LOCUS       sequence-422381-        3707 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 3707)
  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 3707)
  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-UbE2R2
FEATURES             Location/Qualifiers
     source          1..3707
                     /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(986..1010)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1089..1107)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1103..1150
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1247..1702
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1334..1353)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1544..1565
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1728..1832
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1833..1901,1902..2693)
                     /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(2051..2070)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2864..3452
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3353..3372
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3606..3623
                     /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 tcaccatggc ccagcagcag atgaccagct cgcagaaggc cctgatgctc gagctgaaat
      301 ccctgcagga ggaaccggtg gagggcttcc ggatcaccct ggtggacgag tccgacctct
      361 acaactggga ggtggccatc ttcggacccc ccaacaccct ctacgaaggc ggctacttca
      421 aggcgcatat taaatttcct attgactacc cctattcacc acctaccttc agattcttga
      481 ccaaaatgtg gcaccccaac atttatgaga atggagatgt atgcatttcg attcttcatc
      541 cgcctgtaga tgacccacag agtggagaac tgccttctga aaggtggaat cctactcaga
      601 atgtgaggac tatcctatta agtgtaatct cactgcttaa tgagcccaac accttctccc
      661 cagccaatgt cgatgcttca gttatgttca ggaaatggag agacagtaaa ggaaaagaca
      721 aagaatatgc tgaaattatt aggaaacaag tttcagccac taaggccgaa gcagaaaagg
      781 atggagtgaa ggtccccaca accctggcgg aatactgcat caaaactaaa gtgccttcca
      841 atgacaacag ctcagatttg ctttacgacg acttgtatga tgacgacatt gatgatgaag
      901 atgaggagga ggaagatgcc gactgttatg atgatgatga ttctgggaat gaggagtcgt
      961 gagtcgacga agggtgggcg cgccgaccca gctttcttgt acaaagtggt tcgatctggt
     1021 accatggaat tcgaagcttg atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     1081 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     1141 gggttttttg ctgaaaggag gaactatatc cggatctggc gtaatagcga agaggcccgc
     1201 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
     1261 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     1321 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     1381 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     1441 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     1501 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     1561 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     1621 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     1681 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1741 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1801 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1861 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1921 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1981 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2041 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2101 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2161 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2221 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2281 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2341 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     2401 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     2461 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     2521 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     2581 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2641 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2701 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2761 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2821 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2881 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2941 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3001 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3061 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3121 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3181 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3241 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3301 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3361 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3421 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3481 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     3541 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3601 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     3661 ccgcgcgttg gccgattcat taatgcagga tctcgatccc gcgaaat
//
