LOCUS       sequence-382956-        5965 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 5965)
  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 5965)
  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-9 GFP(+6)-tag3
FEATURES             Location/Qualifiers
     source          1..5965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     58..75
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    90..229
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(155..174)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     315..337
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(331..522)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1094..2176)
                     /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     2138..2157
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2177..2254)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2460..2479
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2567..2586
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2567..2585
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2586..2610
                     /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             2625..2647
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2660..2677
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3497..3541
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3579..3597)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3593..3640
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3677..4132
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3764..3783)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3974..3995
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(4247..5038,5039..5107))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4870..4889
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5108..5200)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5281..5869
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5770..5789
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
       61 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
      121 tgcggtattt cacaccgcaa tggtgcactc tcagtacaat ctgctctgat gccgcatagt
      181 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
      241 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
      301 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
      361 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
      421 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
      481 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
      541 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
      601 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
      661 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
      721 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
      781 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
      841 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
      901 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
      961 cgacaggagc acgatcatgc tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     1021 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     1081 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1141 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     1201 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     1261 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     1321 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg
     1381 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     1441 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     1501 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     1561 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     1621 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     1681 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     1741 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     1801 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     1861 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     1921 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     1981 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     2041 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     2101 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     2161 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     2221 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     2281 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     2341 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     2401 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     2461 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     2521 atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg cgaaattaat acgactcact
     2581 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag
     2641 aaggagatat accatgggtc atcaccacca ccatcacggt ggcgctagca aaggtgaacg
     2701 tctgtttact ggtgtagtac cgatcttagt ggaattagac ggcgacgtga acggtcataa
     2761 atttagcgtg cgcggcgagg gcgaaggtga cgctaccaat ggtaaattga ccctgaagtt
     2821 tatttgcaca acaggcaaat tacccgttcc gtggcccacc ttagtgacca ccctgaccta
     2881 tggcgttcag tgcttcagtc gttaccctaa acatatgaaa caacacgatt ttttcaaatc
     2941 agccatgcct gaaggatatg ttcaagagcg tacaatcagc ttcaagaagg atggcaccta
     3001 taaaacgcgt gcggaagtga aatttgaagg ccgcacatta gtaaatcgta tcgaactgaa
     3061 aggtcgtgac ttcaaagaag acggcaacat tttaggccat aaactggaat ataactttaa
     3121 ttctcataac gtgtatatta cggccgataa acagaagaat ggtatcaagg caaatttcaa
     3181 aattcgccat aacgtggagg acggcagcgt tcaattagcg gatcattatc aacaaaacac
     3241 gccgattggt gatgggcctg tactgttacc tcgcaaccac tacctgagca cccaatctgc
     3301 cctgagcaaa gatccgaaag aaaaacgcga tcacatggtt ctgttagaat tcgtgaccgc
     3361 tgcaggcatt acgcacggaa tggacgaacg ctacaaggga ggaagcaaga aacgcaagaa
     3421 gcgtggcggt agcaaaaagc gtaagaagcg tggaggttca aagaaacgta aaaagcgata
     3481 ataactcgag tctggtaaag aaaccgctgc tgcgaaattt gaacgccagc acatggactc
     3541 gtctactagc gcagcttaat taacctaggc tgctgccacc gctgagcaat aactagcata
     3601 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     3661 cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     3721 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     3781 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     3841 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     3901 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     3961 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     4021 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     4081 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat ttctggcggc
     4141 acgatggcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4201 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4261 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4321 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4381 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4441 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4501 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4561 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4621 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4681 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4741 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4801 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4861 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4921 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4981 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5041 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5101 tactcatact cttccttttt caatcatgat tgaagcattt atcagggtta ttgtctcatg
     5161 agcggataca tatttgaatg tatttagaaa aataaacaaa taggtcatga ccaaaatccc
     5221 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5281 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5341 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5401 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     5461 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5521 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5581 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5641 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5701 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5761 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5821 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5881 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5941 gttatcccct gattctgtgg ataac
//
