LOCUS       sequence-205468-        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-TP4
FEATURES             Location/Qualifiers
     source          1..2725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    152..173
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(188..193,194..211,212..218)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    226..242
                     /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     231..253
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     250..266
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     250..266
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             262..585
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    279..335
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(336..353)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(336..352)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(345..367)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(561..580)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(720..738)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        806..910
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(911..979,980..1771)
                     /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(1129..1148)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1942..2530
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2431..2450
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2684..2701
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaccgcctc tccccgcgcg ttggccgatt cattaatcac tctgtggttc agaagacatt
       61 tacttaactt taagaaggag atataccatg gagaccgcag ctggcacgac aggtttcccg
      121 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
      181 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      241 aatttcacac aggaaacagc tatgaccatg attacgccaa gcttgcatgc ctgcaggtcg
      301 actctagagg atccccgggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt
      361 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
      421 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
      481 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
      541 cgcatatggt gcactctcag tacaatctgc tctgatgccg cataaggtct cagcttcaga
      601 ttgtcttcta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
      661 tctgctcccg gcatccgctt acagacaagc tgtgacccac gaagtgagtg gacgaaaggg
      721 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
      781 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      841 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      901 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      961 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     1021 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     1081 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     1141 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     1201 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     1261 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     1321 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     1381 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1441 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1501 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1561 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     1621 gcgtggttct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1681 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1741 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1801 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1861 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1921 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1981 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2041 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     2101 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2161 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2221 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2281 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     2341 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     2401 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2461 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2521 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2581 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2641 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2701 ggaagcggat gagcgcccaa tacgc
//
