LOCUS       V001155                 7815 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V001155
VERSION     V001155
KEYWORDS    pSUMO1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7815)
  AUTHORS   Weber AR, Schuermann D, Schar P
  TITLE     Versatile Recombinant SUMOylation System for the Production of
            SUMO-Modified Protein.
  JOURNAL   PLoS One. 2014 Jul 9;9(7):e102157. doi:
            10.1371/journal.pone.0102157. eCollection 2014.
   PUBMED   25007328
REFERENCE   2  (bases 1 to 7815)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7815)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2014-07-9"; pages: "
            10.1371/journal.pone.0102157. eCollection 2014"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..20
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    21..45
                     /label="lac operator"
                     /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             60..82
                     /label="RBS"
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             101..118
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             140..430
                     /gene="SUMO1"
                     /label="Small ubiquitin-related modifier 1"
                     /note="Small ubiquitin-related modifier 1 from Sus scrofa.
                     Accession#: A7WLH8"
     promoter        579..597
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    598..622
                     /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)."
     CDS             665..1138
                     /gene="ube2i"
                     /label="SUMO-conjugating enzyme UBC9"
                     /note="SUMO-conjugating enzyme UBC9 from Xenopus
                     tropicalis. Accession#: Q28CQ4"
     CDS             1154..1198
                     /label="S-Tag"
                     /note="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
     terminator      1250..1297
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     promoter        1382..1400
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1401..1425
                     /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)."
     CDS             1495..3414
                     /gene="UBA2"
                     /label="SUMO-activating enzyme subunit 2"
                     /note="SUMO-activating enzyme subunit 2 from Homo sapiens.
                     Accession#: Q9UBT2"
     CDS             3446..4483
                     /gene="SAE1"
                     /label="SUMO-activating enzyme subunit 1"
                     /note="SUMO-activating enzyme subunit 1 from Homo sapiens.
                     Accession#: Q9UBE0"
     terminator      4514..4561
                     /label="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA
                     polymerase"
     CDS             complement(4735..5523)
                     /label="SmR"
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     promoter        complement(5524..5615)
                     /label="AmpR promoter"
     rep_origin      complement(5663..6401)
                     /direction=LEFT
                     /label="CloDF13 ori"
                     /note="Plasmids containing the CloDF13 (CDF) origin of
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     protein_bind    complement(6577..6598)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6614..7693)
                     /label="lacI"
                     /note="lac repressor"
     promoter        complement(7694..7771)
                     /label="lacI promoter"
ORIGIN
        1 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctgt agaaataatt
       61 ttgtttaact ttaataagga gatataccat gggcagcagc catcaccatc atcaccacag
      121 ccaggatctg gttccgcgta tgtctgacca ggaggccaaa ccttcaactg aggacttggg
      181 ggataagaag gaaggtgaat atattaaact caaagtcatt ggacaggata gcagtgagat
      241 tcacttcaaa gtgaaaatga caacacatct caagaaactc aaagaatcat actgtcaaag
      301 acagggtgtt ccaatgaatt cactcaggtt tctctttgag ggtcagagaa ttgctgataa
      361 tcatactcca aaagaactgg gaatggagga agaagatgtg attgaagttt atcaggaaca
      421 aacggggggt tagggatccc cgggaattca tcgtgactga ctgacgctgc aggtcgacaa
      481 gcttgcggcc gcataatgct taaccccttg gggcctctaa acgggtcttg aggggttttt
      541 tgcatgcatc gtattgtaca cggccgcata atcgaaatta atacgactca ctatagggga
      601 attgtgagcg gataacaatt ccccatctta gtatattagt taagtataag aaggagatat
      661 acatatgtcg gggatcgccc tcagcagact cgcccaggag aggaaagcat ggaggaaaga
      721 ccacccattt ggtttcgtgg ctgtcccaac aaaaaatccc gatggcacga tgaacctcat
      781 gaactgggag tgcgccattc cgggaaagaa agggactccg tgggaaggag gcttgtttaa
      841 actacggatg cttttcaaag atgattatcc atcttcgcca ccaaaatgta aattcgaacc
      901 accattattt cacccgaatg tgtacccttc ggggacagtg tgcctgtcca tcttagagga
      961 ggacaaggac tggaggccag ccatcacaat caaacagatc ctattaggaa tacaggaact
     1021 tctaaatgaa ccaaatatcc aagacccagc tcaagcagag gcctacacga tttactgcca
     1081 aaacagagtg gagtacgaga aaagggtccg agcacaagcc aagaagtttg cgccctcata
     1141 actcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca tggactcgtc
     1201 tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac tagcataacc
     1261 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg catttgagaa
     1321 gcacggtacc attaagtcga acagaaagta atcgtattgt acacggccgc ataatcgaaa
     1381 ttaatacgac tcactatagg ggaattgtga gcggataaca attccccatc ttagtatatt
     1441 agttaagtat aagaaggaga tatacatatg gcagatctca attcccgggt cgacatggca
     1501 ctgtcgcggg ggctgccccg ggagctggct gaggcggtgg ccgggggccg ggtgctggtg
     1561 gtgggggcgg gcggcatcgg ctgcgagctc ctcaagaatc tcgtgctcac cggtttctcc
     1621 cacatcgacc tgattgatct ggatactatt gatgtaagca acctcaacag acagtttttg
     1681 tttcaaaaga aacatgttgg aagatcaaag gcacaggttg ccaaggaaag tgtactgcag
     1741 ttttacccga aagctaatat cgttgcctac catgacagca tcatgaaccc tgactataat
     1801 gtggaatttt tccgacagtt tatactggtt atgaatgctt tagataacag agctgcccga
     1861 aaccatgtta atagaatgtg cctggcagct gatgttcctc ttattgaaag tggaacagct
     1921 gggtatcttg gacaagtaac tactatcaaa aagggtgtga ccgagtgtta tgagtgtcat
     1981 cctaagccga cccagagaac ctttcctggc tgtacaattc gtaacacacc ttcagaacct
     2041 atacattgca tcgtttgggc aaagtacttg ttcaaccagt tgtttgggga agaagatgct
     2101 gatcaagaag tatctcctga cagagctgac cctgaagctg cctgggaacc aacggaagcc
     2161 gaagccagag ctagagcatc taatgaagat ggtgacatta aacgtatttc tactaaggaa
     2221 tgggctaaat caactggata tgatccagtt aaacttttta ccaagctttt taaagatgac
     2281 atcaggtatc tgttgacaat ggacaaacta tggcggaaaa ggaaacctcc agttccgttg
     2341 gactgggctg aagtacaaag tcaaggagaa gaaacgaatg catcagatca acagaatgaa
     2401 ccccagttag gcctgaaaga ccagcaggtt ctagatgtaa agagctatgc acgtcttttt
     2461 tcaaagagca tcgagacttt gagagttcat ttagcagaaa agggggatgg agctgagctc
     2521 atatgggata aggatgaccc atctgcaatg gattttgtca cctctgctgc aaacctcagg
     2581 atgcatattt tcagtatgaa tatgaagagt agatttgata tcaaatcaat ggcagggaac
     2641 attattcctg ctattgctac tactaatgca gtaattgctg ggttgatagt attggaagga
     2701 ttgaagattt tatcaggaaa aatagaccag tgcagaacaa tttttttgaa taaacaacca
     2761 aacccaagaa agaagcttct tgtgccttgt gcactggatc ctcccaaccc caattgttat
     2821 gtatgtgcca gcaagccaga ggtgactgtg cggctgaatg tccataaagt gactgttctc
     2881 accttacaag acaagatagt gaaagaaaaa tttgctatgg tagcaccaga tgtccaaatt
     2941 gaagatggga aaggaacaat cctaatatct tccgaagagg gagagacgga agctaataat
     3001 cacaagaagt tgtcagaatt tggaattaga aatggcagcc ggcttcaagc agatgacttc
     3061 ctccaggact atactttatt gatcaacatc cttcatagtg aagacctagg aaaggacgtt
     3121 gaatttgaag ttgttggtga tgctccggaa aaagtggggc ccaaacaagc tgaagatgct
     3181 gccaaaagca taaccaatgg cagtgatgat ggagctcagc cctccacctc cacagctcaa
     3241 gagcaagatg acgttctcat agttgattcg gatgaagaag attcttcaaa taatgccgac
     3301 gtcagtgaag aagagagaag ccgcaagagg aaattagatg agaaagagaa tctcagtgca
     3361 aagaggtcac gtatagaaca gaaggaagag cttgatgatg tcatagcatt agattgagcg
     3421 gccgcattaa aaggaggaat taactatggt ggagaaggag gaggctggcg gcggcattag
     3481 cgaggaggag gcggcacagt atgaccggca gatccgcctg tggggactgg aggcccagaa
     3541 acggctgcgg gcctctcggg tgcttcttgt cggcttgaaa ggacttgggg ctgaaattgc
     3601 caagaatctc atcttggcag gagtgaaagg actgaccatg ctggatcacg aacaggtaac
     3661 tccagaagat cccggagctc agttcttgat tcgtactggg tctgttggcc gaaatagggc
     3721 tgaagcctct ttggagcgag ctcagaatct caaccccatg gtggatgtga aggtggacac
     3781 tgaggatata gagaagaaac cagagtcatt tttcactcaa ttcgatgctg tgtgtctgac
     3841 ttgctgctcc agggatgtca tagttaaagt tgaccagatc tgtcacaaaa atagcatcaa
     3901 gttctttaca ggagatgttt ttggctacca tggatacaca tttgccaatc taggagagca
     3961 tgagtttgta gaggagaaaa ctaaagttgc caaagttagc caaggagtag aagatgggcc
     4021 cgacaccaag agagcaaaac ttgattcttc tgagacaacg atggtcaaaa agaaggtggt
     4081 cttctgccct gttaaagaag ccctggaggt ggactggagc agtgagaaag caaaggctgc
     4141 tctgaagcgc acgacctccg actactttct ccttcaagtg ctcttaaagt tccgtacaga
     4201 taaaggaaga gatcccagtt ctgatacata tgaggaagat tctgagttgt tgctccagat
     4261 acgaaatgat gtgcttgact cactgggtat tagtcctgac ctgcttcctg aggactttgt
     4321 caggtactgc ttctccgaga tggccccagt gtgtgcggtg gttggaggga ttttggcaca
     4381 ggaaattgtg aaggccctgt ctcagcggga ccctcctcac aacaacttct tcttcttcga
     4441 tggcatgaag gggaatggga ttgtggagtg ccttggcccc aagggctagg ctgctgccac
     4501 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     4561 gctgaaacct caggcatttg agaagcacac ggtcacactg cttccggtag tcaataaacc
     4621 ggtaaaccag caatagacat aagcggctat ttaacgaccc tgccctgaac cgacgaccgg
     4681 gtcatcgtgg ccggatcttg cggcccctcg gcttgaacga attgttagac attatttgcc
     4741 gactaccttg gtgatctcgc ctttcacgta gtggacaaat tcttccaact gatctgcgcg
     4801 cgaggccaag cgatcttctt cttgtccaag ataagcctgt ctagcttcaa gtatgacggg
     4861 ctgatactgg gccggcaggc gctccattgc ccagtcggca gcgacatcct tcggcgcgat
     4921 tttgccggtt actgcgctgt accaaatgcg ggacaacgta agcactacat ttcgctcatc
     4981 gccagcccag tcgggcggcg agttccatag cgttaaggtt tcatttagcg cctcaaatag
     5041 atcctgttca ggaaccggat caaagagttc ctccgccgct ggacctacca aggcaacgct
     5101 atgttctctt gcttttgtca gcaagatagc cagatcaatg tcgatcgtgg ctggctcgaa
     5161 gatacctgca agaatgtcat tgcgctgcca ttctccaaat tgcagttcgc gcttagctgg
     5221 ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg acttctacag cgcggagaat
     5281 ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg ttgatcaaag ctcgccgcgt
     5341 tgtttcatca agccttacgg tcaccgtaac cagcaaatca atatcactgt gtggcttcag
     5401 gccgccatcc actgcggagc cgtacaaatg tacggccagc aacgtcggtt cgagatggcg
     5461 ctcgatgacg ccaactacct ctgatagttg agtcgatact tcggcgatca ccgcttccct
     5521 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5581 atacatattt gaatgtattt agaaaaataa acaaatagct agctcactcg gtcgctacgc
     5641 tccgggcgtg agactgcggc gggcgctgcg gacacataca aagttaccca cagattccgt
     5701 ggataagcag gggactaaca tgtgaggcaa aacagcaggg ccgcgccggt ggcgtttttc
     5761 cataggctcc gccctcctgc cagagttcac ataaacagac gcttttccgg tgcatctgtg
     5821 ggagccgtga ggctcaacca tgaatctgac agtacgggcg aaacccgaca ggacttaaag
     5881 atccccaccg tttccggcgg gtcgctccct cttgcgctct cctgttccga ccctgccgtt
     5941 taccggatac ctgttccgcc tttctccctt acgggaagtg tggcgctttc tcatagctca
     6001 cacactggta tctcggctcg gtgtaggtcg ttcgctccaa gctgggctgt aagcaagaac
     6061 tccccgttca gcccgactgc tgcgccttat ccggtaactg ttcacttgag tccaacccgg
     6121 aaaagcacgg taaaacgcca ctggcagcag ccattggtaa ctgggagttc gcagaggatt
     6181 tgtttagcta aacacgcggt tgctcttgaa gtgtgcgcca aagtccggct acactggaag
     6241 gacagatttg gttgctgtgc tctgcgaaag ccagttacca cggttaagca gttccccaac
     6301 tgacttaacc ttcgatcaaa ccacctcccc aggtggtttt ttcgtttaca gggcaaaaga
     6361 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctact gaaccgctct
     6421 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
     6481 agttgtaatt ctcatgttag tcatgccccg cgcccaccgg aaggagctga ctgggttgaa
     6541 ggctctcaag ggcatcggtc gagatcccgg tgcctaatga gtgagctaac ttacattaat
     6601 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6661 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgccagggtg gtttttcttt
     6721 tcaccagtga gacgggcaac agctgattgc ccttcaccgc ctggccctga gagagttgca
     6781 gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg
     6841 gcgggatata acatgagctg tcttcggtat cgtcgtatcc cactaccgag atgtccgcac
     6901 caacgcgcag cccggactcg gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg
     6961 caaccagcat cgcagtggga acgatgccct cattcagcat ttgcatggtt tgttgaaaac
     7021 cggacatggc actccagtcg ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga
     7081 gatatttatg ccagccagcc agacgcagac gcgccgagac agaacttaat gggcccgcta
     7141 acagcgcgat ttgctggtga cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt
     7201 cttcatggga gaaaataata ctgttgatgg gtgtctggtc agagacatca agaaataacg
     7261 ccggaacatt agtgcaggca gcttccacag caatggcatc ctggtcatcc agcggatagt
     7321 taatgatcag cccactgacg cgttgcgcga gaagattgtg caccgccgct ttacaggctt
     7381 cgacgccgct tcgttctacc atcgacacca ccacgctggc acccagttga tcggcgcgag
     7441 atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc cagactggag gtggcaacgc
     7501 caatcagcaa cgactgtttg cccgccagtt gttgtgccac gcggttggga atgtaattca
     7561 gctccgccat cgccgcttcc actttttccc gcgttttcgc agaaacgtgg ctggcctggt
     7621 tcaccacgcg ggaaacggtc tgataagaga caccggcata ctctgcgaca tcgtataacg
     7681 ttactggttt cacattcacc accctgaatt gactctcttc cgggcgctat catgccatac
     7741 cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc gacgctctcc cttatgcgac
     7801 tcctgcatta ggaaa
//
