LOCUS       sequence-205596-        3319 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 3319)
  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 3319)
  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: pUG41722
FEATURES             Location/Qualifiers
     source          1..3319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    110..131
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(146..151,152..169,170..176)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    184..200
                     /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     189..211
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     208..224
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     208..224
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(775..792)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(775..791)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(784..806)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1000..1019)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1119..1141
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1179..1197)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1265..1369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1591..2382
                     /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      2553..3141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3042..3061
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3295..3312
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
       61 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      121 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      181 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      241 ttgcatgcct gcaggtcgac tctagaggat ccccttgaag acaagcaaga ggatgcacat
      301 gtgaccgagg gattgtcttc gtggaaactt gggacttcag attccacccc ttgcaagcgt
      361 ggaaagcctt ccgaccaagg gaagtgtcgg atgtagagga aatggaaagt ttgttctcag
      421 atggggactt gcttgattgc ttcacaagaa tgccagctta tgcggtaaac gcagaggaag
      481 atttagctgc aatcaggaaa acgcccgaga tggatgtcgg tcaagaagtt aaagaacctg
      541 caggagacag aaatcaatac tcaaaccctg cagaaacttt cctcaacaag ctccacagga
      601 aacacagtag ggaggtgaaa caccaggccg caaagaaagc taaacgccta gctgaaatcc
      661 aggagtcaat gagagctgaa ggtgatgccg aaccaaatga aataagcggg ggcatggggg
      721 caatacccag caacgccgaa cttcctgggc ccaaagggta ccgagctcga attcactggc
      781 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
      841 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
      901 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
      961 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     1021 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     1081 gctcccggca tccgcttaca gacaagctgt gaccatctcc gggagctgca tgtgtcagag
     1141 gttttcaccg tcatcaccga aacgcgcgag tcgaaagggc ctcgtgatac gcctattttt
     1201 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1261 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1321 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgcgctcacg
     1381 caactggtcc agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat
     1441 ggcttgttat gactgttttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa
     1501 gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag
     1561 ggcagtcgcc ctaaaacaaa gttaaacatc atgagggaag cggtgatcgc cgaagtatcg
     1621 actcaactat cagaggtagt tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc
     1681 gtacatttgt acggctccgc agtggatggc ggcctgaagc cacacagtga tattgatttg
     1741 ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc gagctttgat caacgacctt
     1801 ttggaaactt cggcttcccc tggagagagc gagattctcc gcgctgtaga agtcaccatt
     1861 gttgtgcacg acgacatcat tccgtggcgt tatccagcta agcgcgaact gcaatttgga
     1921 gaatggcagc gcaatgacat tcttgcaggt atcttcgagc cagccacgat cgacattgat
     1981 ctggctatct tgctgacaaa agcaagagaa catagcgttg ccttggtagg tccagcggcg
     2041 gaggaactct ttgatccggt tcctgaacag gatctatttg aggcgctaaa tgaaacctta
     2101 acgctatgga actcgccgcc cgactgggct ggcgatgagc gaaatgtagt gcttacgttg
     2161 tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac
     2221 tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca tacttgaagc tagacaggct
     2281 tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc
     2341 cactacgtga aaggcgagat caccaaggta gtcggcaaat aactgtcaga ccaagtttac
     2401 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2461 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2521 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2581 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2641 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2701 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2761 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2821 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2881 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     2941 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3001 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3061 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3121 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3181 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3241 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3301 tcagtgagcg aggaagcgg
//
