LOCUS       sequence-205600-        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: pUG41800
FEATURES             Location/Qualifiers
     source          1..3320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(334..438)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     506..524
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(562..584)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     684..703
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     897..919
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     911..928
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     912..928
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1480..1496)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1480..1496)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1493..1515)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1504..1520
                     /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        complement(join(1528..1534,1535..1552,1553..1558))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1573..1594
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1711..1728)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1882..2470)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1962..1981)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2641..3320,1..112))
                     /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"
ORIGIN
        1 ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc
       61 caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc atgatgttta
      121 actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctccat aacatcaaac
      181 atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtaccccaa
      241 aaaaaaacag tcataacaag ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc
      301 ggtcaaggtt ctggaccagt tgcgtgagcg catactcttc ctttttcaat attattgaag
      361 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
      421 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
      481 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gactcgcgcg
      541 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagatgg tcacagcttg
      601 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
      661 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
      721 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
      781 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
      841 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
      901 gtcacgacgt tgtaaaacga cggccagtga attcgagctc ggtacccttt gggcccagga
      961 agttcggcgt tgctgggtat tgcccccatg cccccgctta tttcatttgg ttcggcatca
     1021 ccttcagctc tcattgactc ctggatttca gctaggcgtt tagctttctt tgcggcctgg
     1081 tgtttcacct ccctactgtg tttcctgtgg agcttgttga ggaaagtttc tgcagggttt
     1141 gagtattgat ttctgtctcc tgcaggttct ttaacttctt gaccgacatc catctcgggc
     1201 gttttcctga ttgcagctaa atcttcctct gcgtttaccg cataagctgg cattcttgtg
     1261 aagcaatcaa gcaagtcccc atctgagaac aaactttcca tttcctctac atccgacact
     1321 tcccttggtc ggaaggcttt ccacgcttgc aaggggtgga atctgaagtc ccaagtttcc
     1381 acgaagacaa tccctcggtc acatgtgcat cctccacatt gtcttcaaag gggatcctct
     1441 agagtcgacc tgcaggcatg caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     1501 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     1561 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     1621 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     1681 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     1741 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     1801 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     1861 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     1921 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1981 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2041 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2101 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2161 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     2221 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     2281 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     2341 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     2401 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     2461 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     2521 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     2581 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     2641 ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg
     2701 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag
     2761 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt
     2821 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt
     2881 tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc
     2941 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa
     3001 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc
     3061 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg
     3121 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc
     3181 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc
     3241 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg
     3301 tggcttcagg ccgccatcca
//
