LOCUS       sequence-389426-        3496 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 3496)
  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 3496)
  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-UbE2H
FEATURES             Location/Qualifiers
     source          1..3496
                     /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(775..799)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(878..896)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      892..939
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1036..1491
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1123..1142)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1333..1354
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1517..1621
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1622..1690,1691..2482)
                     /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(1840..1859)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2653..3241
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3142..3161
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3395..3412
                     /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 tggcatgtca tctcccagtc
      241 cgggcaagag gcggatggac acggacgtgg tcaagctcat cgagagtaaa catgaggtta
      301 cgatcctggg aggacttaat gaatttgtag tgaagtttta tggaccacaa ggaacaccat
      361 atgaaggcgg agtatggaaa gttagagtgg acctacctga taaataccct ttcaaatctc
      421 catctatagg attcatgaat aaaattttcc atcccaacat tgatgaagcg tcaggaactg
      481 tgtgtctaga tgtaattaat caaacttgga cagctctcta tgatcttacc aatatatttg
      541 agtccttcct gcctcagtta ttggcctatc ctaaccccat agatcctctc aatggtgacg
      601 ctgcagccat gtacctccac cgaccagaag aatacaagca gaaaattaaa gagtacatcc
      661 agaaatacgc cacggaggag gcgctgaaag aacaggaaga gggtaccggg gacagctcat
      721 cggagagctc tatgtctgac ttttccgaag atgaggccca ggatatggag ttgtacccaa
      781 ctttcttgta caaagtggtt cgatctggta ccatggaatt cgaagcttga tccggctgct
      841 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
      901 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
      961 ggatctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1021 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1081 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1141 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1201 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1261 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1321 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     1381 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1441 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     1501 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1561 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1621 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1681 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1741 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1801 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1861 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1921 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1981 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2041 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2101 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2161 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2221 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2281 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2341 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2401 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2461 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2521 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2581 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2641 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2701 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2761 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     2821 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     2881 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     2941 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3001 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3061 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3121 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3181 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcaga gcctatggaa
     3241 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3301 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3361 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3421 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcaggat
     3481 ctcgatcccg cgaaat
//
