LOCUS       sequence-255200-        6325 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression plasmid for mCherry-(SIM)1 client to assess 
            SUMO-SIM scaffolds..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6325)
  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 6325)
  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: RFP-(SIM)1
FEATURES             Location/Qualifiers
     source          1..6325
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             75..104
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             108..128
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             132..839
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(277..295)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     545..564
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     739..762
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             975..995
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1038..1055
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1108..1126)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1122..1169
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1206..1661
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1293..1312)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1503..1524
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1687..1791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1792..1860,1861..2652)
                     /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(2010..2029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2823..3411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3312..3331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3565..3582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3597..3736
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3662..3681)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3822..3844
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3838..4029)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4838..5920)
                     /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     5882..5901
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5921..5998)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6204..6223
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6307..6325,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6307..6325
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatataccat gggccatcac catcaccatc accatcacca tcacggcgaa aacctatact
      121 tccagggatc catggtgagc aagggcgagg aggacaacat ggccatcatc aaggagttca
      181 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
      241 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggcg
      301 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
      361 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
      421 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
      481 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
      541 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
      601 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
      661 actacgacgc cgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
      721 cctacaacgt caacatcaag ctggacatca cctcccacaa cgaggactac accatcgtgg
      781 aacagtacga gcgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
      841 ccggaggtag tggcggtagc ggtggctctt ggggtgggag caaagtagat gtgatcgact
      901 tgaccatcga aagcagcagc gacgaagaag aagatccgcc tgccaaacgc ggcggtagct
      961 gcggtggctc tctcgagaac ctgtattttc agggccgtaa acgtaaacgt aaacgtaaac
     1021 gtaaaggctg agtcgagcac caccaccacc accactgaga tccggctgct aacaaagccc
     1081 gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg
     1141 cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattggcga
     1201 atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     1261 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     1321 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     1381 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     1441 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     1501 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     1561 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     1621 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     1681 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1741 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1801 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1861 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1921 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1981 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2041 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2101 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2161 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2221 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2281 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     2341 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2401 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2461 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2521 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2581 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2641 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2701 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2761 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2821 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     2881 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     2941 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3001 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3061 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3121 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3181 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3241 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3301 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3361 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3421 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3481 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3541 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3601 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca
     3661 gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga
     3721 ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     3781 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     3841 gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg
     3901 gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc
     3961 cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg
     4021 tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat
     4081 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     4141 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     4201 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca
     4261 tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact
     4321 ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca
     4381 gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca
     4441 accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc
     4501 cgccatgccg gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac
     4561 gaaggcttga gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt
     4621 cgcgctccag cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc
     4681 tacgagttgc atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc
     4741 ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc
     4801 ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     4861 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4921 tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct
     4981 tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc
     5041 gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt
     5101 cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca
     5161 ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat
     5221 tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg
     5281 ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg
     5341 ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca
     5401 gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg
     5461 tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa
     5521 tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa
     5581 gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca
     5641 cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt
     5701 gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt
     5761 gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg
     5821 ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac
     5881 cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac
     5941 tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg
     6001 ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     6061 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     6121 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     6181 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     6241 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
     6301 cgaaattaat acgactcact atagg
//
