LOCUS       sequence-198489-        3318 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 3318)
  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 3318)
  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: pUG41744
FEATURES             Location/Qualifiers
     source          1..3318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(51..67)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(51..67)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(64..86)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    75..91
                     /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(99..105,106..123,124..129))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    144..165
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(282..299)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(453..1041)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(533..552)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1212..2003)
                     /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(2223..2327)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2395..2413
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2451..2473)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2573..2592
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2786..2808
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2800..2817
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2801..2817
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg
       61 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
      121 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
      181 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
      241 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
      301 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      361 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      421 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
      481 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      541 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      601 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      661 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      721 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      781 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      841 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
      901 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      961 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1021 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1081 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     1141 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1201 tggtctgaca gttatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat
     1261 tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt
     1321 ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca
     1381 gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta
     1441 agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt
     1501 tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct
     1561 ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg
     1621 tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat
     1681 tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg
     1741 acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg
     1801 ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca
     1861 atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc
     1921 aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact
     1981 tcggcgatca ccgcttccct catgatgttt aactttgttt tagggcgact gccctgctgc
     2041 gtaacatcgt tgctgctcca taacatcaaa catcgaccca cggcgtaacg cgcttgctgc
     2101 ttggatgccc gaggcataga ctgtacccca aaaaaacagt cataacaagc catgaaaacc
     2161 gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc
     2221 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     2281 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     2341 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     2401 cgtatcacga ggccctttcg actcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     2461 atgcagctcc cggagatggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     2521 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     2581 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     2641 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     2701 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     2761 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     2821 ttcgagctcg gtaccctttg aagacaaact agaggatgca catgtgaccg agggattgtc
     2881 ttcgtggaaa cttgggactt cagattccac cccttgcaag cgtggaaagc cttccgacca
     2941 agggaagtgt cggatgtaga ggaaatggaa agtttgttct cagatgggga cttgcttgat
     3001 tgcttcacaa gaatgccagc ttatgcggta aacgcagagg aagatttagc tgcaatcagg
     3061 aaaacgcccg agatggatgt cggtcaagaa gttaaagaac ctgcaggaga cagaaatcaa
     3121 tactcaaacc ctgcagaaac tttcctcaac aagctccaca ggaaacacag tagggaggtg
     3181 aaacaccagg ccgcaaagaa agctaaacgc ctagctgaaa tccaggagtc aatgagagct
     3241 gaaggtgatg ccgaaccaaa tgaaataagc gggggcatgg gggcaatacc cagcaacgcc
     3301 gaacttcctg ggcccaaa
//
