LOCUS       sequence-245543-        5378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SZ1-RGG-RGG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5378)
  AUTHORS   Schuster BS, Reed EH, Parthasarathy R, Jahnke CN, Caldwell RM, 
            Bermudez JG, Ramage H, Good MC, Hammer DA
  TITLE     Controllable protein phase separation and modular recruitment to 
            form responsive membraneless organelles.
  JOURNAL   Nat Commun. 2018 Jul 30;9(1):2985. doi: 10.1038/s41467-018-05403-1.
  PUBMED    30061688
REFERENCE   2  (bases 1 to 5378)
  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"; journalName: "Nat 
            Commun."; date: "2018-07-30"; volume: "9(1):2985. doi"; pages: " 
            10.1038/s41467-018-05403-1"
COMMENT     Sequence Label: pET_SZ1-RGG-RGG
FEATURES             Location/Qualifiers
     source          1..5378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     99..118
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        99..117
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    118..142
                     /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             157..179
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             844..861
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1372..1389
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1442..1460)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1456..1503
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1540..1995
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1627..1646)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1837..1858
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2110..2925)
                     /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(2110..2925)
                     /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     2833..2852
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2917..2936
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3078..3666
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3567..3586
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3820..3837
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(4008..5090)
                     /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     5052..5071
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5091..5168)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     join(5374..5378,1..15)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
       61 gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga
      121 attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta agaaggagat
      181 atacatatga acttggtagc ccagcttgag aacgaagttg cgagtctgga aaacgaaaac
      241 gagacgttga agaaaaagaa cttgcacaag aaagatttga ttgcctactt ggaaaaggaa
      301 attgccaatc tgcgtaaaaa gattgaggaa ggtatggaat ccaatcaatc caataacggt
      361 ggctcgggta atgccgctct gaatcgtggt ggccgctacg tgccgccgca tctgcgtggt
      421 ggcgatggtg gtgcagcagc agctgcatct gctggcggtg atgaccgtcg cggcggtgcg
      481 ggcggtggcg gttatcgtcg cggcggtggc aatagtggtg gcggtggcgg tggcggttat
      541 gatcgtggtt acaatgacaa ccgtgatgac cgcgataacc gtggcggttc cggcggttac
      601 ggtcgtgatc gtaattatga agaccgtggt tacaacggcg gtggcggtgg cggtggcaat
      661 cgtggctata acaataaccg tggtggcggt ggcggtggct acaatcgcca ggatcgtggt
      721 gacggtggca gctctaactt ttcgcgcggt ggctataata accgtgatga aggcagcgac
      781 aaccgcggta gtggccgttc ctacaataac gatcgtcgcg acaatggtgg cgatggcgaa
      841 ttcttagtac ctcgcgggtc aatggaatcc aatcaatcca ataacggtgg ctcgggtaat
      901 gccgctctga atcgtggtgg ccgctacgtg ccgccgcatc tgcgtggtgg cgatggtggt
      961 gcagcagcag ctgcatctgc tggcggtgat gaccgtcgcg gcggtgcggg cggtggcggt
     1021 tatcgtcgcg gcggtggcaa tagtggtggc ggtggcggtg gcggttatga tcgtggttac
     1081 aatgacaacc gtgatgaccg cgataaccgt ggcggttccg gcggttacgg tcgtgatcgt
     1141 aattatgaag accgtggtta caacggcggt ggcggtggcg gtggcaatcg tggctataac
     1201 aataaccgtg gtggcggtgg cggtggctac aatcgccagg atcgtggtga cggtggcagc
     1261 tctaactttt cgcgcggtgg ctataataac cgtgatgaag gcagcgacaa ccgcggtagt
     1321 ggccgttcct acaataacga tcgtcgcgac aatggtggcg atggcctcga gcaccaccac
     1381 caccaccact gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc
     1441 accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt
     1501 ttgctgaaag gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat
     1561 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     1621 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     1681 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     1741 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     1801 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     1861 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     1921 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     1981 taacgcttac aatttaggtg gcacttttcg gggtacccag gtggcacttt tcggggaaat
     2041 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2101 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     2161 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     2221 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     2281 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     2341 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     2401 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     2461 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     2521 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     2581 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     2641 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     2701 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     2761 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     2821 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     2881 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     2941 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgacctag
     3001 gtcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     3061 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     3121 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     3181 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     3241 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3301 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3361 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3421 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3481 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3541 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3601 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3661 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3721 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3781 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3841 gcggaagagc gccataatcc atgataaaga agacagtcat aagtgcggcg acgatagtca
     3901 tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc atcggtcgag
     3961 atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca ctgcccgctt
     4021 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     4081 gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac gggcaacagc
     4141 tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac gctggtttgc
     4201 cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca tgagctgtct
     4261 tcggtatcgt cgtatcccac taccgagata tccgcaccaa cgcgcagccc ggactcggta
     4321 atggcgcgca ttgcgcccag cgccatctga tcgttggcaa ccagcatcgc agtgggaacg
     4381 atgccctcat tcagcatttg catggtttgt tgaaaaccgg acatggcact ccagtcgcct
     4441 tcccgttccg ctatcggctg aatttgattg cgagtgagat atttatgcca gccagccaga
     4501 cgcagacgcg ccgagacaga acttaatggg cccgctaaca gcgcgatttg ctggtgaccc
     4561 aatgcgacca gatgctccac gcccagtcgc gtaccgtctt catgggagaa aataatactg
     4621 ttgatgggtg tctggtcaga gacatcaaga aataacgccg gaacattagt gcaggcagct
     4681 tccacagcaa tggcatcctg gtcatccagc ggatagttaa tgatcagccc actgacgcgt
     4741 tgcgcgagaa gattgtgcac cgccgcttta caggcttcga cgccgcttcg ttctaccatc
     4801 gacaccacca cgctggcacc cagttgatcg gcgcgagatt taatcgccgc gacaatttgc
     4861 gacggcgcgt gcagggccag actggaggtg gcaacgccaa tcagcaacga ctgtttgccc
     4921 gccagttgtt gtgccacgcg gttgggaatg taattcagct ccgccatcgc cgcttccact
     4981 ttttcccgcg ttttcgcaga aacgtggctg gcctggttca ccacgcggga aacggtctga
     5041 taagagacac cggcatactc tgcgacatcg tataacgtta ctggtttcac attcaccacc
     5101 ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt gcgccattcg
     5161 atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga agcagcccag
     5221 tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc
     5281 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
     5341 gagcccgaag tggcgagccc gatcttcccc atcggtga
//
