     3181 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     3241 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     3301 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     3361 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     3421 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     3481 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3541 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3601 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3661 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3721 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3781 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3841 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3901 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3961 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     4021 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     4081 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     4141 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     4201 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     4261 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     4321 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     4381 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     4441 atccggtaag cggcag
//
