LOCUS       sequence-205566-        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-P2
FEATURES             Location/Qualifiers
     source          1..2726
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     213..230
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    407..428
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(443..448,449..466,467..473)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    481..497
                     /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     486..508
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     505..521
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     505..521
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             517..840
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    534..590
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(591..608)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(591..607)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(600..622)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(816..835)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(975..993)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1061..1165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1166..1234,1235..2026)
                     /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(1384..1403)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(2197..2726,1..59)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2686..2705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
       61 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      121 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
      181 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggatg
      241 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tcactctgtg
      301 gttcagaaga catgcaatta actttaagaa ggagatatac aatgggagac cgcagctggc
      361 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      421 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      481 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      541 catgcctgca ggtcgactct agaggatccc cgggtaccga gctcgaattc actggccgtc
      601 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
      661 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
      721 cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg
      781 tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcataa
      841 ggtctcagct tactattgtc ttctaagcca gccccgacac ccgccaacac ccgctgacgc
      901 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga cccacgaagt
      961 gagtggacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     1021 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     1081 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1141 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1201 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1261 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1321 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1381 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     1441 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1501 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1561 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1621 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1681 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     1741 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     1801 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     1861 atctggagcc ggtgagcgtg gttctcgcgg tatcattgca gcactggggc cagatggtaa
     1921 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     1981 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2041 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2101 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2161 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2221 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2281 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2341 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2401 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2461 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2521 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2581 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2641 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2701 tctttatagt cctgtcgggt ttcgcc
//
