LOCUS       sequence-166596-        5870 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 5870)
  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 5870)
  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..5870
                     /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        494..598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(599..667,668..1459)
                     /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(817..836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1630..2218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2119..2138
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2372..2389
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2404..2546
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2472..2491)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2632..2654
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2648..2839)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     3348..3367
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3348..3366
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             3398..3420
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5714..5731
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      5798..5845
                     /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 attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
      601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
      661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
      721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
      781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
      841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
      901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
      961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg
     2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     3061 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     3121 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     3181 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     3241 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     3301 cccgtggcca ggacccaacg ctgcccgaga tctcgatccc gcgaaattaa tacgactcac
     3361 tatagggaga ccacaacggt ttccctctag aaataatttt gtttaacttt aagaaggaga
     3421 tatacatatg gagagagatg gacagtactt tgaacctgtt gttcctttac ctgatctagt
     3481 tgaagtatcc agtggtgagg aaaatgaaca agttgttttt agtcacaggg caaaactcta
     3541 cagatatgat aaagatgttg gtcaatggaa agaaaggggc attggtgata taaagatttt
     3601 acagaattat gataataagc aagttcgtat agtgatgaga agggaccaag tattaaaact
     3661 ttgtgccaat cacagaataa ctccagacat gactttgcaa aatatgaaag ggacagaaag
     3721 agtatggttg tggactgcat gtgattttgc agatggagaa agaaaagtag agcatttagc
     3781 tgttcgtttt aaactacagg atgttgcaga ctcgtttaag aaaatttttg atgaagcaaa
     3841 aacagcccag gaaaaagatt ctttgataac acctcatgtt tctcggtcaa gcactcccag
     3901 agagtcacca tgtggcaaaa ttgctgtagc tgtattagaa gaaaccacaa gagagaggac
     3961 agatgttatt cagggtgatg atgtagcaga tgcaacttca gaagttgaag tgtctagcac
     4021 atctgaaaca acaccaaaag cagtggtttc tcctccaaag tttgtatttg gttcagagtc
     4081 tgttaaaagc atttttagta gtgaaaaatc aaaaccattt gcattcggca acagttcagc
     4141 cactgggtct ttgtttggat ttagttttaa tgcacctttg aaaagtaaca atagtgaaac
     4201 tagttcagta gcccagagtg gatctgaaag caaagtggaa cctaaaaaat gtgaactgtc
     4261 aaagaactct gatatcgaac agtcttcaga tagcaaagtc aaaaatctct ttgcttcctt
     4321 tccaacggaa gaatcttcaa tcaactacac atttaaaaca ccagaaaagg caaaagagaa
     4381 gaaaaaacct gaagattctc cctcagatga tgatgttctc attgtatatg aactaactcc
     4441 aaccgctgag cagaaagccc ttgcaaccaa acttaaactt cctccaactt tcttctgcta
     4501 caagaataga ccagattatg ttagtgaaga agaggaggat gatgaagatt tcgaaacagc
     4561 tgtcaagaaa cttaatggaa aactatattt ggatggctca gaaaaatgta gacccttgga
     4621 agaaaataca gcagataatg agaaagaatg tattattgtt tgggaaaaga aaccaacagt
     4681 tgaagagaag gcaaaagcag atacgttaaa acttccacct acattttttt gtggagtctg
     4741 tagtgatact gatgaagaca atggaaatgg ggaagacttt caatcagagc ttcaaaaagt
     4801 tcaggaagct caaaaatctc agacagaaga aataactagc acaactgaca gtgtatatac
     4861 aggtgggact gaagtgatgg taccttcttt ctgtaaatct gaagaacctg attctattac
     4921 caaatccatt agttcaccat ctgtttcctc tgaaactatg gacaaacctg tagatttgtc
     4981 aactagaaag gaaattgata cagattctac aagccaaggg gaaagcaaga tagtttcatt
     5041 tggatttgga agtagcacag ggctctcatt tgcagacttg gcttccagta attctggaga
     5101 ttttgctttt ggttctaaag ataaaaattt ccaatgggca aatactggag cagctgtgtt
     5161 tggaacacag tcagtcggaa cccagtcagc cggtaaagtt ggtgaagatg aagatggtag
     5221 tgatgaagaa gtagttcata atgaagatat ccattttgaa ccaatagtgt cactaccaga
     5281 ggtagaagta aaatctggag aagaagatga agaaattttg tttaaagaga gagccaaact
     5341 ttatagatgg gatcgggatg tcagtcagtg gaaggagcgc ggtgttggag atataaagat
     5401 tctttggcat acaatgaaga attattaccg gatcttaatg agaagagacc aggtttttaa
     5461 agtgtgtgca aaccacgtta ttactaaaac aatggaatta aagcccttaa atgtttcaaa
     5521 taatgcttta gtttggactg cctcagatta tgctgatgga gaagcaaaag tagaacagct
     5581 tgcagtgaga tttaaaacta aagaagtagc tgattgtttc aagaaaacat ttgaagaatg
     5641 tcagcagaat ttaatgaaac tccagaaagg acatgtatca ctggcagcag aattatcaaa
     5701 ggagaccctc gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa
     5761 ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc
     5821 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggat
//
