LOCUS       sequence-205567-        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-P4
FEATURES             Location/Qualifiers
     source          1..2726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    6..22
                     /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     11..33
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     30..46
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     30..46
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             42..365
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    59..115
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(116..133)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(116..132)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(125..147)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(341..360)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(500..518)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        586..690
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(691..759,760..1551)
                     /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(909..928)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1722..2310
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2211..2230
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2464..2481
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2658..2679
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2694..2699,2700..2717,2718..2724)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
       61 gcttgcatgc ctgcaggtcg actctagagg atccccgggt accgagctcg aattcactgg
      121 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
      181 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
      241 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
      301 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
      361 cataaggtct cagcttcaga ttgtcttcta agccagcccc gacacccgcc aacacccgct
      421 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgacccac
      481 gaagtgagtg gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
      541 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
      601 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
      661 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
      721 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
      781 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
      841 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
      901 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
      961 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     1021 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     1081 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     1141 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     1201 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     1261 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     1321 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     1381 gataaatctg gagccggtga gcgtggttct cgcggtatca ttgcagcact ggggccagat
     1441 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     1501 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     1561 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     1621 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     1681 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1741 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1801 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1861 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1921 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1981 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2041 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     2101 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     2161 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2221 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2281 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2341 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2401 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     2461 cgcagcgagt cagtgagcga ggaagcggat gagcgcccaa tacgcaaacc gcctctcccc
     2521 gcgcgttggc cgattcatta atcactctgt ggttcagaag acatttactt aactttaaga
     2581 aggagatata caatgggaga ccgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     2641 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     2701 tttatgcttc cggctcgtat gttgtg
//
