LOCUS       sequence-198482-        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"
     primer_bind     complement(112..130)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        198..302
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(303..371,372..1163)
                     /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(521..540)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1334..1922
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1823..1842
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2076..2093
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2269..2290
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2305..2310,2311..2328,2329..2335)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2343..2359
                     /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     2348..2370
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2367..2383
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2367..2383
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2379..2702
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2396..2452
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2453..2470)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2453..2469)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2462..2484)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2678..2697)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
       61 cggcatccgc ttacagacaa gctgtgaccc acgaagtgag tggacgaaag ggcctcgtga
      121 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
      181 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
      241 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
      301 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
      361 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
      421 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
      481 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
      541 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
      601 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
      661 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
      721 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
      781 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
      841 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
      901 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
      961 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtggtt
     1021 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     1081 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     1141 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     1201 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     1261 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     1321 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     1381 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     1441 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     1501 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     1561 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     1621 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     1681 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     1741 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     1801 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     1861 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     1921 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     1981 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     2041 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     2101 atgagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatcactct
     2161 gtggttcaga agacatcaga ttaactttaa gaaggagata taccatggag accgcagctg
     2221 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     2281 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     2341 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     2401 tgcatgcctg caggtcgact ctagaggatc cccgggtacc gagctcgaat tcactggccg
     2461 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     2521 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     2581 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     2641 tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat
     2701 aaggtctcag ctttgtgttg tcttc
//
