LOCUS       sequence-245941-        5114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of RFP-SZ2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5114)
  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 5114)
  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_RFP-SZ2
FEATURES             Location/Qualifiers
     source          1..5114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     106..125
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        106..124
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    125..149
                     /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             164..186
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             197..214
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             932..952
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1115..1132
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1185..1203)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1199..1246
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1283..1738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1370..1389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1580..1601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1853..2668)
                     /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(1853..2668)
                     /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     2576..2595
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2660..2679
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2821..3409
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3310..3329
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3563..3580
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(3751..4833)
                     /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     4795..4814
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4834..4911)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
       61 acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata cgactcacta
      121 taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt aactttaaga
      181 aggagatata catatgcatc accaccatca tcatgtcagc aagggcgagg aggacaacat
      241 ggcgatcatc aaggagttta tgcggttcaa ggtgcacatg gaaggttctg tcaatggaca
      301 cgaatttgaa atcgagggtg agggggaagg acgcccatac gaggggaccc aaactgcgaa
      361 gcttaaagtt acaaaggggg gtccgttgcc gtttgcttgg gatatcttat caccgcagtt
      421 catgtatggc agcaaagctt acgtcaaaca cccagcagat attcctgatt atcttaaatt
      481 gtcgttcccc gaaggcttca agtgggaacg ggtaatgaac ttcgaagatg gaggggtagt
      541 tacggtcacc caggattctt ctctgcaaga tggcgagttc atttacaaag tgaaacttag
      601 aggcacaaat ttcccctctg atggacccgt tatgcagaag aaaacgatgg ggtgggaggc
      661 atctagtgag cgtatgtatc cagaagatgg ggctttaaaa ggggagatca aacagcgtct
      721 gaagttgaaa gacggcggtc attatgatgc cgaggtcaaa actacgtaca aggcaaaaaa
      781 gccagttcag ttgcctggtg cttacaacgt gaatatcaaa cttgatatca caagccacaa
      841 tgaggattat accattgttg aacaatatga aagagcagag ggccggcaca gtacaggcgg
      901 tatggatgag ttgtataaag gcggcggatc tgagaattta tatttccagg gcgctcgtaa
      961 tgcatacctg cgtaaaaaaa tcgcgcgttt aaaaaaagac aacttgcaac ttgagcgcga
     1021 cgaacaaaat ttggagaaaa taatcgccaa ccttcgggac gagatcgcgc gtctggaaaa
     1081 cgaggtggct tcgcatgagc aataatgact cgagcaccac caccaccacc actgagatcc
     1141 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     1201 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     1261 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1321 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1381 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1441 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1501 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1561 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1621 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1681 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
     1741 gtggcacttt tcggggtacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1801 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa
     1861 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     1921 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     1981 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     2041 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     2101 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     2161 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     2221 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     2281 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     2341 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     2401 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     2461 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     2521 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     2581 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc
     2641 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     2701 acagttttat tgttcatgac caaaatccct taacgtgacc taggtcatga ccaaaatccc
     2761 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2821 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2881 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2941 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     3001 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     3061 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     3121 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     3181 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     3241 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     3301 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     3361 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     3421 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     3481 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     3541 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgccataa
     3601 tccatgataa agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg
     3661 aaggagctga ctgggttgaa ggctctcaag ggcatcggtc gagatcccgg tgcctaatga
     3721 gtgagctaac ttacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3781 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3841 cgccagggtg gtttttcttt tcaccagtga gacgggcaac agctgattgc ccttcaccgc
     3901 ctggccctga gagagttgca gcaagcggtc cacgctggtt tgccccagca ggcgaaaatc
     3961 ctgtttgatg gtggttaacg gcgggatata acatgagctg tcttcggtat cgtcgtatcc
     4021 cactaccgag atatccgcac caacgcgcag cccggactcg gtaatggcgc gcattgcgcc
     4081 cagcgccatc tgatcgttgg caaccagcat cgcagtggga acgatgccct cattcagcat
     4141 ttgcatggtt tgttgaaaac cggacatggc actccagtcg ccttcccgtt ccgctatcgg
     4201 ctgaatttga ttgcgagtga gatatttatg ccagccagcc agacgcagac gcgccgagac
     4261 agaacttaat gggcccgcta acagcgcgat ttgctggtga cccaatgcga ccagatgctc
     4321 cacgcccagt cgcgtaccgt cttcatggga gaaaataata ctgttgatgg gtgtctggtc
     4381 agagacatca agaaataacg ccggaacatt agtgcaggca gcttccacag caatggcatc
     4441 ctggtcatcc agcggatagt taatgatcag cccactgacg cgttgcgcga gaagattgtg
     4501 caccgccgct ttacaggctt cgacgccgct tcgttctacc atcgacacca ccacgctggc
     4561 acccagttga tcggcgcgag atttaatcgc cgcgacaatt tgcgacggcg cgtgcagggc
     4621 cagactggag gtggcaacgc caatcagcaa cgactgtttg cccgccagtt gttgtgccac
     4681 gcggttggga atgtaattca gctccgccat cgccgcttcc actttttccc gcgttttcgc
     4741 agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc tgataagaga caccggcata
     4801 ctctgcgaca tcgtataacg ttactggttt cacattcacc accctgaatt gactctcttc
     4861 cgggcgctat catgccatac cgcgaaaggt tttgcgccat tcgatggtgt ccgggatctc
     4921 gacgctctcc cttatgcgac tcctgcatta ggaagcagcc cagtagtagg ttgaggccgt
     4981 tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat ggcgcccaac agtcccccgg
     5041 ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg aagtggcgag
     5101 cccgatcttc ccca
//
