LOCUS       sequence-205598-        3320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cloning Module.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3320)
  AUTHORS   Kowarschik K, Hoehenwarter W, Marillonnet S, Trujillo M
  TITLE     UbiGate: a synthetic biology toolbox to analyse ubiquitination.
  JOURNAL   New Phytol. 2018 Mar;217(4):1749-1763. doi: 10.1111/nph.14900. Epub 
            2017 Nov 30.
  PUBMED    29194629
REFERENCE   2  (bases 1 to 3320)
  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: "10.1111/nph.14900";
            journalName: "New Phytol"; date: "2018-03"; volume: "217"; issue: 
            "4"; pages: "1749-1763"
COMMENT     Sequence Label: pUG41766
FEATURES             Location/Qualifiers
     source          1..3320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     104..121
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    238..259
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(274..279,280..297,298..304)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    312..328
                     /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)."
     primer_bind     317..339
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     336..352
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     336..352
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(904..921)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(904..920)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(913..935)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1129..1148)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1248..1270
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1308..1326)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1394..1498
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1720..2511
                     /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"
     rep_origin      2682..3270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3171..3190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
       61 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      121 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      181 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      241 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
      301 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
      361 cgccaagctt gcatgcctgc aggtcgactc tagaggatcc cctttgaaga caattacgag
      421 gatgcacatg tgaccgaggg attgtcttcg tggaaacttg ggacttcaga ttccacccct
      481 tgcaagcgtg gaaagccttc cgaccaaggg aagtgtcgga tgtagaggaa atggaaagtt
      541 tgttctcaga tggggacttg cttgattgct tcacaagaat gccagcttat gcggtaaacg
      601 cagaggaaga tttagctgca atcaggaaaa cgcccgagat ggatgtcggt caagaagtta
      661 aagaacctgc aggagacaga aatcaatact caaaccctgc agaaactttc ctcaacaagc
      721 tccacaggaa acacagtagg gaggtgaaac accaggccgc aaagaaagct aaacgcctag
      781 ctgaaatcca ggagtcaatg agagctgaag gtgatgccga accaaatgaa ataagcgggg
      841 gcatgggggc aatacccagc aacgccgaac ttcctgggcc caaagggtac cgagctcgaa
      901 ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa
      961 tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga
     1021 tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct
     1081 ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc
     1141 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     1201 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accatctccg ggagctgcat
     1261 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagt cgaaagggcc tcgtgatacg
     1321 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     1381 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1441 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     1501 gcgctcacgc aactggtcca gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc
     1561 ggttttcatg gcttgttatg actgtttttt tttggggtac agtctatgcc tcgggcatcc
     1621 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
     1681 acgcagcagg gcagtcgccc taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc
     1741 gaagtatcga ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg
     1801 ttgctggccg tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat
     1861 attgatttgc tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc
     1921 aacgaccttt tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa
     1981 gtcaccattg ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg
     2041 caatttggag aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc
     2101 gacattgatc tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt
     2161 ccagcggcgg aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat
     2221 gaaaccttaa cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg
     2281 cttacgttgt cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc
     2341 gctgccgact gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct
     2401 agacaggctt atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa
     2461 gaatttgtcc actacgtgaa aggcgagatc accaaggtag tcggcaaata actgtcagac
     2521 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     2581 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     2641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2761 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2821 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2881 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2941 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3001 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3061 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3241 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3301 ctggcctttt gctggccttt
//
