LOCUS       Exported                3468 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Cloning Module.
ACCESSION   .
VERSION     .
KEYWORDS    pUG-225
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3468)
  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 3468)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3468
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     103..120
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             199..1299
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1273..1296
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1348..1359
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1455..1473)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1541..1645
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1867..2658
                     /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      2829..3417
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3318..3337
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
       61 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
      121 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
      181 tcactctgtg gtctcaccat gaaaatcgaa gaaggtaaac tggtaatctg gattaacggc
      241 gataaaggct ataacggact cgctgaagtc ggtaagaaat tcgagaaaga taccggaatt
      301 aaagtcaccg ttgagcatcc ggataaactg gaagagaaat tcccacaggt tgcggcaact
      361 ggcgatggcc ctgacattat cttctgggca cacgaccgct ttggtggcta cgctcaatct
      421 ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg acaagctgta tccgtttacc
      481 tgggatgccg tacgttacaa cggcaagctg attgcttacc cgatcgctgt tgaagcgtta
      541 tcgctgattt ataacaaaga tctgctgccg aacccgccaa aaacctggga agagatcccg
      601 gcgctggata aagaactgaa agcgaaaggt aagagcgcgc tgatgttcaa cctgcaagaa
      661 ccgtacttca cctggccgct gattgctgct gacgggggtt atgcgttcaa gtatgaaaac
      721 ggcaagtacg acattaaaga cgtgggcgtg gataacgctg gcgcgaaagc gggtctgacc
      781 ttcctggttg acctgattaa aaacaaacac atgaatgcag acaccgatta ctccatcgca
      841 gaagctgcct ttaataaagg cgaaacagcg atgaccatca acggcccgtg ggcatggtcc
      901 aacatcgaca ccagcaaagt gaattatggt gtaacggtac tgccgacctt caagggtcaa
      961 ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta acgccgccag tccgaacaaa
     1021 gagctggcaa aagagttcct cgaaaactat ctgctgactg atgaaggtct ggaagcggtt
     1081 aataaagaca aaccgctggg tgccgtagcg ctgaagtctt acgaggaaga gttggcgaaa
     1141 gatccacgta ttgccgccac tatggaaaac gcccagaaag gtgaaatcat gccgaacatc
     1201 ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg tgatcaacgc cgccagcggt
     1261 cgtcagactg tcgatgaagc cctgaaagac gcgcagacta attcgagctc gaacaacaac
     1321 aacaataaca ataacaacaa cctcgggatc gagggaagga tttcagaatt cggatcctct
     1381 agagtcgacc tgcaggcaag cttggcactg gccgtaatgt gagaccacga agtggctctt
     1441 cagtggacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     1501 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1561 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1621 tcaataatat tgaaaaagga agagtatgcg ctcacgcaac tggtccagaa ccttgaccga
     1681 acgcagcggt ggtaacggcg cagtggcggt tttcatggct tgttatgact gttttttttt
     1741 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
     1801 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
     1861 aacatcatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
     1921 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
     1981 gatggcggcc tgaagccaca cagcgatatt gatttgctgg ttacggtgac cgtaaggctt
     2041 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
     2101 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
     2161 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
     2221 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
     2281 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
     2341 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
     2401 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
     2461 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
     2521 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
     2581 ttggcctcgc gcgcagatca gttggaagaa tttgtccatt acgtaaaagg cgagatcacc
     2641 aaggtagtcg gcaaataact gtcagaccaa gtttactcat atatacttta gattgattta
     2701 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     2761 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     2821 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     2881 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     2941 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     3001 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3061 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3121 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3181 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     3241 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3301 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3361 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     3421 gccagcaacg cggccttttt acggttcctg gccttttgct ggcctttt
//
