LOCUS       sequence-342236-        3658 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 3658)
  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 3658)
  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-UbE2S
FEATURES             Location/Qualifiers
     source          1..3658
                     /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(937..961)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1040..1058)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1054..1101
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1198..1653
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1285..1304)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1495..1516
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1679..1783
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1784..1852,1853..2644)
                     /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(2002..2021)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2815..3403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3304..3323
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3557..3574
                     /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 tcaccatgaa ctccaacgtg gagaacctac ccccgcacat catccgcctg gtgtacaagg
      301 aggtgacgac actgaccgca gacccacccg atggcatcaa ggtctttccc aacgaggagg
      361 acctcaccga cctccaggtc accatcgagg gccctgaggg gaccccatat gctggaggtc
      421 tgttccgcat gaaactcctg ctggggaagg acttccctgc ctccccaccc aagggctact
      481 tcctgaccaa gatcttccac ccgaacgtgg gcgccaatgg cgagatctgc gtcaacgtgc
      541 tcaagaggga ctggacggct gagctgggca tccgacacgt actgctgacc atcaagtgcc
      601 tgctgatcca ccctaacccc gagtctgcac tcaacgagga ggcgggccgc ctgctcttgg
      661 agaactacga ggagtatgcg gctcgggccc gtctgctcac agagatccac gggggcgccg
      721 gcgggcccag cggcagggcc gaagccggtc gggccctggc cagtggcact gaagcttcct
      781 ccaccgaccc tggggcccca gggggcccgg gaggggctga gggtcccatg gccaagaagc
      841 atgctggcga gcgcgataag aagctggcgg ccaagaaaaa gacggacaag aagcgggcgc
      901 tgcggcggct gtagtcgacc aagggtgggc gcgccgaccc agctttcttg tacaaagtgg
      961 ttcgatctgg taccatggaa ttcgaagctt gatccggctg ctaacaaagc ccgaaaggaa
     1021 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     1081 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggatctgg cgtaatagcg
     1141 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgggacg
     1201 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1261 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1321 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1381 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1441 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1501 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1561 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     1621 cgaattttaa caaaatatta acgcttacaa tttaggtggc acttttcggg gaaatgtgcg
     1681 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1741 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1801 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1861 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1921 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     1981 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2041 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2101 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2161 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2221 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2281 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2341 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2401 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2461 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2521 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2581 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2641 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2701 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2761 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2821 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2881 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2941 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     3001 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3061 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3121 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3181 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3241 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3301 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3361 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3421 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3481 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3541 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     3601 accgcctctc cccgcgcgtt ggccgattca ttaatgcagg atctcgatcc cgcgaaat
//
