LOCUS       sequence-205568-        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-P5
FEATURES             Location/Qualifiers
     source          1..2726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    143..164
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(179..184,185..202,203..209)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    217..233
                     /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     222..244
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     241..257
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     241..257
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             253..576
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    270..326
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(327..344)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(327..343)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(336..358)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(552..571)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(711..729)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        797..901
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(902..970,971..1762)
                     /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(1120..1139)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1933..2521
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2422..2441
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2675..2692
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tccccgcgcg ttggccgatt cattaatcac tctgtggttc agaagacatc agattaactt
       61 taagaaggag atatacaatg ggagaccgca gctggcacga caggtttccc gactggaaag
      121 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      181 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      241 caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcaggtc gactctagag
      301 gatccccggg taccgagctc gaattcactg gccgtcgttt tacaacgtcg tgactgggaa
      361 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
      421 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
      481 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
      541 tgcactctca gtacaatctg ctctgatgcc gcataaggtc tcagctttgt gttgtcttct
      601 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
      661 ggcatccgct tacagacaag ctgtgaccca cgaagtgagt ggacgaaagg gcctcgtgat
      721 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
      781 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      841 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
      901 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
      961 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1021 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1081 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1141 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1201 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1261 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     1321 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     1381 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     1441 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     1501 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     1561 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggttc
     1621 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     1681 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     1741 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     1801 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     1861 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     1921 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     1981 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2041 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2101 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2161 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2221 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     2281 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     2341 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     2401 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     2461 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     2521 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     2581 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     2641 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     2701 tgagcgccca atacgcaaac cgcctc
//
