LOCUS       sequence-169517-        5866 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of RanBP2 SUMO E3 ligase fragment (aa 
            2304-3062) with C-terminal His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5866)
  AUTHORS   Werner A, Flotho A, Melchior F
  TITLE     The RanBP2/RanGAP1*SUMO1/Ubc9 complex is a multisubunit SUMO E3 
            ligase.
  JOURNAL   Mol Cell. 2012 May 11;46(3):287-98. doi: 
            10.1016/j.molcel.2012.02.017. Epub 2012 Mar 29.
  PUBMED    22464730
REFERENCE   2  (bases 1 to 5866)
  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"; doi: 
            "10.1016/j.molcel.2012.02.017"; journalName: "Mol Cell"; date: 
            "2012-05-11- 11"; volume: "46"; issue: "3"; pages: "287-98"
COMMENT     Sequence Label: RanBP2-RB3-4-pET23a
FEATURES             Location/Qualifiers
     source          1..5866
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        493..597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(598..666,667..1458)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(816..835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1629..2217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2118..2137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2371..2388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2403..2542
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2468..2487)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2628..2650
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2644..2835)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3344..3363
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3344..3362
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3394..3416
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5710..5727
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      5794..5841
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1141 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1741 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     1801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2341 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2401 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg caatggtgca
     2461 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     2521 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     2581 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     2641 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc
     2701 agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag
     2761 tttctccaga agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt
     2821 ttcctgtttg gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat
     2881 accgatgaaa cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt
     2941 actggaacgt tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat
     3001 cactcagggt caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca
     3061 gcagcatcct gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc
     3121 cagactttac gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt
     3181 tttgcagcag cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt
     3241 aaggcaaccc cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg
     3301 tggccaggac ccaacgctgc ccgagatctc gatcccgcga aattaatacg actcactata
     3361 gggagaccac aacggtttcc ctctagaaat aattttgttt aactttaaga aggagatata
     3421 catatggaga gagatggaca gtactttgaa cctgttgttc ctttacctga tctagttgaa
     3481 gtatccagtg gtgaggaaaa tgaacaagtt gtttttagtc acagggcaaa actctacaga
     3541 tatgataaag atgttggtca atggaaagaa aggggcattg gtgatataaa gattttacag
     3601 aattatgata ataagcaagt tcgtatagtg atgagaaggg accaagtatt aaaactttgt
     3661 gccaatcaca gaataactcc agacatgact ttgcaaaata tgaaagggac agaaagagta
     3721 tggttgtgga ctgcatgtga ttttgcagat ggagaaagaa aagtagagca tttagctgtt
     3781 cgttttaaac tacaggatgt tgcagactcg tttaagaaaa tttttgatga agcaaaaaca
     3841 gcccaggaaa aagattcttt gataacacct catgtttctc ggtcaagcac tcccagagag
     3901 tcaccatgtg gcaaaattgc tgtagctgta ttagaagaaa ccacaagaga gaggacagat
     3961 gttattcagg gtgatgatgt agcagatgca acttcagaag ttgaagtgtc tagcacatct
     4021 gaaacaacac caaaagcagt ggtttctcct ccaaagtttg tatttggttc agagtctgtt
     4081 aaaagcattt ttagtagtga aaaatcaaaa ccatttgcat tcggcaacag ttcagccact
     4141 gggtctttgt ttggatttag ttttaatgca cctttgaaaa gtaacaatag tgaaactagt
     4201 tcagtagccc agagtggatc tgaaagcaaa gtggaaccta aaaaatgtga actgtcaaag
     4261 aactctgata tcgaacagtc ttcagatagc aaagtcaaaa atctctttgc ttcctttcca
     4321 acggaagaat cttcaatcaa ctacacattt aaaacaccag aaaaggcaaa agagaagaaa
     4381 aaacctgaag attctccctc agatgatgat gttctcattg tatatgaact aactccaacc
     4441 gctgagcaga aagcccttgc aaccaaactt aaacttcctc caactttctt ctgctacaag
     4501 aatagaccag attatgttag tgaagaagag gaggatgatg aagatttcga aacagctgtc
     4561 aagaaactta atggaaaact atatttggat ggctcagaaa aatgtagacc cttggaagaa
     4621 aatacagcag ataatgagaa agaatgtatt attgtttggg aaaagaaacc aacagttgaa
     4681 gagaaggcaa aagcagatac gttaaaactt ccacctacat ttttttgtgg agtctgtagt
     4741 gatactgatg aagacaatgg aaatggggaa gactttcaat cagagcttca aaaagttcag
     4801 gaagctcaaa aatctcagac agaagaaata actagcacaa ctgacagtgt atatacaggt
     4861 gggactgaag tgatggtacc ttctttctgt aaatctgaag aacctgattc tattaccaaa
     4921 tccattagtt caccatctgt ttcctctgaa actatggaca aacctgtaga tttgtcaact
     4981 agaaaggaaa ttgatacaga ttctacaagc caaggggaaa gcaagatagt ttcatttgga
     5041 tttggaagta gcacagggct ctcatttgca gacttggctt ccagtaattc tggagatttt
     5101 gcttttggtt ctaaagataa aaatttccaa tgggcaaata ctggagcagc tgtgtttgga
     5161 acacagtcag tcggaaccca gtcagccggt aaagttggtg aagatgaaga tggtagtgat
     5221 gaagaagtag ttcataatga agatatccat tttgaaccaa tagtgtcact accagaggta
     5281 gaagtaaaat ctggagaaga agatgaagaa attttgttta aagagagagc caaactttat
     5341 agatgggatc gggatgtcag tcagtggaag gagcgcggtg ttggagatat aaagattctt
     5401 tggcatacaa tgaagaatta ttaccggatc ttaatgagaa gagaccaggt ttttaaagtg
     5461 tgtgcaaacc acgttattac taaaacaatg gaattaaagc ccttaaatgt ttcaaataat
     5521 gctttagttt ggactgcctc agattatgct gatggagaag caaaagtaga acagcttgca
     5581 gtgagattta aaactaaaga agtagctgat tgtttcaaga aaacatttga agaatgtcag
     5641 cagaatttaa tgaaactcca gaaaggacat gtatcactgg cagcagaatt atcaaaggag
     5701 accctcgagc accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa
     5761 gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa
     5821 cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggat
//
