LOCUS       sequence-205592-        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-TP6
FEATURES             Location/Qualifiers
     source          1..2725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    142..163
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(178..183,184..201,202..208)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    216..232
                     /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     221..243
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     240..256
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     240..256
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             252..575
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    269..325
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(326..343)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(326..342)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(335..357)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(551..570)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(710..728)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        796..900
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(901..969,970..1761)
                     /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(1119..1138)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1932..2520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2421..2440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2674..2691
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tccccgcgcg ttggccgatt cattaatcac tctgtggttc agaagacatt gtgttaactt
       61 taagaaggag atataccatg gagaccgcag ctggcacgac aggtttcccg actggaaagc
      121 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
      181 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
      241 aggaaacagc tatgaccatg attacgccaa gcttgcatgc ctgcaggtcg actctagagg
      301 atccccgggt accgagctcg aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa
      361 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
      421 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
      481 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
      541 gcactctcag tacaatctgc tctgatgccg cataaggtct cagcttgagc ttgtcttcta
      601 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
      661 gcatccgctt acagacaagc tgtgacccac gaagtgagtg gacgaaaggg cctcgtgata
      721 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
      781 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
      841 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
      901 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
      961 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     1021 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     1081 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     1141 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     1201 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     1261 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     1321 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     1381 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     1441 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     1501 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     1561 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtggttct
     1621 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     1681 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     1741 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     1801 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     1861 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1921 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1981 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     2041 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     2101 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     2161 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     2221 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     2281 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     2341 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     2401 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     2461 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     2521 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     2581 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     2641 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggat
     2701 gagcgcccaa tacgcaaacc gcctc
//
