LOCUS       sequence-459913-        4307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression AtUBC8-S in in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4307)
  AUTHORS   Pu ZX, Wang JL, Li YY, Liang LY, Tan YT, Wang ZH, Li BL, Guo GQ, 
            Wang L, Wu L
  TITLE     A Bacterial Platform for Studying Ubiquitination Cascades Anchored 
            by SCF-Type E3 Ubiquitin Ligases.
  JOURNAL   Biomolecules. 2024 Sep 25;14(10):1209. doi: 10.3390/biom14101209.
  PUBMED    39456142
REFERENCE   2  (bases 1 to 4307)
  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: 
            "Biomolecules."; date: "2024-09-25"; volume: "14(10):1209. doi"; 
            pages: " 10.3390/biom14101209"
COMMENT     Sequence Label: pCDFDuet-AtUBC8-S
FEATURES             Location/Qualifiers
     source          1..4307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        46..64
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    65..89
                     /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             145..162
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             637..681
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     promoter        802..820
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    821..845
                     /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             954..998
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1036..1054)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1050..1097
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1268..2059)
                     /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(2060..2151)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2199..2937)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(3147..4229)
                     /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"
     primer_bind     4191..4210
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4230..4307)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 cgggatctcg acgctctccc ttatgcgact cctgcattag gaaattaata cgactcacta
       61 taggggaatt gtgagcggat aacaattccc ctgtagaaat aattttgttt aactttaata
      121 aggagatata ccatgggcag cagccatcac catcatcacc acagccagga tccgaattcc
      181 atggcttcga aacggatctt gaaggagctg aaggatctcc agaaagaccc tccaacctcc
      241 tgcagtgcag gtccagttgc tgaagacatg tttcattggc aagctacaat tatgggtcct
      301 gcagagagtc cgtattcagg cggtgtgttt ctcgttacca ttcacttccc tccggactat
      361 ccattcaaac caccaaaggt tgcatttagg acgaaggtgt ttcaccctaa tatcaacagc
      421 aacggaagca tttgccttga cattttgaaa gaacaatgga gccctgccct caccatttcc
      481 aaggtgttgc tctcgatatg ttcgctgtta acagatccaa atccagatga ccctttggta
      541 ccagagattg cacacatgta caaaaccgac agagccaaat acgaggctac tgcaagaaac
      601 tggactcaga agtatgccat gggcctcgag tctggtaaag aaaccgctgc tgcgaaattt
      661 gaacgccagc acatggactc gtctactagc gcagcttaag aattcgagct cggcgcgcct
      721 gcaggtcgac aagcttgcgg ccgcataatg cttaagtcga acagaaagta atcgtattgt
      781 acacggccgc ataatcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
      841 attccccatc ttagtatatt agttaagtat aagaaggaga tatacatatg gcagatctca
      901 attggatatc ggccggccac gcgatcgctg acgtcggtac cctcgagtct ggtaaagaaa
      961 ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc tactagcgca gcttaattaa
     1021 cctaggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1081 tcttgagggg ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc
     1141 cggtagtcaa taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc
     1201 ctgaaccgac gaccgggtca tcgtggccgg atcttgcggc ccctcggctt gaacgaattg
     1261 ttagacatta tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt
     1321 ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag
     1381 cttcaagtat gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga
     1441 catccttcgg cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca
     1501 ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat
     1561 ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac
     1621 ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga
     1681 tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca
     1741 gttcgcgctt agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt
     1801 ctacagcgcg gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga
     1861 tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat
     1921 cactgtgtgg cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg
     1981 tcggttcgag atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg
     2041 cgatcaccgc ttccctcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     2101 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa atagctagct
     2161 cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt
     2221 tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc
     2281 gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt
     2341 ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac
     2401 ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg
     2461 ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc
     2521 gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg
     2581 ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca
     2641 cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg
     2701 gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt
     2761 ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt
     2821 taagcagttc cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg
     2881 tttacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     2941 tctactgaac cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
     3001 agccccatac gatataagtt gtaattctca tgttagtcat gccccgcgcc caccggaagg
     3061 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga
     3121 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     3181 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
     3241 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
     3301 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
     3361 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
     3421 accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
     3481 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
     3541 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
     3601 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
     3661 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
     3721 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
     3781 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
     3841 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
     3901 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
     3961 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
     4021 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
     4081 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
     4141 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
     4201 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
     4261 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtc
//
