LOCUS       sequence-207042-        2847 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 cloning vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2847)
  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 2847)
  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: pUG-1287
FEATURES             Location/Qualifiers
     source          1..2847
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     50..67
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    211..232
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(247..252,253..270,271..277)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    285..301
                     /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     290..312
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     309..325
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     309..325
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             321..644
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    338..394
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(395..412)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(395..411)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(404..426)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(620..639)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(781..799)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        867..971
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1193..1984
                     /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      2155..2743
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2644..2663
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
       61 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
      121 tcattaatca ctctgtggtc tcaaatgttg tcttcgcagc tggcacgaca ggtttcccga
      181 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
      241 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
      301 atttcacaca ggaaacagct atgaccatga ttacgccaag cttgcatgcc tgcaggtcga
      361 ctctagagga tccccgggta ccgagctcga attcactggc cgtcgtttta caacgtcgtg
      421 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
      481 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
      541 atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
      601 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
      661 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
      721 gacaagctgt gacgaagaca attcgtgaga ccacgaagtg gctcttcagt ggacgaaagg
      781 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
      841 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
      901 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      961 aaaggaagag tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta
     1021 acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttttgggg tacagtctat
     1081 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     1141 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca tcatgaggga
     1201 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     1261 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     1321 gccacacagc gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     1381 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     1441 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     1501 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     1561 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     1621 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     1681 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     1741 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     1801 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     1861 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     1921 agatcagttg gaagaatttg tccattacgt aaaaggcgag atcaccaagg tagtcggcaa
     1981 ataactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2041 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2101 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2161 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2221 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2281 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     2341 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2401 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2461 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2521 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2581 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2641 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     2701 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     2761 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     2821 ccctgattct gtggataacc gtattac
//
