LOCUS       sequence-232141-        8422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression vector for 10 sumo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8422)
  AUTHORS   Delarue M, Brittingham GP, Pfeffer S, Surovtsev IV, Pinglay S, 
            Kennedy KJ, Schaffer M, Gutierrez JI, Sang D, Poterewicz G, Chung 
            JK, Plitzko JM, Groves JT, Jacobs-Wagner C, Engel BD, Holt LJ
  TITLE     mTORC1 Controls Phase Separation and the Biophysical Properties of 
            the Cytoplasm by Tuning Crowding.
  JOURNAL   Cell. 2018 Jul 12;174(2):338-349.e20. doi: 
            10.1016/j.cell.2018.05.042. Epub 2018 Jun 21.
  PUBMED    29937223
REFERENCE   2  (bases 1 to 8422)
  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.2018.05.042"; journalName: "Cell"; date: 
            "2018-07-12- 12"; volume: "174"; issue: "2"; pages: "338-349.e20"
COMMENT     Sequence Label: pT7-HISx6-SUMOx10
FEATURES             Location/Qualifiers
     source          1..8422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(216..1298)
                     /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     1260..1279
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1299..1376)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1582..1601
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1689..1708
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1689..1707
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1708..1732
                     /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             1749..1771
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1797..1814
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4971..4988
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5041..5059)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5055..5102
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5139..5594
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5226..5245)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5436..5457
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5686..6501)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(5686..6501)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6409..6428
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6493..6512
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6623..7211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7112..7131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7365..7382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    7397..7536
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(7462..7481)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7622..7644
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(7638..7829)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc acctgtccta
       61 cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg ccccgcgccc
      121 accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat cccggtgcct
      181 aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
      241 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
      301 ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg attgcccttc
      361 accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc cagcaggcga
      421 aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc ggtatcgtcg
      481 tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat ggcgcgcatt
      541 gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat gccctcattc
      601 agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc ccgttccgct
      661 atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg cagacgcgcc
      721 gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa tgcgaccaga
      781 tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt gatgggtgtc
      841 tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc cacagcaatg
      901 gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg cgcgagaaga
      961 ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga caccaccacg
     1021 ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga cggcgcgtgc
     1081 agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc cagttgttgt
     1141 gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt ttcccgcgtt
     1201 ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata agagacaccg
     1261 gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct gaattgactc
     1321 tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat ggtgtccggg
     1381 atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     1441 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     1501 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     1561 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     1621 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatcg atctcgatcc
     1681 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt ccccttaatt
     1741 aaaataattt tgtttaactt taagaaggag atatacataa gcttatgggc agcagccatc
     1801 accatcatca ccacactagt ggttcatccg gatcttcagg gagttctggt tcaagcatgt
     1861 cagaagagaa gcctaaggag ggcgtgaaaa ccgagaatga ccacattaac ttgaaggttg
     1921 ctggacagga tgggtccgtg gttcagttca aaattaaacg ccacacaccg ttgtcgaaac
     1981 tgatgaaagc gtacagcgaa cgccagggtc tttctatgcg tcagattcgc ttccgctttg
     2041 atggtcaacc gattaatgaa acggacacgc cggcacagct ggagatggag gatgaagaca
     2101 ctatcgacgt atttcagcag cagaccgtcg tagggggctc cggcggaagc ggggggtcgg
     2161 gaggttccat gagcgaagag aaaccaaaag aaggagttaa aaccgaaaac gaccatatca
     2221 acctgaaggt cgccggtcag gacggctcag tcgtgcagtt caagattaaa cgtcataccc
     2281 cattaagtaa gctgatgaag gcgtatagtg agcgccaagg cttgtcgatg cgccagattc
     2341 gttttcgctt tgatggtcag ccaatcaacg aaaccgacac cccggcgcag ctggaaatgg
     2401 aagatgaaga taccatcgat gtgttccaac agcaaacggt tgttggcgga agcggcggga
     2461 gcggtggtag cgggggttct atgtccgaag aaaaaccgaa agagggtgtg aagactgaaa
     2521 acgaccacat taatctgaag gtggcgggcc aggacggttc cgttgttcag tttaaaatca
     2581 aacgtcacac cccgctgtcg aaactgatga aagcctatag cgaacgtcaa gggctgagca
     2641 tgcgtcaaat tcgttttcgc tttgatggtc agccaatcaa cgaaaccgat acaccggcgc
     2701 agctggagat ggaagacgag gacaccattg acgtcttcca gcaacaaact gtagtgggtg
     2761 gctccggtgg ctctggtggg tctggtggct cgatgagtga ggaaaaaccc aaagaaggcg
     2821 tcaagactga gaacgatcat attaacctga aagtcgcggg tcaagatggt tcagtggtgc
     2881 agttcaaaat taaacgccac accccgctgt caaagctgat gaaagcctat agtgagcgtc
     2941 aaggtctgtc gatgcgccag atccgctttc gctttgatgg tcagccaatc aacgagacag
     3001 acactccggc acaactggag atggaagatg aagacaccat cgatgttttt caacagcaga
     3061 ccgtggtggg agggagcggc ggtagcgggg gatcaggtgg tagtatgtct gaggaaaagc
     3121 cgaaggaagg ggtaaagacc gaaaatgatc acattaatct gaaagttgcg ggccaagatg
     3181 gtagtgtagt gcaatttaaa atcaagcgtc acaccccgtt atctaaactg atgaaagcat
     3241 atagcgaacg ccaaggcctg tctatgcgcc agattcgctt tcgctttgac gggcaaccaa
     3301 ttaatgaaac ggacaccccg gcacaactgg aaatggaaga cgaggatact attgacgtgt
     3361 ttcagcaaca gacagtcgta ggttcatccg gatcttcagg gagttctggt tcaagcatga
     3421 gcgaagagaa acctaaagaa ggagttaaaa ccgaaaatga ccatatcaac ctgaaggtgg
     3481 ccggccagga cggctcagtc gtgcagttca agatcaaacg tcatacccca ttaagtaagc
     3541 tgatgaaggc gtatagtgag cgccaaggct tgtcgatgcg ccagattcgt ttccgctttg
     3601 acggtcagcc aatcaacgaa accgacaccc cggcgcagct ggaaatggaa gacgaagata
     3661 ctatcgatgt gttccaacag caaacggtgg tgggcggctc cgggggaagc ggggggtccg
     3721 gaggtagtag atccatgtca gaagagaagc ctaaggaagg cgtgaaaacc gagaatgacc
     3781 acattaactt gaaggttgct ggacaggatg ggtccgtggt tcagttcaag attaaacgcc
     3841 atacaccgtt gtcgaaactg atgaaggcgt acagcgaacg tcagggtctt tctatgcgtc
     3901 agattcgctt ccgctttgat ggtcaaccga ttaatgaaac ggacacgccg gcacagctgg
     3961 agatggagga tgaggacact atcgacgtat ttcagcaaca gaccgttgtt ggatcttcag
     4021 gatcaagcgg ttcatctggt agttccatgt ccgaagaaaa accgaaggag ggtgtgaaga
     4081 ctgaaaacga ccatattaat ctgaaggtgg cgggccagga cggttccgtt gttcagttta
     4141 agatcaaacg ccacaccccg ctgtcgaaac tgatgaaagc ctatagtgaa cgtcaagggc
     4201 tgagcatgcg tcaaattcgt tttcgctttg atggtcaacc aatcaacgaa accgatacac
     4261 cggcgcagct ggaaatggaa gacgaggaca ccattgacgt cttccagcaa caaactgttg
     4321 ttggttcctc tggaagctca ggatctagtg ggtcatcaat gagtgaagaa aaacccaagg
     4381 aaggcgtcaa gactgagaac gatcatatta acctgaaagt cgcgggtcaa gatggttcag
     4441 tggtgcagtt taaaattaaa cgccacaccc cgttgtcaaa gctgatgaaa gcctatagtg
     4501 aacgtcaagg tctgtcgatg cgccagatcc gcttccgctt tgatggtcag ccaatcaacg
     4561 agacagacac tccggcacaa ctggagatgg aagatgaaga caccatcgat gttttccaac
     4621 agcagaccgt tgttggttca tcaggttcat ccggatctag cgggtcaagt atgtctgaag
     4681 aaaagccgaa ggaaggggta aagaccgaaa atgatcacat taatctgaaa gttgcgggcc
     4741 aggatggtag tgtagtgcaa tttaaaatta aacgtcacac cccgttgtct aaactgatga
     4801 aagcatatag cgaacgccaa ggcctgtcta tgcgccagat tcgctttcgc tttgacgggc
     4861 aaccaatcaa tgaaacggac accccggcac aactggaaat ggaagacgag gacactattg
     4921 acgtgtttca gcaacagaca tctagataaa ccggtgcggc cgcactcgag caccaccacc
     4981 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     5041 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     5101 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
     5161 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5221 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5281 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5341 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     5401 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     5461 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     5521 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     5581 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5641 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
     5701 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     5761 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     5821 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     5881 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     5941 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     6001 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     6061 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     6121 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     6181 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     6241 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     6301 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     6361 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     6421 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
     6481 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     6541 tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6601 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6661 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6721 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     6781 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6841 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6901 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6961 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     7021 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     7081 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     7141 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     7201 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     7261 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     7321 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     7381 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     7441 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     7501 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     7561 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     7621 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     7681 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     7741 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     7801 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     7861 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     7921 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     7981 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     8041 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     8101 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     8161 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     8221 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     8281 atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     8341 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     8401 agcgacaggc cgatcatcgt cg
//
