LOCUS       sequence-205591-        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-TP5
FEATURES             Location/Qualifiers
     source          1..2725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    80..101
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(116..121,122..139,140..146)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    154..170
                     /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     159..181
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     178..194
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     178..194
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             190..513
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    207..263
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(264..281)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(264..280)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(273..295)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(489..508)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(648..666)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        734..838
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(839..907,908..1699)
                     /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(1057..1076)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1870..2458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2359..2378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2612..2629
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agaaggagat ataccatgga gaccgcagct ggcacgacag gtttcccgac tggaaagcgg
       61 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
      121 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
      181 gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac tctagaggat
      241 ccccgggtac cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
      301 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      361 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      421 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
      481 actctcagta caatctgctc tgatgccgca taaggtctca gctttgtgtt gtcttctaag
      541 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
      601 atccgcttac agacaagctg tgacccacga agtgagtgga cgaaagggcc tcgtgatacg
      661 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
      721 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      781 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      841 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      901 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      961 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     1021 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     1081 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     1141 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     1201 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1261 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1321 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1381 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1441 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1501 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtggttctcg
     1561 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1621 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1681 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1741 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1801 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1861 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1921 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1981 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2041 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2101 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2161 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2221 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2281 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2341 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2401 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2461 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2521 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2581 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggatga
     2641 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat cactctgtgg
     2701 ttcagaagac atcagattaa cttta
//
