LOCUS       Exported                7083 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Bacterial expression of GST-tagged TARGET proteins fused to SUMO-E2 
            with PreScission protease cleavage site linker.
ACCESSION   .
VERSION     .
KEYWORDS    pSC-PreE2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7083)
  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 7083)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7083
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        67..171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             172..1032
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERSPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(390..409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(1074..1587)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1269..1290)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1481..1500
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      1698..2286
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2187..2206
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2440..2457
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(2479..2498)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2639..2661
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2655..2846)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3403..4485)
                     /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     4447..4466
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4486..4563)
                     /gene="lacI"
                     /label=lacI promoter
     terminator      4716..4802
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4899..4985
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5082..5168
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5265..5351
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5448..5534
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     5669..5688
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5669..5687
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5688..5712
                     /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             5727..5749
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             5793..5816
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6324..6980
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     6935..6957
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     primer_bind     complement(7029..7047)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      7043..7
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ttttttgctg aaaggaggaa ctatatccgg ataactacgt caggtggcac ttttcgggga
       61 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
      121 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
      181 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
      241 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
      301 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
      361 tctccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
      421 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
      481 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
      541 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
      601 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
      661 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
      721 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
      781 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
      841 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
      901 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
      961 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     1021 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaccccgg
     1081 ttgataatca gaaaagcccc aaaaacagga agattgtata agcaaatatt taaattgtaa
     1141 acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     1201 aataggccga aatcggcaaa atcccttata aatcaaaaga atagcccgag atagggttga
     1261 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     1321 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc aaatcaagtt
     1381 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     1441 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     1501 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     1561 cgcttaatgc gccgctacag ggcgcgtaaa aggatctagg tgaagatcct ttttgataat
     1621 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     1681 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     1741 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     1801 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     1861 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     1921 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     1981 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2041 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2101 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2161 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2221 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2281 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2341 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2401 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     2461 gctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     2521 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     2581 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     2641 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     2701 taaagctcat cagcgtggtc gtgcagcgat tcacagatgt ctgcctgttc atccgcgtcc
     2761 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     2821 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     2881 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     2941 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     3001 cagagaaaaa tcactcaggg tcaatgccag ccgaacgcca gcaagacgta gcccagcgcg
     3061 tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt ggcgggacca
     3121 gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga caggccgatc
     3181 atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc tgccggcacc
     3241 tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat agtcatgccc
     3301 cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg tcgagatccc
     3361 ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc cgctttccag
     3421 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     3481 ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca acagctgatt
     3541 gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg tttgccccag
     3601 caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc tgtcttcggt
     3661 atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact cggtaatggc
     3721 gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc
     3781 ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt cgccttcccg
     3841 ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag ccagacgcag
     3901 acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt gacccaatgc
     3961 gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat
     4021 gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg cagcttccac
     4081 agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga cgcgttgcgc
     4141 gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta ccatcgacac
     4201 caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg
     4261 cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt tgcccgccag
     4321 ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt ccactttttc
     4381 ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg tctgataaga
     4441 gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca ccaccctgaa
     4501 ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc attcgatggt
     4561 gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta
     4621 ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgccggcat gccgcccttt
     4681 cgtcttcaag aattaattcc caattcccca ggcatcaaat aaaacgaaag gctcagtcga
     4741 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     4801 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     4861 gcccgccata aactgccagg aattaattcc ccaggcatca aataaaacga aaggctcagt
     4921 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga
     4981 caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag
     5041 gacgcccgcc ataaactgcc aggaattaat tccccaggca tcaaataaaa cgaaaggctc
     5101 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     5161 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     5221 caggacgccc gccataaact gccaggaatt aattccccag gcatcaaata aaacgaaagg
     5281 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     5341 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     5401 gggcaggacg cccgccataa actgccagga attaattccc caggcatcaa ataaaacgaa
     5461 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     5521 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     5581 ggcgggcagg acgcccgcca taaactgcca ggaattgggg atcggaatta attcccggtt
     5641 taaaccgggg atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
     5701 gataacaatt cccctctaga aataattttg tttaacttta agaaggagat ataaccatgg
     5761 ctagctcgcg agtcgacggc ggccgcgaat tcctcgaggt tctgttccag ggtccaggtg
     5821 gcggaagctt agcatccatg tcggggatcg ccctcagcag actcgcccag gagaggaaag
     5881 catggaggaa agaccaccca tttggtttcg tggctgtccc aacaaaaaat cccgatggca
     5941 cgatgaacct catgaactgg gagtgcgcca ttccgggaaa gaaagggact ccgtgggaag
     6001 gaggcttgtt taaactacgg atgcttttca aagatgatta tccatcttcg ccaccaaaat
     6061 gtaaattcga accaccatta tttcacccga atgtgtaccc ttcggggaca gtgtgcctgt
     6121 ccatcttaga ggaggacaag gactggaggc cagccatcac aatcaaacag atcctattag
     6181 gaatacagga acttctaaat gaaccaaata tccaagaccc agctcaagca gaggcctaca
     6241 cgatttactg ccaaaacaga gtggagtacg agaaaagggt ccgagcacaa gccaagaagt
     6301 ttgcgccctc atcaggatcc ggaatgtccc ctatactagg ttattggaaa attaagggcc
     6361 ttgtgcaacc cactcgactt cttttggaat atcttgaaga aaaatatgaa gagcatttgt
     6421 atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt tgaattgggt ttggagtttc
     6481 ccaatcttcc ttattatatt gatggtgatg ttaaattaac acagtctatg gccatcatac
     6541 gttatatagc tgacaagcac aacatgttgg gtggttgtcc aaaagagcgt gcagagattt
     6601 caatgcttga aggagcggtt ttggatatta gatacggtgt ttcgagaatt gcatatagta
     6661 aagactttga aactctcaaa gttgattttc ttagcaagct acctgaaatg ctgaaaatgt
     6721 tcgaagatcg tttatgtcat aaaacatatt taaatggtga tcatgtaacc catcctgact
     6781 tcatgttgta tgacgctctt gatgttgttt tatacatgga cccaatgtgc ctggatgcgt
     6841 tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat cccacaaatt gataagtact
     6901 tgaaatccag caagtatata gcatggcctt tgcagggctg gcaagccacg tttggtggtg
     6961 gcgaccatcc tccaaaatag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
     7021 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     7081 ggg
//
