LOCUS       sequence-382827-        3568 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 3568)
  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 3568)
  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-UbE2E3
FEATURES             Location/Qualifiers
     source          1..3568
                     /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"
     protein_bind    complement(847..871)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(950..968)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      964..1011
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1108..1563
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1195..1214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1405..1426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1589..1693
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1694..1762,1763..2554)
                     /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(1912..1931)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2725..3313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3214..3233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3467..3484
                     /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 acaaaaaagt tggcatgtcc agtgataggc
      241 aaaggtccga tgatgagagc cccagcacca gcagtggcag ttcagatgcg gaccagcgag
      301 acccagccgc tccagagcct gaagaacaag aggaaagaaa accttctgcc acccagcaga
      361 agaaaaacac caaactctct agcaaaacca ctgctaagtt atccactagt gctaaaagaa
      421 ttcagaagga gctagctgaa ataacccttg atcctcctcc taattgcagt gctgggccta
      481 aaggagataa catttatgaa tggagatcaa ctatacttgg tccaccgggt tctgtatatg
      541 aaggtggtgt gttttttctg gatatcacat tttcatcaga ttatccattt aagccaccaa
      601 aggttacttt ccgcaccaga atctatcact gcaacatcaa cagtcaggga gtcatctgtc
      661 tggacatcct taaagacaac tggagtcccg ctttgactat ttcaaaggtt ttgctgtcta
      721 tttgttccct tttgacagac tgcaaccctg cggatcctct ggttggaagc atagccactc
      781 agtatttgac caacagagca gaacacgaca ggatagccag acagtggacc aagagatacg
      841 caacataccc aactttcttg tacaaagtgg ttcgatctgg taccatggaa ttcgaagctt
      901 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
      961 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
     1021 ggaactatat ccggatctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     1081 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     1141 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     1201 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1261 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1321 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1381 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1441 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1501 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     1561 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1621 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1681 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1741 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1801 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1861 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1921 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1981 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     2041 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2101 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2161 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2221 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2281 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2341 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2401 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2461 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2521 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2581 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2641 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2701 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2761 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2821 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2881 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2941 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3001 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3061 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3121 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3181 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3241 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3301 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3361 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3421 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3481 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3541 ttaatgcagg atctcgatcc cgcgaaat
//
