LOCUS       Exported                6983 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Bacterial expression of human 6His-tagged SUMO1 and SUMO-E1.
ACCESSION   .
VERSION     .
KEYWORDS    pSA1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6983)
  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 6983)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6983
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        549..567
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    568..592
                     /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)."
     primer_bind     complement(3667..3685)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3681..3728
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(3899..4690)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(4691..4782)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4830..5568)
                     /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."
     CDS             complement(5778..6860)
                     /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     6822..6841
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6861..6938)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt
       61 tgtttaactt taataaggag atataccatg ggcagcagcc atcaccatca tcaccacagc
      121 caggatctgg ttccgcgtat gtctgaccag gaggccaaac cttcaactga ggacttgggg
      181 gataagaagg aaggtgaata tattaaactc aaagtcattg gacaggatag cagtgagatt
      241 cacttcaaag tgaaaatgac aacacatctc aagaaactca aagaatcata ctgtcaaaga
      301 cagggtgttc caatgaattc actcaggttt ctctttgagg gtcagagaat tgctgataat
      361 catactccaa aagaactggg aatggaggaa gaagatgtga ttgaagttta tcaggaacaa
      421 acggggggtt agggatcccc gggaattcat cgtgactgac tgacgctgca ggtcgacaag
      481 cttgcggccg cataatgctt aagtcgaaca gaaagtaatc gtattgtaca cggccgcata
      541 atcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt ccccatctta
      601 gtatattagt taagtataag aaggagatat acatatggca gatctcaatt cccgggtcga
      661 catggcactg tcgcgggggc tgccccggga gctggctgag gcggtggccg ggggccgggt
      721 gctggtggtg ggggcgggcg gcatcggctg cgagctcctc aagaatctcg tgctcaccgg
      781 tttctcccac atcgacctga ttgatctgga tactattgat gtaagcaacc tcaacagaca
      841 gtttttgttt caaaagaaac atgttggaag atcaaaggca caggttgcca aggaaagtgt
      901 actgcagttt tacccgaaag ctaatatcgt tgcctaccat gacagcatca tgaaccctga
      961 ctataatgtg gaatttttcc gacagtttat actggttatg aatgctttag ataacagagc
     1021 tgcccgaaac catgttaata gaatgtgcct ggcagctgat gttcctctta ttgaaagtgg
     1081 aacagctggg tatcttggac aagtaactac tatcaaaaag ggtgtgaccg agtgttatga
     1141 gtgtcatcct aagccgaccc agagaacctt tcctggctgt acaattcgta acacaccttc
     1201 agaacctata cattgcatcg tttgggcaaa gtacttgttc aaccagttgt ttggggaaga
     1261 agatgctgat caagaagtat ctcctgacag agctgaccct gaagctgcct gggaaccaac
     1321 ggaagccgaa gccagagcta gagcatctaa tgaagatggt gacattaaac gtatttctac
     1381 taaggaatgg gctaaatcaa ctggatatga tccagttaaa ctttttacca agctttttaa
     1441 agatgacatc aggtatctgt tgacaatgga caaactatgg cggaaaagga aacctccagt
     1501 tccgttggac tgggctgaag tacaaagtca aggagaagaa acgaatgcat cagatcaaca
     1561 gaatgaaccc cagttaggcc tgaaagacca gcaggttcta gatgtaaaga gctatgcacg
     1621 tcttttttca aagagcatcg agactttgag agttcattta gcagaaaagg gggatggagc
     1681 tgagctcata tgggataagg atgacccatc tgcaatggat tttgtcacct ctgctgcaaa
     1741 cctcaggatg catattttca gtatgaatat gaagagtaga tttgatatca aatcaatggc
     1801 agggaacatt attcctgcta ttgctactac taatgcagta attgctgggt tgatagtatt
     1861 ggaaggattg aagattttat caggaaaaat agaccagtgc agaacaattt ttttgaataa
     1921 acaaccaaac ccaagaaaga agcttcttgt gccttgtgca ctggatcctc ccaaccccaa
     1981 ttgttatgta tgtgccagca agccagaggt gactgtgcgg ctgaatgtcc ataaagtgac
     2041 tgttctcacc ttacaagaca agatagtgaa agaaaaattt gctatggtag caccagatgt
     2101 ccaaattgaa gatgggaaag gaacaatcct aatatcttcc gaagagggag agacggaagc
     2161 taataatcac aagaagttgt cagaatttgg aattagaaat ggcagccggc ttcaagcaga
     2221 tgacttcctc caggactata ctttattgat caacatcctt catagtgaag acctaggaaa
     2281 ggacgttgaa tttgaagttg ttggtgatgc tccggaaaaa gtggggccca aacaagctga
     2341 agatgctgcc aaaagcataa ccaatggcag tgatgatgga gctcagccct ccacctccac
     2401 agctcaagag caagatgacg ttctcatagt tgattcggat gaagaagatt cttcaaataa
     2461 tgccgacgtc agtgaagaag agagaagccg caagaggaaa ttagatgaga aagagaatct
     2521 cagtgcaaag aggtcacgta tagaacagaa ggaagagctt gatgatgtca tagcattaga
     2581 ttgagcggcc gcattaaaag gaggaattaa ctatggtgga gaaggaggag gctggcggcg
     2641 gcattagcga ggaggaggcg gcacagtatg accggcagat ccgcctgtgg ggactggagg
     2701 cccagaaacg gctgcgggcc tctcgggtgc ttcttgtcgg cttgaaagga cttggggctg
     2761 aaattgccaa gaatctcatc ttggcaggag tgaaaggact gaccatgctg gatcacgaac
     2821 aggtaactcc agaagatccc ggagctcagt tcttgattcg tactgggtct gttggccgaa
     2881 atagggctga agcctctttg gagcgagctc agaatctcaa ccccatggtg gatgtgaagg
     2941 tggacactga ggatatagag aagaaaccag agtcattttt cactcaattc gatgctgtgt
     3001 gtctgacttg ctgctccagg gatgtcatag ttaaagttga ccagatctgt cacaaaaata
     3061 gcatcaagtt ctttacagga gatgtttttg gctaccatgg atacacattt gccaatctag
     3121 gagagcatga gtttgtagag gagaaaacta aagttgccaa agttagccaa ggagtagaag
     3181 atgggcccga caccaagaga gcaaaacttg attcttctga gacaacgatg gtcaaaaaga
     3241 aggtggtctt ctgccctgtt aaagaagccc tggaggtgga ctggagcagt gagaaagcaa
     3301 aggctgctct gaagcgcacg acctccgact actttctcct tcaagtgctc ttaaagttcc
     3361 gtacagataa aggaagagat cccagttctg atacatatga ggaagattct gagttgttgc
     3421 tccagatacg aaatgatgtg cttgactcac tgggtattag tcctgacctg cttcctgagg
     3481 actttgtcag gtactgcttc tccgagatgg ccccagtgtg tgcggtggtt ggagggattt
     3541 tggcacagga aattgtgaag gccctgtctc agcgggaccc tcctcacaac aacttcttct
     3601 tcttcgatgg catgaagggg aatgggattg tggagtgcct tggccccaag ggctaggctg
     3661 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     3721 gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca
     3781 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga
     3841 cgaccgggtc atcgtggccg gatcttgcgg cccctcggct tgaacgaatt gttagacatt
     3901 atttgccgac taccttggtg atctcgcctt tcacgtagtg gacaaattct tccaactgat
     3961 ctgcgcgcga ggccaagcga tcttcttctt gtccaagata agcctgtcta gcttcaagta
     4021 tgacgggctg atactgggcc ggcaggcgct ccattgccca gtcggcagcg acatccttcg
     4081 gcgcgatttt gccggttact gcgctgtacc aaatgcggga caacgtaagc actacatttc
     4141 gctcatcgcc agcccagtcg ggcggcgagt tccatagcgt taaggtttca tttagcgcct
     4201 caaatagatc ctgttcagga accggatcaa agagttcctc cgccgctgga cctaccaagg
     4261 caacgctatg ttctcttgct tttgtcagca agatagccag atcaatgtcg atcgtggctg
     4321 gctcgaagat acctgcaaga atgtcattgc gctgccattc tccaaattgc agttcgcgct
     4381 tagctggata acgccacgga atgatgtcgt cgtgcacaac aatggtgact tctacagcgc
     4441 ggagaatctc gctctctcca ggggaagccg aagtttccaa aaggtcgttg atcaaagctc
     4501 gccgcgttgt ttcatcaagc cttacggtca ccgtaaccag caaatcaata tcactgtgtg
     4561 gcttcaggcc gccatccact gcggagccgt acaaatgtac ggccagcaac gtcggttcga
     4621 gatggcgctc gatgacgcca actacctctg atagttgagt cgatacttcg gcgatcaccg
     4681 cttccctcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4741 tgagcggata catatttgaa tgtatttaga aaaataaaca aatagctagc tcactcggtc
     4801 gctacgctcc gggcgtgaga ctgcggcggg cgctgcggac acatacaaag ttacccacag
     4861 attccgtgga taagcagggg actaacatgt gaggcaaaac agcagggccg cgccggtggc
     4921 gtttttccat aggctccgcc ctcctgccag agttcacata aacagacgct tttccggtgc
     4981 atctgtggga gccgtgaggc tcaaccatga atctgacagt acgggcgaaa cccgacagga
     5041 cttaaagatc cccaccgttt ccggcgggtc gctccctctt gcgctctcct gttccgaccc
     5101 tgccgtttac cggatacctg ttccgccttt ctcccttacg ggaagtgtgg cgctttctca
     5161 tagctcacac actggtatct cggctcggtg taggtcgttc gctccaagct gggctgtaag
     5221 caagaactcc ccgttcagcc cgactgctgc gccttatccg gtaactgttc acttgagtcc
     5281 aacccggaaa agcacggtaa aacgccactg gcagcagcca ttggtaactg ggagttcgca
     5341 gaggatttgt ttagctaaac acgcggttgc tcttgaagtg tgcgccaaag tccggctaca
     5401 ctggaaggac agatttggtt gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt
     5461 ccccaactga cttaaccttc gatcaaacca cctccccagg tggttttttc gtttacaggg
     5521 caaaagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctactgaa
     5581 ccgctctaga tttcagtgca atttatctct tcaaatgtag cacctgaagt cagccccata
     5641 cgatataagt tgtaattctc atgttagtca tgccccgcgc ccaccggaag gagctgactg
     5701 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
     5761 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     5821 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
     5881 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
     5941 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
     6001 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagatg
     6061 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
     6121 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
     6181 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
     6241 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
     6301 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
     6361 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
     6421 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
     6481 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
     6541 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
     6601 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
     6661 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
     6721 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
     6781 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
     6841 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
     6901 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
     6961 atgcgactcc tgcattagga aat
//
