LOCUS       sequence-382958-        6071 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  fluorescent proteins with cationic peptide tags to promote 
            biomolecular condensate formation in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6071)
  AUTHORS   Yeong V, Wang JW, Horn JM, Obermeyer AC
  TITLE     Intracellular phase separation of globular proteins facilitated by 
            short cationic peptides.
  JOURNAL   Nat Commun. 2022 Dec 22;13(1):7882. doi: 10.1038/s41467-022-35529-2.
  PUBMED    36550144
REFERENCE   2  (bases 1 to 6071)
  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: "2022-12-22"; volume: "13(1):7882. doi"; pages: " 
            10.1038/s41467-022-35529-2"
COMMENT     Sequence Label: pVY-11 mScarletI-tag3
FEATURES             Location/Qualifiers
     source          1..6071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(937..2019)
                     /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     1981..2000
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2020..2097)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2303..2322
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2406..2425
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2406..2424
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2425..2449
                     /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             2465..2485
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2500..2517
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3411..3429)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3425..3472
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3509..3964
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3596..3615)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3806..3827
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4056..4871)
                     /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(4056..4871)
                     /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     4779..4798
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4863..4882
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4993..5581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5482..5501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5735..5752
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5767..5906
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5832..5851)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5992..6014
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(6008..6071,1..128))
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
       61 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
      121 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
      181 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
      241 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
      301 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
      361 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
      421 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
      481 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
      541 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
      601 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
      661 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
      721 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
      781 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
      841 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
      901 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
      961 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1021 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     1081 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     1141 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     1201 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     1261 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     1321 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     1381 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     1441 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     1501 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     1561 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     1621 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     1681 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     1741 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     1801 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     1861 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     1921 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     1981 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     2041 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     2101 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     2161 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     2221 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     2281 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     2341 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     2401 gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat
     2461 aattttgttt aactttaaga aggaggagta catatgggtc atcaccacca ccatcacggt
     2521 ggcgctagta aaggagaagc tgtgattaaa gagttcatgc gcttcaaagt tcacatggag
     2581 ggttctatga acggtcacga gttcgagatc gaaggcgaag gcgagggccg tccgtatgaa
     2641 ggcacccaga ccgccaaact gaaagtgact aaaggcggcc cgctgccttt ttcctgggac
     2701 atcctgagcc cgcaatttat gtacggttct agggcgttca tcaaacaccc agcggatatc
     2761 ccggactatt ataagcagtc ttttccggaa ggtttcaagt gggaacgcgt aatgaatttt
     2821 gaagatggtg gtgccgtgac cgtcactcag gacacctccc tggaggatgg caccctgatc
     2881 tataaagtta aactgcgtgg tactaatttt ccacctgatg gcccggtgat gcagaaaaag
     2941 acgatgggtt gggaggcgtc taccgaacgc ttgtatccgg aagatggtgt gctgaaaggc
     3001 gacattaaaa tggccctgcg cctgaaagat ggcggccgct atctggctga cttcaaaacc
     3061 acgtacaaag ccaagaaacc tgtgcagatg cctggcgcgt acaatgtgga ccgcaaactg
     3121 gacatcacct ctcataatga agattatacg gtggtagagc aatatgagcg ctccgagggt
     3181 cgtcattcta ccggtggcat ggatgaacta tacaaaggag gaagcaagaa acgcaagaag
     3241 cgtggcggta gcaaaaagcg taagaagcgt ggaggttcaa agaaacgtaa aaagcgataa
     3301 taaacttaat taaggatccg aattcgagct ccgtcgacaa gcttgcggcc gcactcgagc
     3361 agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca ccgctgagca
     3421 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     3481 aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg
     3541 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     3601 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3661 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     3721 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     3781 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     3841 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     3901 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca
     3961 atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4021 tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc gagcatcaaa
     4081 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     4141 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     4201 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     4261 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagt
     4321 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     4381 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     4441 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     4501 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     4561 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     4621 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     4681 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     4741 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     4801 tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt ttcccgttga
     4861 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     4921 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4981 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5041 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5101 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     5161 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5221 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5281 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5341 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5401 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5461 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5521 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5581 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5641 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     5701 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     5761 agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcaatgg
     5821 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     5881 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     5941 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     6001 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     6061 catcagcgtg g
//
