LOCUS       sequence-205601-        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: pUG41822
FEATURES             Location/Qualifiers
     source          1..3320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..73
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    190..211
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(226..231,232..249,250..256)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    264..280
                     /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     269..291
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     288..304
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     288..304
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(856..873)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(856..872)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(865..887)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1081..1100)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1200..1222
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1260..1278)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1346..1450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1672..2463
                     /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      2634..3222
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3123..3142
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
       61 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
      121 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
      181 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
      241 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
      301 tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac tctagaggat cccctttggg
      361 cccaggaagt tcggcgttgc tgggtattgc ccccatgccc ccgcttattt catttggttc
      421 ggcatcacct tcagctctca ttgactcctg gatttcagct aggcgtttag ctttctttgc
      481 ggcctggtgt ttcacctccc tactgtgttt cctgtggagc ttgttgagga aagtttctgc
      541 agggtttgag tattgatttc tgtctcctgc aggttcttta acttcttgac cgacatccat
      601 ctcgggcgtt ttcctgattg cagctaaatc ttcctctgcg tttaccgcat aagctggcat
      661 tcttgtgaag caatcaagca agtccccatc tgagaacaaa ctttccattt cctctacatc
      721 cgacacttcc cttggtcgga aggctttcca cgcttgcaag gggtggaatc tgaagtccca
      781 agtttccacg aagacaatcc ctcggtcaca tgtgcatcct cgctcttgtc ttcaaagggt
      841 accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
      901 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
      961 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat
     1021 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     1081 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     1141 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccatctc
     1201 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gtcgaaaggg
     1261 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1321 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1381 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1441 aaggaagagt atgcgctcac gcaactggtc cagaaccttg accgaacgca gcggtggtaa
     1501 cggcgcagtg gcggttttca tggcttgtta tgactgtttt tttttggggt acagtctatg
     1561 cctcgggcat ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc
     1621 agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa agttaaacat catgagggaa
     1681 gcggtgatcg ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat
     1741 ctcgaaccga cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag
     1801 ccacacagtg atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg
     1861 cgagctttga tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc
     1921 cgcgctgtag aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct
     1981 aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag
     2041 ccagccacga tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt
     2101 gccttggtag gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt
     2161 gaggcgctaa atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag
     2221 cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg
     2281 ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc
     2341 atacttgaag ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca
     2401 gatcagttgg aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa
     2461 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2521 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2581 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2641 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2701 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2761 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2821 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2881 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2941 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3001 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3061 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3121 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3181 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3241 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     3301 cctgattctg tggataaccg
//
