LOCUS       sequence-405661-        5930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent protein with charge-patterned cationic peptide to 
            promote biomolecular condensate formation in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5930)
  AUTHORS   Liao J, Yeong V, Obermeyer AC
  TITLE     Charge-Patterned Disordered Peptides Tune Intracellular Phase 
            Separation in Bacteria.
  JOURNAL   ACS Synth Biol. 2024 Feb 3. doi: 10.1021/acssynbio.3c00564.
  PUBMED    38308651
REFERENCE   2  (bases 1 to 5930)
  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: "ACS Synth 
            Biol. 2024 Feb 3. doi: 10.1021/acssynbio.3c00564."
COMMENT     Sequence Label: pET-6xHis-mCherry(-6)-L2
FEATURES             Location/Qualifiers
     source          1..5930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1058..1249
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1243..1265)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1351..1490
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1406..1425
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1505..1522)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1676..2264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1756..1775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2345..2437
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2438..2506,2507..3298)
                     /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     complement(2656..2675)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3413..3868)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3550..3571)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3762..3781
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3905..3952)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3948..3966
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4004..4048)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(4121..4825)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(4396..4415)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4665..4683
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(4832..4849)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4863..4885
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4900..4924
                     /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)."
     primer_bind     complement(4924..4943)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4925..4943)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5031..5050)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5256..5333
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5334..5930,1..486)
                     /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     complement(5353..5372)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
       61 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
      121 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
      181 tccgggctgc gcgttggtgc ggacatctcg gtagtgggat acgacgatac cgaagacagc
      241 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
      301 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
      361 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
      421 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
      481 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcaccgg gatctcgacc
      541 gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg gcatgactag
      601 catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg cggggttgcc ttactggtta
      661 gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc
      721 gacctgagca acaacatgaa tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg
      781 gaagtcagcg ccctgcacca ttatgttccg gatctgcatc gcaggatgct gctggctacc
      841 ctgtggaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag tgatttttct
      901 ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca gtaaccgggc
      961 atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg gtatcattac
     1021 ccccatgaac agaaatcccc cttacacgga ggcatcagtg accaaacagg aaaaaaccgc
     1081 ccttaacatg gcccgcttta tcagaagcca gacattaacg cttctggaga aactcaacga
     1141 gctggacgcg gatgaacagg cagacatctg tgaatcgctt cacgaccacg ctgatgagct
     1201 ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     1261 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     1321 cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag
     1381 cggagtgtat actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     1441 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgctcttccg
     1501 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     1561 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     1621 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1681 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1741 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1801 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1861 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1921 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1981 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2041 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2101 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2161 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2221 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2281 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2341 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2401 tgataaatgc ttcaatcatg attgaaaaag gaagagtatg agtattcaac atttccgtgt
     2461 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2521 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     2581 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     2641 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     2701 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     2761 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     2821 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     2881 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     2941 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3001 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3061 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3121 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3181 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3241 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3301 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3361 aaggatctag gtgaagatcc tttttgataa tctcatgcca tcgtgccgcc agaaattgta
     3421 aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     3481 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     3541 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     3601 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     3661 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     3721 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     3781 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     3841 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccaa tccggatata gttcctcctt
     3901 tcagcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct agttattgct
     3961 cagcggtggc agcagcctag gttaattaag ctgcgctagt agacgagtcc atgtgctggc
     4021 gttcaaattt cgcagcagcg gtttctttac cagactcgag tcattacttt cgagatttct
     4081 tgcctcttcg tccctttcga gatttcttgc ctcttcgtcc cttgtacagc tcgtccatgc
     4141 cgccggtgga gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt
     4201 tgtgggaggt gatgtccaac ttgatgttga cgttgtaggc gccgggcagc tgcacgggct
     4261 tcttggcctt gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca
     4321 gcctctgctt gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg
     4381 cctcccagcc catggtcttc ttctgcatta cggggccgtc ggaggggaag ttggtgccgc
     4441 gcagcttcac cttgtagatg aactcgccgt cctggaggga ggagtcctgg gtcacggtca
     4501 ccacgccgcc gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca
     4561 gcttcaagta gtcggggatg tcggcggggt gcttcacgta ggccttggag ccgtacatga
     4621 actgagggga caggatgtcc caggcgaagg gcagggggcc acccttggtc accttcagct
     4681 tggcggtctg ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt
     4741 ggccgttcac ggagccctcc atgtgcacct tgaagcgcat gaactccttg atgatggcca
     4801 tgttatcctc ctcgcccttg ctcacgccac cgtgatggtg gtggtgatga cccatatgta
     4861 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa
     4921 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tcgatctcga tcctctacgc
     4981 cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc
     5041 cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg
     5101 cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc
     5161 accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat
     5221 gcaggagtcg cataagggag agcgtcgaga tcccggacac catcgaatgg cgcaaaacct
     5281 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     5341 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     5401 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     5461 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     5521 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     5581 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     5641 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc agtgggctga
     5701 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     5761 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     5821 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     5881 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
//
