LOCUS       sequence-409504-        5117 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of UBE2G2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5117)
  AUTHORS   Das R, Mariano J, Tsai YC, Kalathur RC, Kostova Z, Li J, Tarasov SG,
            McFeeters RL, Altieri AS, Ji X, Byrd RA, Weissman AM
  TITLE     Allosteric activation of E2-RING finger-mediated ubiquitylation by a
            structurally defined specific E2-binding region of gp78.
  JOURNAL   Mol Cell. 2009 Jun 26. 34(6):674-85.
  PUBMED    19560420
REFERENCE   2  (bases 1 to 5117)
  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: "Mol Cell. 
            2009 Jun 26. 34(6):674-85."
COMMENT     Sequence Label: pET3a-UBE2G2
FEATURES             Location/Qualifiers
     source          1..5117
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             473..490
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(544..562)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      558..605
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(971..976,977..993,994..999))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1026..1044)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1112..1216
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1217..1285,1286..2077)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1435..1454)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2248..2836
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2737..2756
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2990..3007
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3022..3161
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3087..3106)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3247..3269
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3263..3454)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     4912..4931
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5015..5034
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5015..5033
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             5065..5087
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 gctgatggcc gagtataagc aattaaccct gaatcctcca gaaggaatcg tggcaggccc
       61 catgaatgaa gagaattttt ttgaatggga ggcattgatc atgggccctg aagacacttg
      121 ttttgagttt ggtgtttttc ccgccatcct gagtttccca cttgactacc ccttgagccc
      181 tccgaagatg agattcacct gtgagatgtt ccatcccaac atctatcctg atggcagagt
      241 ctgcatctcc atcctgcatg ctccaggtga tgatcccatg ggttatgaga gcagtgccga
      301 gcggtggagc ccagtgcaga gtgtggagaa gatcctgctt tctgttgtga gcatgctggc
      361 agagcccaac gatgagagtg gagcaaacgt agatgcttcc aagatgtggc gggatgaccg
      421 ggagcagttc tacaagatcg ccaagcagat tgtccagaag tctctggggc tccatcacca
      481 tcaccatcac tgaggatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
      541 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
      601 ttttgctgaa aggaggaact atatccggat atccacagga cgggtgtggt cgccatgatc
      661 gcgtagtcga tagtggctcc aagtagcgaa gcgagcagga ctgggcggcg gccaaagcgg
      721 tcggacagtg ctccgagaac gggtgcgcat agaaattgca tcaacgcata tagcgctagc
      781 agcacgccat agtgactggc gatgctgtcg gaatggacga tatcccgcaa gaggcccggc
      841 agtaccggca taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg
      901 atgagcgcat tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat
      961 aaactaccgc attaaagctt atcgatgata agctgtcaaa catgagaatt cttgaagacg
     1021 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     1081 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     1141 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     1201 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     1261 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     1321 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     1381 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     1441 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     1501 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     1561 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     1621 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     1681 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     1741 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     1801 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     1861 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     1921 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     1981 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     2041 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     2101 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2161 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2221 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     2281 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     2341 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     2401 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     2461 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     2521 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     2581 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     2641 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     2701 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     2761 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     2821 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     2881 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     2941 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     3001 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     3061 ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3121 tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac
     3181 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     3241 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc
     3301 agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat
     3361 ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg
     3421 ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc
     3481 tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg
     3541 atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc
     3601 ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag
     3661 gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg
     3721 gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg
     3781 ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg
     3841 attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga
     3901 gcacgatcat gcgcacccgt ggccaggacc caacgctgcc cgagatgcgc cgcgtgcggc
     3961 tgctggagat ggcggacgcg atggatatgt tctgccaagg gttggtttgc gcattcacag
     4021 ttctccgcaa gaattgattg gctccaattc ttggagtggt gaatccgtta gcgaggtgcc
     4081 gccggcttcc attcaggtcg aggtggcccg gctccatgca ccgcgacgca acgcggggag
     4141 gcagacaagg tatagggcgg cgcctacaat ccatgccaac ccgttccatg tgctcgccga
     4201 ggcggcataa atcgccgtga cgatcagcgg tccaatgatc gaagttaggc tggtaagagc
     4261 cgcgagcgat ccttgaagct gtccctgatg gtcgtcatct acctgcctgg acagcatggc
     4321 ctgcaacgcg ggcatcccga tgccgccgga agcgagaaga atcataatgg ggaaggccat
     4381 ccagcctcgc gtcgcgaacg ccagcaagac gtagcccagc gcgtcggccg ccatgccggc
     4441 gataatggcc tgcttctcgc cgaaacgttt ggtggcggga ccagtgacga aggcttgagc
     4501 gagggcgtgc aagattccga ataccgcaag cgacaggccg atcatcgtcg cgctccagcg
     4561 aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta cgagttgcat
     4621 gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc accggaagga
     4681 gctgactggg ttgaaggctc tcaagggcat cggtcgacgc tctcccttat gcgactcctg
     4741 cattaggaag cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg
     4801 tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac
     4861 gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc
     4921 ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc
     4981 ggcgtagagg atcgagatct cgatcccgcg aaattaatac gactcactat agggagacca
     5041 caacggtttc cctctagaaa taattttgtt taactttaag aaggagatat acatatggcg
     5101 gggacggcgt tgaagag
//
