LOCUS       sequence-256126-        7711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for (SIM)10 client to assess SUMO-SIM 
            scaffolds..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7711)
  AUTHORS   Banani SF, Rice AM, Peeples WB, Lin Y, Jain S, Parker R, Rosen MK
  TITLE     Compositional Control of Phase-Separated Cellular Bodies.
  JOURNAL   Cell. 2016 Jul 28;166(3):651-663. doi: 10.1016/j.cell.2016.06.010. 
            Epub 2016 Jun 30.
  PUBMED    27374333
REFERENCE   2  (bases 1 to 7711)
  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.cell.2016.06.010"; journalName: "Cell"; date: 
            "2016-07-28- 28"; volume: "166"; issue: "3"; pages: "651-663"
COMMENT     Sequence Label: (SIM)10
FEATURES             Location/Qualifiers
     source          1..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             39..68
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             72..1172
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1029..1038
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1146..1169
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1221..1232
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1239..1259
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2322..2342
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2385..2402
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2455..2473)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2469..2516
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2553..3008
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2640..2659)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2850..2871
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3034..3138
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3139..3207,3208..3999)
                     /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(3357..3376)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4170..4758
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4659..4678
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4912..4929
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4944..5083
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5009..5028)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5169..5191
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5185..5376)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6185..7267)
                     /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     7229..7248
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7268..7345)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7551..7570
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     7654..7673
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7654..7672
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    7673..7697
                     /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)."
ORIGIN
        1 tttgtttaac tttaagaagg agatatacca tgtctggcca tcaccatcac catcaccatc
       61 accatcacgg catgaaaatc gaagaaggta aactggtaat ctggattaac ggcgataaag
      121 gctataacgg tctcgctgaa gtcggtaaga aattcgagaa agataccgga attaaagtca
      181 ccgttgagca tccggataaa ctggaagaga aattcccaca ggttgcggca actggcgatg
      241 gccctgacat tatcttctgg gcacacgacc gctttggtgg ctacgctcaa tctggcctgt
      301 tggctgaaat caccccggac aaagcgttcc aggacaagct gtatccgttt acctgggatg
      361 ccgtacgtta caacggcaag ctgattgctt acccgatcgc tgttgaagcg ttatcgctga
      421 tttataacaa agatctgctg ccgaacccgc caaaaacctg ggaagagatc ccggcgctgg
      481 ataaagaact gaaagcgaaa ggtaagagcg cgctgatgtt caacctgcaa gaaccgtact
      541 tcacctggcc gctgattgct gctgacgggg gttatgcgtt caagtatgaa aacggcaagt
      601 acgacattaa agacgtgggc gtggataacg ctggcgcgaa agcgggtctg accttcctgg
      661 ttgacctgat taaaaacaaa cacatgaatg cagacaccga ttactccatc gcagaagctg
      721 cctttaataa aggcgaaaca gcgatgacca tcaacggccc gtgggcatgg tccaacatcg
      781 acaccagcaa agtgaattat ggtgtaacgg tactgccgac cttcaagggt caaccatcca
      841 aaccgttcgt tggcgtgctg agcgcaggta ttaacgccgc cagtccgaac aaagagctgg
      901 caaaagagtt cctcgaaaac tatctgctga ctgatgaagg tctggaagcg gttaataaag
      961 acaaaccgct gggtgccgta gcgctgaagt cttacgagga agagttggcg aaagatccac
     1021 gtattgccgc caccatggaa aacgcccaga aaggtgaaat catgccgaac atcccgcaga
     1081 tgtccgcttt ctggtatgcc gtgcgtactg cggtgatcaa cgccgccagc ggtcgtcaga
     1141 ctgtcgatga agccctgaaa gacgcgcaga ctaattcgag ctcgaacaac aacaacaata
     1201 acaataacaa caacctcggg atcgagggaa ggatttcaga aaacctgtac ttccagggat
     1261 ccggtggctc ttggggtggg agcaaagtag atgtgatcga cttgaccatc gaaagcagca
     1321 gcgacgaaga agaagatccg cctgccaaac gcggcggtag cggtggctct ggcgggagcg
     1381 gcggctcgaa agttgatgtc attgacctta caattgaatc ttcgagcgat gaagaggaag
     1441 atcctccagc gaaacgtggc ggctcgggag gttcgggcgg cagcggcgga agcaaagtcg
     1501 atgtaattga cttaacgatc gaatctagca gcgatgaaga agaagatcca ccggctaaac
     1561 gtgggggctc agggggttct ggcggttcag gcggcagtaa agttgacgtg attgacctca
     1621 ccatcgaaag cagtagcgac gaggaagaag atccaccagc aaaacgcgga gggagcgggg
     1681 gcagcggcgg cagcggcgga tctaaagtgg acgttatcga tctgacgatc gaaagcagta
     1741 gcgatgagga agaggacccg ccggcgaaac ggggcggctc ggggggctca ggtggctctg
     1801 gtgggagcaa agtagatgtg atcgacttga ccatcgaaag cagcagcgac gaagaagaag
     1861 atccgcctgc caaacgcggc ggtagcggtg gctctggcgg gagcggcggc tcgaaagttg
     1921 atgtcattga ccttacaatt gaatcttcga gcgatgaaga ggaagatcct ccagcgaaac
     1981 gtggcggctc gggaggttcg ggcggcagcg gcggaagcaa agtcgatgta attgacttaa
     2041 cgatcgaatc tagcagcgat gaagaagaag atccaccggc taaacgtggg ggctcagggg
     2101 gttctggcgg ttcaggcggc agtaaagttg acgtgattga cctcaccatc gaaagcagta
     2161 gcgacgagga agaagatcca ccagcaaaac gcggagggag cgggggcagc ggcggcagcg
     2221 gcggatctaa agtggacgtt atcgatctga cgatcgaaag cagtagcgat gaggaagagg
     2281 acccgccggc gaaacggggc ggctcgtgcg ggggctcact cgagaacctg tattttcagg
     2341 gccgtaaacg taaacgtaaa cgtaaacgta aaggctgagt cgagcaccac caccaccacc
     2401 actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     2461 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     2521 aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     2581 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2641 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2701 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2761 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2821 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2881 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     2941 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     3001 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3061 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3121 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3181 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3241 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3301 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3361 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3421 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3481 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3541 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3601 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3661 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     3721 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3781 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3841 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3901 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3961 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4021 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4081 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4141 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4201 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4261 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     4321 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4381 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4441 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4501 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4561 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4621 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4681 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4741 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4801 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4861 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     4921 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     4981 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5041 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     5101 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5161 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5221 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     5281 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     5341 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     5401 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     5461 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     5521 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     5581 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     5641 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     5701 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     5761 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     5821 gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc
     5881 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     5941 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     6001 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     6061 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     6121 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     6181 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     6241 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
     6301 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
     6361 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     6421 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
     6481 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
     6541 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     6601 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     6661 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     6721 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
     6781 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     6841 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     6901 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     6961 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     7021 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     7081 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     7141 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     7201 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     7261 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     7321 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
     7381 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     7441 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     7501 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     7561 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     7621 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
     7681 gagcggataa caattcccct ctagaaataa t
//
