LOCUS       sequence-205578-        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-TP1
FEATURES             Location/Qualifiers
     source          1..2725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    141..162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(177..182,183..200,201..207)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    215..231
                     /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     220..242
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     239..255
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     239..255
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             251..574
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    268..324
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(325..342)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(325..341)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(334..356)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(550..569)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(709..727)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        795..899
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(900..968,969..1760)
                     /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(1118..1137)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1931..2519
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2420..2439
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2673..2690
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccccgcgcgt tggccgattc attaatcact ctgtggttca gaagacattg ccttaacttt
       61 aagaaggaga tataccatgg agaccgcagc tggcacgaca ggtttcccga ctggaaagcg
      121 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
      181 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
      241 ggaaacagct atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga
      301 tccccgggta ccgagctcga attcactggc cgtcgtttta caacgtcgtg actgggaaaa
      361 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
      421 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
      481 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg
      541 cactctcagt acaatctgct ctgatgccgc ataaggtctc agcttgcaat tgtcttctaa
      601 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
      661 catccgctta cagacaagct gtgacccacg aagtgagtgg acgaaagggc ctcgtgatac
      721 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
      781 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
      841 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      901 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
      961 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     1021 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     1081 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     1141 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     1201 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     1261 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     1321 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     1381 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     1441 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     1501 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     1561 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtggttctc
     1621 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     1681 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     1741 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     1801 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1861 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1921 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1981 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     2041 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     2101 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     2161 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     2221 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     2281 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     2341 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     2401 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2461 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     2521 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     2581 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     2641 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggatg
     2701 agcgcccaat acgcaaaccg cctct
//
