LOCUS       sequence-137834-        3448 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 3448)
  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 3448)
  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-UbE2A
FEATURES             Location/Qualifiers
     source          1..3448
                     /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(727..751)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(830..848)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      844..891
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      988..1443
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1075..1094)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1285..1306
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1469..1573
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1574..1642,1643..2434)
                     /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(1792..1811)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2605..3193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3094..3113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3347..3364
                     /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 tcaccatgtc caccccggct cggcggcgcc tcatgcggga cttcaagagg ttgcaggagg
      301 atcctccagc cggagtcagc ggggctccgt ccgagaacaa cataatggtg tggaacgcgg
      361 tcattttcgg gcctgaaggg accccgtttg aggatggaac atttaaactt acaatagaat
      421 tcactgaaga atatccaaat aaaccaccta cagttagatt tgtctctaag atgttccatc
      481 caaatgtcta tgcagatggt agtatatgtc tggacatact tcagaaccgt tggagtccaa
      541 cctatgatgt gtcttccatt ctaacatcca tacagtctct gttggatgaa cccaatccca
      601 atagtccagc aaacagccag gctgctcagc tgtaccagga gaacaaacgg gaatatgaaa
      661 agcgtgtttc tgcaatagta gaacaaagct ggcgtgattg ttgactcgag aagggtgggc
      721 gcgccgaccc agctttcttg tacaaagtgg ttcgatctgg taccatggaa ttcgaagctt
      781 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
      841 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
      901 ggaactatat ccggatctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      961 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     1021 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     1081 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1141 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1201 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1261 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1321 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1381 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     1441 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     1501 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1561 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1621 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1681 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1741 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1801 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     1861 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1921 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     1981 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2041 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2101 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2161 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2221 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2281 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2341 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2401 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2461 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2521 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2581 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2641 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2701 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2761 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2821 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2881 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     2941 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3001 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3061 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3121 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3181 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3241 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3301 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3361 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3421 ttaatgcagg atctcgatcc cgcgaaat
//
