LOCUS       sequence-205597-        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: pUG41744
FEATURES             Location/Qualifiers
     source          1..3320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(335..439)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     507..525
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(563..585)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     685..704
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     898..920
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     912..929
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     913..929
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1481..1497)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1481..1497)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1494..1516)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1505..1521
                     /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(1529..1535,1536..1553,1554..1559))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1574..1595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1712..1729)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1883..2471)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1963..1982)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2642..3320,1..113))
                     /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 actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg ctcgatgacg
       61 ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct catgatgttt
      121 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca taacatcaaa
      181 catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga ctgtacccca
      241 aaaaaaaaca gtcataacaa gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt
      301 cggtcaaggt tctggaccag ttgcgtgagc gcatactctt cctttttcaa tattattgaa
      361 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
      421 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
      481 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgactcgcgc
      541 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagatg gtcacagctt
      601 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
      661 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
      721 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
      781 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
      841 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
      901 agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtaccctt tgaagacaaa
      961 ctagaggatg cacatgtgac cgagggattg tcttcgtgga aacttgggac ttcagattcc
     1021 accccttgca agcgtggaaa gccttccgac caagggaagt gtcggatgta gaggaaatgg
     1081 aaagtttgtt ctcagatggg gacttgcttg attgcttcac aagaatgcca gcttatgcgg
     1141 taaacgcaga ggaagattta gctgcaatca ggaaaacgcc cgagatggat gtcggtcaag
     1201 aagttaaaga acctgcagga gacagaaatc aatactcaaa ccctgcagaa actttcctca
     1261 acaagctcca caggaaacac agtagggagg tgaaacacca ggccgcaaag aaagctaaac
     1321 gcctagctga aatccaggag tcaatgagag ctgaaggtga tgccgaacca aatgaaataa
     1381 gcgggggcat gggggcaata cccagcaacg ccgaacttcc tgggcccaaa ggggatcctc
     1441 tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     1501 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     1561 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     1621 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1681 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1741 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1801 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1861 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1921 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1981 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     2041 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     2101 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     2161 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     2221 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     2281 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2341 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2401 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2461 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2521 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2581 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2641 gttatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact
     2701 gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa
     2761 gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct
     2821 tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat
     2881 ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg
     2941 cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca
     3001 aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg
     3061 ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc
     3121 gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag
     3181 cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag
     3241 ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt
     3301 gtggcttcag gccgccatcc
//
