LOCUS       sequence-82635-e        7416 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expression of full length untagged human RanGAP1 in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7416)
  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 7416)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pET11a-hRanGAP1
FEATURES             Location/Qualifiers
     source          1..7416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..48)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        116..220
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(221..289,290..1081)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(439..458)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1252..1840
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1741..1760
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1994..2011
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2026..2168
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2094..2113)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2254..2276
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2270..2461)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3773..4855)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4817..4836
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4856..4933)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5139..5158
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5242..5261
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5242..5260
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5261..5285
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             5300..5322
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6253..6270
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     primer_bind     complement(7143..7161)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7157..7204
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(7379..7384,7385..7401,7402..7407))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 acttggtaca ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
       61 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
      121 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      181 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
      241 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
      301 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
      361 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
      421 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
      481 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
      541 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
      601 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
      661 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
      721 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
      781 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
      841 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
      901 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
      961 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     1021 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1081 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     1141 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     1201 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1261 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1321 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1381 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1441 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1501 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1561 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     1621 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     1681 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     1741 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     1801 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     1861 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     1921 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     1981 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     2041 tctccttacg catctgtgcg gtatttcaca ccgcatatat ggtgcactct cagtacaatc
     2101 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
     2161 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     2221 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     2281 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
     2341 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
     2401 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
     2461 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
     2521 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
     2581 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
     2641 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
     2701 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
     2761 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
     2821 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
     2881 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa
     2941 cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct
     3001 gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg
     3061 gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
     3121 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca
     3181 tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc
     3241 agtgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc
     3301 gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc
     3361 gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta
     3421 gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     3481 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     3541 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     3601 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     3661 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     3721 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     3781 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     3841 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     3901 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     3961 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     4021 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
     4081 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     4141 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     4201 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     4261 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     4321 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     4381 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     4441 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     4501 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     4561 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     4621 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     4681 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     4741 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     4801 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     4861 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     4921 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     4981 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     5041 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     5101 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     5161 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     5221 gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     5281 attcccctct agaaataatt ttgtttaact ttaagaagga gatatacata tggcctcgga
     5341 agacattgcc aagctggcag agacacttgc caagactcag gtggccgggg gacagctgag
     5401 tttcaaaggc aagagcctca aactcaacac tgcagaagat gctaaagatg tgattaaaga
     5461 gattgaagac tttgacagct tggaggctct gcgtctggaa ggcaacacag tgggcgtgga
     5521 agccgccagg gtcatcgcca aggccttaga gaagaagtcg gagttgaagc gctgccactg
     5581 gagtgacatg ttcacgggaa ggctgcggac cgagatccca ccagccctga tctcactagg
     5641 ggaaggactc atcacagctg gggctcagct ggtggagctg gacttaagcg acaacgcatt
     5701 cgggcccgac ggtgtgcaag gcttcgaggc cctgctcaag agctcagcct gcttcaccct
     5761 gcaggaactc aagctcaaca actgtggcat gggcattggc ggcggcaaga tcctggctgc
     5821 agctctgacc gaatgtcacc ggaaatccag tgcccaaggc aagcctctgg ccctgaaggt
     5881 ctttgtggct ggcagaaacc gtctggagaa tgatggcgcc actgccttgg cagaagcttt
     5941 tagggtcatc gggaccctgg aggaggtcca catgccacag aatgggatca accaccctgg
     6001 catcactgcc ctggcccagg ctttcgctgt caaccccctg ctgcgggtca tcaacctgaa
     6061 tgacaacacc ttcactgaga agggcgccgt ggccatggcc gagaccttga agaccttgcg
     6121 gcaggtggag gtgattaatt ttggggactg cctggtgcgc tccaagggtg cagttgccat
     6181 tgcagatgcc atccgcggcg gcctgcccaa gctaaaggag ctgaacttgt cattctgtga
     6241 gatcaagagg gatgctgccc tggctgttgc tgaggccatg gcagacaaag ctgagctgga
     6301 gaagctggac ctgaatggca acaccctggg agaagaaggc tgtgaacagc ttcaggaggt
     6361 gctggagggc ttcaacatgg ccaaggtgct ggcgtccctc agtgatgacg aggacgagga
     6421 ggaggaggag gaaggagaag aggaagaaga ggaagcagaa gaagaggagg aggaagatga
     6481 ggaagaggag gaagaagagg aggaggagga ggaagaagag cctcagcagc gagggcaggg
     6541 agagaagtca gccacgccct cacggaagat tctggaccct aacactgggg agccagctcc
     6601 cgtgctgtcc tccccacctc ctgcagacgt ctccaccttc ctggcttttc cctctccaga
     6661 gaagctgctg cgcctagggc ccaagagctc cgtgctgata gcccagcaga ctgacacgtc
     6721 tgaccccgag aaggtggtct ctgccttcct aaaggtgtca tctgtgttca aggacgaagc
     6781 tactgtgagg atggcagtgc aggatgcagt agatgccctg atgcagaagg ctttcaactc
     6841 ctcgtccttc aactccaaca ccttcctcac caggctcctc gtgcacatgg gtctgctcaa
     6901 gagtgaagac aaggtcaagg ccattgccaa cctgtacggc cccctgatgg cgctgaacca
     6961 catggtgcag caggactatt tccccaaggc ccttgcaccc ctgctgctgg cgttcgtgac
     7021 caagcccaac agcgccctgg aatcctgctc cttcgcccgc cacagtctgc tgcagacgct
     7081 gtacaaggtc taggatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
     7141 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     7201 tttgctgaaa ggaggaacta tatccggata tcccgcaaga ggcccggcag taccggcata
     7261 accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg
     7321 ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat
     7381 taaagcttat cgatgataag ctgtcaaaca tgagaa
//
