LOCUS       sequence-342235-        3455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3455)
  AUTHORS   Jin J, Li X, Gygi SP, Harper JW
  TITLE     Dual E1 activation systems for ubiquitin differentially regulate E2 
            enzyme charging.
  JOURNAL   Nature. 2007 Jun 28. 447(7148):1135-8.
  PUBMED    17597759
REFERENCE   2  (bases 1 to 3455)
  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"; journalName: "Nature. 
            2007 Jun 28. 447(7148):1135-8."
COMMENT     Sequence Label: pDEST17-UbE2L3
FEATURES             Location/Qualifiers
     source          1..3455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             93..110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             114..146
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(150..158,159..173)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     protein_bind    201..225
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      244..253
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(734..758)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(837..855)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      851..898
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      995..1450
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1082..1101)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1292..1313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1476..1580
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1581..1649,1650..2441)
                     /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(1799..1818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2612..3200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3101..3120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3354..3371
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat ggtatggcta
      121 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatccga
      181 gctcgagatc tgcagatcaa acaagtttgt acaaaaaagc aggctccgcg gccgccccct
      241 tcaccatggc ggccagcagg aggctgatga aggagcttga agaaatccgc aaatgtggga
      301 tgaaaaactt ccgtaacatc caggttgatg aagctaattt attgacttgg caagggctta
      361 ttgttcctga caaccctcca tatgataagg gagccttcag aatcgaaatc aactttccag
      421 cagagtaccc attcaaacca ccgaagatca catttaaaac aaagatctat cacccaaaca
      481 tcgacgaaaa ggggcaggtc tgtctgccag taattagtgc cgaaaactgg aagccagcaa
      541 ccaaaaccga ccaagtaatc cagtccctca tagcactggt gaatgacccc cagcctgagc
      601 acccgcttcg ggctgaccta gctgaagaat actctaagga ccgtaaaaaa ttctgtaaga
      661 atgctgaaga gtttacaaag aaatatgggg aaaagcgacc tgtggactaa ctcgaggaag
      721 ggtgggcgcg ccgacccagc tttcttgtac aaagtggttc gatctggtac catggaattc
      781 gaagcttgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
      841 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
      901 gaaaggagga actatatccg gatctggcgt aatagcgaag aggcccgcac cgatcgccct
      961 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     1021 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1081 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1141 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1201 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1261 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1321 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     1381 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     1441 cttacaattt aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     1501 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     1561 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     1621 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     1681 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     1741 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     1801 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     1861 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     1921 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     1981 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     2041 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     2101 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     2161 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     2221 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     2281 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     2341 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     2401 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     2461 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     2521 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     2581 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     2641 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     2701 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     2761 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     2821 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2881 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2941 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     3001 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     3061 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     3121 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     3181 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     3241 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     3301 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     3361 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     3421 cgattcatta atgcaggatc tcgatcccgc gaaat
//
