LOCUS       sequence-365326-        3738 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of yeast UEV-E2 complex.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3738)
  AUTHORS   Ren X, Hurley JH
  TITLE     VHS domains of ESCRT-0 cooperate in high-avidity binding to 
            polyubiquitinated cargo.
  JOURNAL   EMBO J. 2010 Mar 17;29(6):1045-54. Epub 2010 Feb 11.
  PUBMED    20150893
REFERENCE   2  (bases 1 to 3738)
  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: "EMBO J."; 
            date: "2010-03-17"; volume: "29(6)"; pages: "1045-54. Epub 2010 Feb 
            11"
COMMENT     Sequence Label: pST39-Ubc13-MMS2
FEATURES             Location/Qualifiers
     source          1..3738
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..119
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        100..118
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             150..172
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     642..659
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     643..659
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             666..688
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             708..725
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             747..767
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1241..1259)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1255..1302
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1369..1460
                     /gene="bla"
                     /label=AmpR promoter
     terminator      2325..2364
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2395..2983
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2884..2903
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3137..3154
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3169..3308
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3234..3253)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3394..3416
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3410..3601)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
       61 agatgtaggt gttggcacca tgcatcgatc ccgcgaaatt aatacgactc actataggga
      121 gaccacaacg gtttccctct agaaataatt ttgtttaact ttaagaagga gatatacata
      181 tggcatcatt acccaagaga ataatcaagg aaactgagaa attagtaagt gacccagtac
      241 ctggcattac agcagaacca catgacgaca acttacgtta cttccaagta acaatcgaag
      301 gcccagaaca gtcaccatat gaagacggta ttttcgaatt agaattatat ctgcctgacg
      361 actatccaat ggaagcaccg aaagtacgtt ttttaaccaa gatatatcat cccaatattg
      421 ataggttggg ccgtatatgt cttgatgtgc ttaaaactaa ttggtcaccc gctttacaga
      481 tcagaaccgt cctgctgtcc attcaagcac tgttggctag ccctaatcct aatgatccgc
      541 tagctaatga cgtcgctgaa gattggataa agaatgagca aggtgccaaa gctaaggctc
      601 gcgaatggac gaaattgtat gcaaagaaga aacccgagtg atgtaaaacg acggccagta
      661 ataattttgt ttaactttaa gaaggagata tacatatgtc gtactaccat caccatcacc
      721 atcacgatta cgatatccca acgaccgaaa acctgtattt tcagggcgcc atgtcaaaag
      781 taccaagaaa ttttaggttg ttagaagaat tagaaaaggg tgaaaaaggg tttggtccag
      841 aatcatgtag ttatggttta gctgatagcg acgatattac catgaccaaa tggaacggca
      901 ctattttagg gccccctcat agtaatcatg aaaacagaat ctattccctt tctatagatt
      961 gtggcccaaa ctatccagat tctcccccaa aggtcacttt catttccaag ataaatctac
     1021 catgcgtcaa tccaactacg ggggaggtcc aaactgactt ccacacctta cgtgactgga
     1081 aaagagccta caccatggaa actttattac ttgatcttag aaaggaaatg gcaaccccag
     1141 cgaacaagaa attgcgtcaa ccgaaggaag gagaaacctt ttagctcgag agatccggct
     1201 gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca
     1261 taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata
     1321 tccgactagt agcccgccta atgagcgggc ttttttttaa ttcccctatt tgtttatttt
     1381 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1441 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1501 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctcgtgaaa gtaaaagacg
     1561 cagaggacca attgggggca cgagtgggat acatagaact ggacttgaat agcggtaaaa
     1621 tccttgagag ttttcgccct gaagagcgtt ttccaatgat gagcactttc aaagttctgc
     1681 tatgtggagc agtattatcc cgtgtagatg cggggcaaga gcaactcgga cgacgaatac
     1741 actattcgca gaatgacttg gttgaatact ccccagtgac agaaaagcac cttacggacg
     1801 gaatgacggt aagagaatta tgtagtgccg ccataacgat gagtgataac actgcggcga
     1861 acttacttct gacaaccatc ggtggaccga aggaattaac cgcttttttg cacaatatgg
     1921 gagaccatgt aactcgcctt gaccgttggg aaccagaact gaatgaagcc ataccaaacg
     1981 acgagcgaga caccacaatg cctgcggcaa tggcaacaac attacgcaaa ctattaactg
     2041 gcgaactact tactctggct tcacggcaac aattaataga ctggcttgaa gcggataaag
     2101 ttgcaggacc actactgcgt tcggcacttc ctgctggctg gtttattgct gataaatctg
     2161 gggcaggaga gcgtggttca cggggtatca ttgccgcact tggaccagat ggtaagcctt
     2221 cccgtatcgt agttatctac acgacgggta gtcaggcaac tatggacgaa cgaaatagac
     2281 agattgctga aataggggct tcactgatta agcattggta aaccgataca attaaaggct
     2341 ccttttggag cctttttttt tggacggacc ggtagaaaag atcaaaggat cttcttgaga
     2401 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2461 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2521 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     2581 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2641 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2701 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2761 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2821 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2881 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2941 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3001 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3061 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3121 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
     3181 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
     3241 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     3301 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     3361 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     3421 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     3481 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     3541 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     3601 ctgatgcctc cgtgtaaggg ggatttctgt tcatggggta atgataccga tgaaacgaga
     3661 gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga
     3721 gggtaaacaa ctggcggt
//
