LOCUS       sequence-210889-        3891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  E. coli expression and purification of 6xTR-TUBE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3891)
  AUTHORS   Tsuchiya H, Burana D, Ohtake F, Arai N, Kaiho A, Komada M, Tanaka K,
            Saeki Y
  TITLE     Ub-ProT reveals global length and composition of protein 
            ubiquitylation in cells.
  JOURNAL   Nat Commun. 2018 Feb 6;9(1):524. doi: 10.1038/s41467-018-02869-x.
  PUBMED    29410401
REFERENCE   2  (bases 1 to 3891)
  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: "Nat 
            Commun."; date: "2018-02-6"; volume: "9(1):524. doi"; pages: " 
            10.1038/s41467-018-02869-x"
COMMENT     Sequence Label: pRSET-6xTR-TUBE
FEATURES             Location/Qualifiers
     source          1..3891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             70..92
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             112..129
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             133..165
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     terminator      1306..1353
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1450..1905
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1537..1556)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1747..1768
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1931..2035
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2036..2104,2105..2896)
                     /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(2254..2273)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3067..3655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3556..3575
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3809..3826
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaaataatt ttgtttaact ttaagaagga gatatacata tgcggggttg tcatcatcat
      121 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtgatat cggcggcggt
      181 ggttcgggag gtggggtaaa tccgcagctg cagaatcctg aggtcgcgtt tcaacagcag
      241 ttggagcaat tgagcgcaat ggggttctta aatgcggaag cgaaccttca ggccctcatt
      301 gccaccggtg gggatattaa tgcggccatt gaagccctgc tgggttcgca gccatccggc
      361 ggcggcgggt ctggtggagg ggtgaaccct cagttacaga atccggaagt cgcgtttcag
      421 caacaattag agcagttaag cgccatgggc tttcttaacg cggaagcgaa cttgcaggcc
      481 ctgatcgcga ctggtggaga tattaatgca gccatcgagg cacttctggg gagccagccc
      541 agtggcggcg gcgggagcgg tggaggtgtg aacccgcagt tgcaaaaccc ggaagtggcg
      601 ttccaacagc agctggaaca actgtcggcc atgggttttc tgaacgcaga agctaacctc
      661 caagcgctga ttgctaccgg tggcgatatt aacgctgcga tcgaagcact gctcggctca
      721 cagccaagtg gcggtggcgg cagcggtggc ggcgttaatc cgcaactgca aaaccccgaa
      781 gttgcgtttc agcagcaact ggagcagctg tccgcaatgg gcttcctgaa tgccgaagca
      841 aatctgcagg ctctcattgc tacgggtggc gacatcaacg ctgccatcga agcgctgctg
      901 ggctctcagc cgtcaggagg tggtggatct ggtggaggcg tgaatcccca gcttcagaac
      961 ccagaagtag cctttcagca gcaactggag caactgtccg ctatggggtt tctgaacgct
     1021 gaagcgaact tgcaagccct gatcgcaacc ggtggcgaca tcaatgcagc catcgaagcc
     1081 ttgttaggca gccaaccttc aggaggtggt ggctctggcg gtggcgtcaa tccgcagctg
     1141 cagaatccgg aagttgcgtt ccaacagcag ctggaacagc tcagcgcaat gggcttcctg
     1201 aacgcggaag ctaaccttca ggcgctcatt gcaacgggtg gcgatattaa cgcggccatt
     1261 gaggcgttac tggggtcgca accgagtggc ggtgggggat cctaactagc ataacccctt
     1321 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatct
     1381 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
     1441 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1501 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1561 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1621 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1681 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1741 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1801 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1861 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg
     1921 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     1981 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2041 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2101 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2161 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2221 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2281 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2341 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2401 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2461 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2521 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2581 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2641 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2701 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     2761 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     2821 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     2881 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     2941 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3001 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3061 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3121 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3181 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     3241 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3301 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3361 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3421 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3481 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3541 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3601 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3661 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3721 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     3781 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     3841 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca g
//
