LOCUS       sequence-430461-        4378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of EloB (1-104) and EloC (17-112) for production of VCB 
            complexes in conjunction with plasmid pIVM_02 (VHL 54-213).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4378)
  AUTHORS   Cardote TAF, Gadd MS, Ciulli A
  TITLE     Crystal Structure of the Cul2-Rbx1-EloBC-VHL Ubiquitin Ligase 
            Complex.
  JOURNAL   Structure. 2017 Jun 6;25(6):901-911.e3. doi: 
            10.1016/j.str.2017.04.009.
  PUBMED    28591624
REFERENCE   2  (bases 1 to 4378)
  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.str.2017.04"; journalName: "Structure"; date: "2017-06-6-
            6"; volume: "25"; issue: "6"; pages: "901-911.e3"
COMMENT     Sequence Label: pIVM_26
FEATURES             Location/Qualifiers
     source          1..4378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     567..586
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(606..683)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        729..747
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    748..772
                     /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        1202..1220
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1221..1245
                     /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)."
     CDS             1643..1687
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1725..1743)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1739..1786
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1957..2748)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2749..2840)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(join(3901..4378,1..605))
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
ORIGIN
        1 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
       61 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
      121 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
      181 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
      241 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
      301 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
      361 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
      421 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
      481 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
      541 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
      601 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
      661 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
      721 taggaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctgtaga
      781 aataattttg tttaacttta ataaggagat ataccatgga cgtgtttctc atgatccggc
      841 gccacaagac caccatcttc acggacgcca aggagtccag cacggtgttc gaactgaagc
      901 gcatcgtcga gggcatcctc aagcggcctc ctgacgagca gcggctgtac aaggatgacc
      961 aactcttgga tgatggcaag acactgggcg agtgtggctt caccagtcaa acagcacggc
     1021 cacaggcccc agccacagtg gggctggcct tccgggcaga tgacaccttc gaggccctgt
     1081 gcatcgagcc gttttccagc ccgccagagc tgcccgatgt gatgaagtag aagcttgcgg
     1141 ccgcataatg cttaagtcga acagaaagta atcgtattgt acacggccgc ataatcgaaa
     1201 ttaatacgac tcactatagg ggaattgtga gcggataaca attccccatc ttagtatatt
     1261 agttaagtat aagaaggaga tatacatatg atgtatgtca aattgatatc atctgatggc
     1321 catgaattta ttgtaaaaag agaacatgca ttaacatcag gcacgataaa agccatgttg
     1381 agtggcccag gtcagtttgc tgagaacgaa accaatgagg tcaattttag agagatacct
     1441 tcacatgtgc tatcgaaagt atgcatgtat tttacgtaca aggttcgcta cactaacagc
     1501 tccaccgaga ttcctgaatt cccaattgca cctgaaattg cactggaact gctgatggct
     1561 gcgaacttct tagattgtta gggatctcaa ttggatatcg gccggccacg cgatcgctga
     1621 cgtcggtacc ctcgagtctg gtaaagaaac cgctgctgcg aaatttgaac gccagcacat
     1681 ggactcgtct actagcgcag cttaattaac ctaggctgct gccaccgctg agcaataact
     1741 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aacctcaggc
     1801 atttgagaag cacacggtca cactgcttcc ggtagtcaat aaaccggtaa accagcaata
     1861 gacataagcg gctatttaac gaccctgccc tgaaccgacg accgggtcat cgtggccgga
     1921 tcttgcggcc cctcggcttg aacgaattgt tagacattat ttgccgacta ccttggtgat
     1981 ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg ccaagcgatc
     2041 ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat actgggccgg
     2101 caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc cggttactgc
     2161 gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag cccagtcggg
     2221 cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct gttcaggaac
     2281 cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt ctcttgcttt
     2341 tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac ctgcaagaat
     2401 gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac gccacggaat
     2461 gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc tctctccagg
     2521 ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt catcaagcct
     2581 tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc catccactgc
     2641 ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga tgacgccaac
     2701 tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcatac tcttcctttt
     2761 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     2821 tatttagaaa aataaacaaa tagctagctc actcggtcgc tacgctccgg gcgtgagact
     2881 gcggcgggcg ctgcggacac atacaaagtt acccacagat tccgtggata agcaggggac
     2941 taacatgtga ggcaaaacag cagggccgcg ccggtggcgt ttttccatag gctccgccct
     3001 cctgccagag ttcacataaa cagacgcttt tccggtgcat ctgtgggagc cgtgaggctc
     3061 aaccatgaat ctgacagtac gggcgaaacc cgacaggact taaagatccc caccgtttcc
     3121 ggcgggtcgc tccctcttgc gctctcctgt tccgaccctg ccgtttaccg gatacctgtt
     3181 ccgcctttct cccttacggg aagtgtggcg ctttctcata gctcacacac tggtatctcg
     3241 gctcggtgta ggtcgttcgc tccaagctgg gctgtaagca agaactcccc gttcagcccg
     3301 actgctgcgc cttatccggt aactgttcac ttgagtccaa cccggaaaag cacggtaaaa
     3361 cgccactggc agcagccatt ggtaactggg agttcgcaga ggatttgttt agctaaacac
     3421 gcggttgctc ttgaagtgtg cgccaaagtc cggctacact ggaaggacag atttggttgc
     3481 tgtgctctgc gaaagccagt taccacggtt aagcagttcc ccaactgact taaccttcga
     3541 tcaaaccacc tccccaggtg gttttttcga aggttaagtc agttggggaa ctgcttaacc
     3601 ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga ttacgcgcag
     3661 aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct agatttcagt
     3721 gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata agttgtaatt
     3781 ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa ggctctcaag
     3841 ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat tgcgttgcgc
     3901 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3961 cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga
     4021 gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc
     4081 cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata
     4141 acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag
     4201 cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat
     4261 cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc
     4321 actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatattta
//
