LOCUS       sequence-205580-        2725 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 2725)
  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 2725)
  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-TP3
FEATURES             Location/Qualifiers
     source          1..2725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    148..169
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(184..189,190..207,208..214)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    222..238
                     /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     227..249
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     246..262
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     246..262
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             258..581
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    275..331
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(332..349)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(332..348)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(341..363)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(557..576)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(716..734)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        802..906
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(907..975,976..1767)
                     /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(1125..1144)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1938..2526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2427..2446
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2680..2697
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcctctccc cgcgcgttgg ccgattcatt aatcactctg tggttcagaa gacatactat
       61 taactttaag aaggagatat accatggaga ccgcagctgg cacgacaggt ttcccgactg
      121 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      181 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      241 tcacacagga aacagctatg accatgatta cgccaagctt gcatgcctgc aggtcgactc
      301 tagaggatcc ccgggtaccg agctcgaatt cactggccgt cgttttacaa cgtcgtgact
      361 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
      421 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctgaatg
      481 gcgaatggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
      541 tatggtgcac tctcagtaca atctgctctg atgccgcata aggtctcagc ttttacttgt
      601 cttctaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
      661 ctcccggcat ccgcttacag acaagctgtg acccacgaag tgagtggacg aaagggcctc
      721 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
      781 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
      841 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
      901 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
      961 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1021 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1081 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1141 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1201 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1261 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1321 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1381 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1441 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1501 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1561 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1621 ggttctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1681 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     1741 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     1801 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     1861 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1921 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1981 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2041 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     2101 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2161 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2221 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2281 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2341 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2401 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2461 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2521 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2581 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2641 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2701 gcggatgagc gcccaatacg caaac
//
