LOCUS       sequence-382887-        5911 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 5911)
  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 5911)
  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-7 GFP(+6)-tag1
FEATURES             Location/Qualifiers
     source          1..5911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     73..90
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    105..244
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(170..189)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     330..352
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(346..537)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1109..2191)
                     /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     2153..2172
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2192..2269)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2475..2494
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2582..2601
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2582..2600
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2601..2625
                     /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             2640..2662
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2675..2692
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3458..3502
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3540..3558)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3554..3601
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3638..4093
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3725..3744)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3935..3956
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(4208..4999,5000..5068))
                     /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     4831..4850
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5069..5161)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5242..5830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5731..5750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
       61 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
      121 ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct
      181 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
      241 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
      301 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
      361 gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga
      421 ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa
      481 tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga
      541 tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag
      601 gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg
      661 taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca
      721 gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca
      781 gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg
      841 gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct
      901 tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc
      961 tagccgggtc ctcaacgaca ggagcacgat catgctagtc atgccccgcg cccaccggaa
     1021 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     1081 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1141 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1201 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     1261 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     1321 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     1381 ctaccgagat gtccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     1441 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     1501 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     1561 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     1621 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     1681 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     1741 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     1801 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     1861 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     1921 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     1981 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     2041 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     2101 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     2161 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     2221 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     2281 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     2341 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     2401 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     2461 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     2521 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tcgatctcga tcccgcgaaa
     2581 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt
     2641 ttgtttaact ttaagaagga gatataccat gggtcatcac caccaccatc acggtggcgc
     2701 tagcaaaggt gaacgtctgt ttactggtgt agtaccgatc ttagtggaat tagacggcga
     2761 cgtgaacggt cataaattta gcgtgcgcgg cgagggcgaa ggtgacgcta ccaatggtaa
     2821 attgaccctg aagtttattt gcacaacagg caaattaccc gttccgtggc ccaccttagt
     2881 gaccaccctg acctatggcg ttcagtgctt cagtcgttac cctaaacata tgaaacaaca
     2941 cgattttttc aaatcagcca tgcctgaagg atatgttcaa gagcgtacaa tcagcttcaa
     3001 gaaggatggc acctataaaa cgcgtgcgga agtgaaattt gaaggccgca cattagtaaa
     3061 tcgtatcgaa ctgaaaggtc gtgacttcaa agaagacggc aacattttag gccataaact
     3121 ggaatataac tttaattctc ataacgtgta tattacggcc gataaacaga agaatggtat
     3181 caaggcaaat ttcaaaattc gccataacgt ggaggacggc agcgttcaat tagcggatca
     3241 ttatcaacaa aacacgccga ttggtgatgg gcctgtactg ttacctcgca accactacct
     3301 gagcacccaa tctgccctga gcaaagatcc gaaagaaaaa cgcgatcaca tggttctgtt
     3361 agaattcgtg accgctgcag gcattacgca cggaatggac gaacgctaca agggcggtag
     3421 caaaaagcgt aagaagcgtt aataactcga gtctggtaaa gaaaccgctg ctgcgaaatt
     3481 tgaacgccag cacatggact cgtctactag cgcagcttaa ttaacctagg ctgctgccac
     3541 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     3601 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
     3661 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3721 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3781 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3841 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3901 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3961 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4021 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4081 acgtttacaa tttctggcgg cacgatggca tgagattatc aaaaaggatc ttcacctaga
     4141 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4201 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4261 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4321 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4381 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4441 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4501 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4561 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4621 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4681 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4741 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4801 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4861 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4921 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4981 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5041 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatcatga ttgaagcatt
     5101 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5161 ataggtcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5221 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5281 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5341 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     5401 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5461 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5521 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5581 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5641 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5701 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5761 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5821 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5881 tgctcacatg ttctttcctg cgttatcccc t
//
