LOCUS       sequence-382882-        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-4 GFP(0)-tag1
FEATURES             Location/Qualifiers
     source          1..5911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     158..180
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(174..365)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     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     2410..2429
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2410..2428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2429..2453
                     /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             2468..2490
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2503..2520
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             3286..3330
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3368..3386)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3382..3429
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3466..3921
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3553..3572)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3763..3784
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(4036..4827,4828..4896))
                     /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     4659..4678
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4897..4989)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5070..5658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5559..5578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5812..5829
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5844..5911,1..72)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(join(5909..5911,1..17))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
       61 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
      121 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
      181 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
      241 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
      301 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
      361 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
      421 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
      481 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
      541 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
      601 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
      661 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
      721 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
      781 cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat 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 accgagatgt 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 gatctcgatc
     2401 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     2461 aaataatttt gtttaacttt aagaaggaga tataccatgg gtcatcacca ccaccatcac
     2521 ggtggcgcta gcaaaggtga acgtctgttt actggtgtag taccgatctt agtggaatta
     2581 gacggcgacg tgaacggtca taaatttagc gtgcgcggcg agggcgaagg tgacgctacc
     2641 aatggtaaat tgaccctgaa gtttatttgc acaacaggca aattacccgt tccgtggccc
     2701 accttagtga ccaccctgac ctatggcgtt cagtgcttca gtcgttaccc tgatcatatg
     2761 aaacaacacg attttttcaa atcagccatg cctgaaggat atgttcaaga gcgtacaatc
     2821 agcttcaagg acgatggcac ctataaaacg cgtgcggaag tgaaatttga aggcgacaca
     2881 ttagtaaatc gtatcgaact gaaaggtcgt gacttcaaag aagacggcaa cattttaggc
     2941 cataaactgg aatataactt taattctcat aacgtgtata ttacggccga taaacagaag
     3001 aatggtatca aggcaaattt caaaattcgc cataacgtgg aggacggcag cgttcaatta
     3061 gcggatcatt atcaacaaaa cacgccgatt ggtgatgggc ctgtactgtt acctcgcaac
     3121 cactacctga gcacccaatc tgccctgagc aaagatccga aagaaaaacg cgatcacatg
     3181 gttctgttag aattcgtgac cgctgcaggc attacgcacg gaatggacga acgctacaag
     3241 ggcggtagca aaaagcgtaa gaagcgttaa taactcgagt ctggtaaaga aaccgctgct
     3301 gcgaaatttg aacgccagca catggactcg tctactagcg cagcttaatt aacctaggct
     3361 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     3421 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg
     3481 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3541 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3601 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     3661 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3721 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3781 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     3841 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3901 aaatattaac gtttacaatt tctggcggca cgatggcatg agattatcaa aaaggatctt
     3961 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4021 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4081 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4141 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4201 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4261 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4321 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4381 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4441 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     4501 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     4561 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     4621 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     4681 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     4741 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     4801 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     4861 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatcatgatt
     4921 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4981 ataaacaaat aggtcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5041 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5101 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5161 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     5221 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5281 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5341 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5401 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5461 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     5521 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     5581 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     5641 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     5701 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5761 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5821 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5881 atttcacacc gcaatggtgc actctcagta c
//
