LOCUS       sequence-205467-        2726 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 cloning vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2726)
  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 2726)
  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-P3
FEATURES             Location/Qualifiers
     source          1..2726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    181..202
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(217..222,223..240,241..247)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    255..271
                     /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     260..282
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     279..295
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     279..295
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             291..614
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    308..364
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(365..382)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(365..381)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(374..396)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(590..609)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(749..767)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        835..939
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(940..1008,1009..1800)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1158..1177)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1971..2559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2460..2479
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(2713..2726,1..4)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgaggaagcg gatgagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
       61 ttaatcactc tgtggttcag aagacatact attaacttta agaaggagat atacaatggg
      121 agaccgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
      181 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
      241 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      301 ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta ccgagctcga
      361 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
      421 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
      481 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
      541 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
      601 ctgatgccgc ataaggtctc agcttttact tgtcttctaa gccagccccg acacccgcca
      661 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
      721 gtgacccacg aagtgagtgg acgaaagggc ctcgtgatac gcctattttt ataggttaat
      781 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
      841 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
      901 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
      961 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1021 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1081 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1141 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     1201 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1261 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     1321 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     1381 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     1441 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     1501 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     1561 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     1621 tttattgctg ataaatctgg agccggtgag cgtggttctc gcggtatcat tgcagcactg
     1681 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     1741 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     1801 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     1861 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     1921 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     1981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2041 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2101 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2161 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2221 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2281 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2341 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2401 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2461 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2521 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2581 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     2641 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     2701 acgaccgagc gcagcgagtc agtgag
//
