LOCUS       sequence-256670-        8611 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Bacterial expression plasmid for (SUMO)10 client to assess SUMO-SIM 
            scaffolds..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8611)
  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 8611)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: (SUMO)10
FEATURES             Location/Qualifiers
     source          1..8611
                     /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             36..65
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             69..89
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3222..3242
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3285..3302
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3355..3373)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3369..3416
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3453..3908
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3540..3559)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3750..3771
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3934..4038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4039..4107,4108..4899)
                     /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(4257..4276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5070..5658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5559..5578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5812..5829
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5844..5983
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5909..5928)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6069..6091
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6085..6276)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(7085..8167)
                     /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     8129..8148
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(8168..8245)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     8451..8470
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     8554..8573
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8554..8572
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    8573..8597
                     /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 tgggccatca ccatcaccat caccatcacc
       61 atcacggcga aaacctatac ttccagggat ccggtgggag ttggggaggg tctatgtcag
      121 aagagaagcc taaggagggc gtgaaaaccg agaatgacca cattaacttg aaggttgctg
      181 gacaggatgg gtccgtggtt cagttcaaaa ttaaacgcca cacaccgttg tcgaaactga
      241 tgaaagcgta ctgcgaacgc cagggtcttt ctatgcgtca gattcgcttc cgctttgatg
      301 gtcaaccgat taatgaaacg gacacgccgg cacagctgga gatggaggat gaagacacta
      361 tcgacgtatt tcagcagcag accgtcgtag ggggctccgg cggaagcggg gggtcgggag
      421 gttccatgag cgaagagaaa ccaaaagaag gagttaaaac cgaaaacgac catatcaacc
      481 tgaaggtcgc cggtcaggac ggctcagtcg tgcagttcaa gattaaacgt cataccccat
      541 taagtaagct gatgaaggcg tattgtgagc gccaaggctt gtcgatgcgc cagattcgtt
      601 ttcgctttga tggtcagcca atcaacgaaa ccgacacccc ggcgcagctg gaaatggaag
      661 atgaagatac catcgatgtg ttccaacagc aaacggttgt tggcggaagc ggcgggagcg
      721 gtggtagcgg gggttctatg tccgaagaaa aaccgaaaga gggtgtgaag actgaaaacg
      781 accacattaa tctgaaggtg gcgggccagg acggttccgt tgttcagttt aaaatcaaac
      841 gtcacacccc gctgtcgaaa ctgatgaaag cctattgcga acgtcaaggg ctgagcatgc
      901 gtcaaattcg ttttcgcttt gatggtcagc caatcaacga aaccgataca ccggcgcagc
      961 tggagatgga agacgaggac accattgacg tcttccagca acaaactgta gtgggtggct
     1021 ccggtggctc tggtgggtct ggtggctcga tgagtgagga aaaacccaaa gaaggcgtca
     1081 agactgagaa cgatcatatt aacctgaaag tcgcgggtca agatggttca gtggtgcagt
     1141 tcaaaattaa acgccacacc ccgctgtcaa agctgatgaa agcctattgt gagcgtcaag
     1201 gtctgtcgat gcgccagatc cgctttcgct ttgatggtca gccaatcaac gagacagaca
     1261 ctccggcaca actggagatg gaagatgaag acaccatcga tgtttttcaa cagcagaccg
     1321 tggtgggagg gagcggcggt agcgggggat caggtggtag tatgtctgag gaaaagccga
     1381 aggaaggggt aaagaccgaa aatgatcaca ttaatctgaa agttgcgggc caagatggta
     1441 gtgtagtgca atttaaaatc aagcgtcaca ccccgttatc taaactgatg aaagcatatt
     1501 gcgaacgcca aggcctgtct atgcgccaga ttcgctttcg ctttgacggg caaccaatta
     1561 atgaaacgga caccccggca caactggaaa tggaagacga ggatactatt gacgtgtttc
     1621 agcaacagac agtggtcggc ggctccggtg gatcaggtgg gagtggaggg tctatgtcag
     1681 aagagaagcc taaggagggc gtgaaaaccg agaatgacca cattaacttg aaggttgctg
     1741 gacaggatgg gtccgtggtt cagttcaaaa ttaaacgcca cacaccgttg tcgaaactga
     1801 tgaaagcgta ctgcgaacgc cagggtcttt ctatgcgtca gattcgcttc cgctttgatg
     1861 gtcaaccgat taatgaaacg gacacgccgg cacagctgga gatggaggat gaagacacta
     1921 tcgacgtatt tcagcagcag accgtcgtag ggggctccgg cggaagcggg gggtcgggag
     1981 gttccatgag cgaagagaaa ccaaaagaag gagttaaaac cgaaaacgac catatcaacc
     2041 tgaaggtcgc cggtcaggac ggctcagtcg tgcagttcaa gattaaacgt cataccccat
     2101 taagtaagct gatgaaggcg tattgtgagc gccaaggctt gtcgatgcgc cagattcgtt
     2161 ttcgctttga tggtcagcca atcaacgaaa ccgacacccc ggcgcagctg gaaatggaag
     2221 atgaagatac catcgatgtg ttccaacagc aaacggttgt tggcggaagc ggcgggagcg
     2281 gtggtagcgg gggttctatg tccgaagaaa aaccgaaaga gggtgtgaag actgaaaacg
     2341 accacattaa tctgaaggtg gcgggccagg acggttccgt tgttcagttt aaaatcaaac
     2401 gtcacacccc gctgtcgaaa ctgatgaaag cctattgcga acgtcaaggg ctgagcatgc
     2461 gtcaaattcg ttttcgcttt gatggtcagc caatcaacga aaccgataca ccggcgcagc
     2521 tggagatgga agacgaggac accattgacg tcttccagca acaaactgta gtgggtggct
     2581 ccggtggctc tggtgggtct ggtggctcga tgagtgagga aaaacccaaa gaaggcgtca
     2641 agactgagaa cgatcatatt aacctgaaag tcgcgggtca agatggttca gtggtgcagt
     2701 tcaaaattaa acgccacacc ccgctgtcaa agctgatgaa agcctattgt gagcgtcaag
     2761 gtctgtcgat gcgccagatc cgctttcgct ttgatggtca gccaatcaac gagacagaca
     2821 ctccggcaca actggagatg gaagatgaag acaccatcga tgtttttcaa cagcagaccg
     2881 tggtgggagg gagcggcggt agcgggggat caggtggtag tatgtctgag gaaaagccga
     2941 aggaaggggt aaagaccgaa aatgatcaca ttaatctgaa agttgcgggc caagatggta
     3001 gtgtagtgca atttaaaatc aagcgtcaca ccccgttatc taaactgatg aaagcatatt
     3061 gcgaacgcca aggcctgtct atgcgccaga ttcgctttcg ctttgacggg caaccaatta
     3121 atgaaacgga caccccggca caactggaaa tggaagacga ggatactatt gacgtgtttc
     3181 agcaacagac agtggtcggc ggctcctgtg gtggcagtct cgagaacctg tattttcagg
     3241 gccgtaaacg taaacgtaaa cgtaaacgta aaggctgagt cgagcaccac caccaccacc
     3301 actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
     3361 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
     3421 aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc
     3481 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     3541 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3601 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3661 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3721 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3781 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg
     3841 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct
     3901 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     3961 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4021 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4081 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     4141 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     4201 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     4261 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     4321 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     4381 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     4441 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     4501 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     4561 gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc
     4621 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     4681 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4741 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4801 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4861 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4921 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4981 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5041 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5101 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5161 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     5221 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5281 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5341 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5401 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5461 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5521 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5581 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5641 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5701 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5761 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5821 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5881 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     5941 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     6001 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     6061 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     6121 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     6181 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     6241 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     6301 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     6361 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     6421 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     6481 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     6541 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     6601 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     6661 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     6721 gagcacgatc atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc
     6781 gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa
     6841 taccgcaagc gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat
     6901 gacccagagc gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag
     6961 tgcggcgacg atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct
     7021 caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt
     7081 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     7141 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca
     7201 gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc
     7261 ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga
     7321 tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc
     7381 gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca
     7441 gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca
     7501 tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt
     7561 tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg
     7621 cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat
     7681 gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa
     7741 cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga
     7801 tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc
     7861 cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa
     7921 tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca
     7981 gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg
     8041 ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca
     8101 cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg
     8161 gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa
     8221 aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc
     8281 attaggaagc agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt
     8341 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
     8401 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
     8461 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
     8521 gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt
     8581 gagcggataa caattcccct ctagaaataa t
//
