LOCUS       sequence-156320-        3434 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 3434)
  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 3434)
  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: pET15-UbE2D1
FEATURES             Location/Qualifiers
     source          1..3434
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     23..42
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        23..41
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             73..95
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             115..132
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             136..168
                     /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(172..180,181..195)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     protein_bind    223..247
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(735..759)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(838..856)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      852..899
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      996..1451
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1083..1102)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1293..1314
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1477..1581
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1582..1650,1651..2442)
                     /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(1800..1819)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2613..3201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3102..3121
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3355..3372
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caggatctcg atcccgcgaa attaatacga ctcactatag ggagaccaca acggtttccc
       61 tctagaaata attttgttta actttaagaa ggagatatac atatgcgggg ttctcatcat
      121 catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg ggatctgtac
      181 gacgatgacg ataaggatcc gagctcgaga tctgcagatc aaacaagttt gtacaaaaaa
      241 gcaggctccg cggccgcccc cttcaccatg gcgctgaaga ggattcagaa agaattgagt
      301 gatctacagc gcgatccacc tgctcactgt tcagctggac ctgtgggaga tgacttgttc
      361 cactggcaag ccactattat ggggcctcct gatagcgcat atcaaggtgg agtcttcttt
      421 ctcactgtac attttccgac agattatcct tttaaaccac caaagattgc tttcacaaca
      481 aaaatttacc atccaaacat aaacagtaat ggaagtattt gtctcgatat tctgaggtca
      541 caatggtcac cagctctgac tgtatcaaaa gttttattgt ccatatgttc tctactttgt
      601 gatcctaatc cagatgaccc cttagtacca gatatcgcac aaatctataa atcagacaaa
      661 gaaaaataca acagacatgc aagagaatgg actcagaaat atgcaatgta actcgaggaa
      721 gggtgggcgc gccgacccag ctttcttgta caaagtggtt cgatctggta ccatggaatt
      781 cgaagcttga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
      841 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
      901 tgaaaggagg aactatatcc ggatctggcg taatagcgaa gaggcccgca ccgatcgccc
      961 ttcccaacag ttgcgcagcc tgaatggcga atgggacgcg ccctgtagcg gcgcattaag
     1021 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     1081 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     1141 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     1201 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     1261 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     1321 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     1381 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     1441 gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1501 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1561 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1621 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1681 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1741 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1801 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     1861 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1921 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1981 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2041 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2101 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     2161 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2221 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2281 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2341 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2401 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2461 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     2521 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     2581 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     2641 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     2701 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     2761 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     2821 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     2881 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     2941 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     3001 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     3061 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     3121 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     3181 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     3241 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     3301 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     3361 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     3421 ccgattcatt aatg
//
