LOCUS       sequence-405658-        5959 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 5959)
  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 5959)
  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)
FEATURES             Location/Qualifiers
     source          1..5959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1263..1454
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1448..1470)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1556..1695
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1611..1630
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1710..1727)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1881..2469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1961..1980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2550..2642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2643..2711,2712..3503)
                     /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(2861..2880)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3618..4073)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3755..3776)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3967..3986
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4110..4157)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4153..4171
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4209..4253)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(4318..4335)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(4352..5059)
                     /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(4630..4649)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4899..4917
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(5066..5083)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             5097..5119
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5134..5158
                     /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(5158..5177)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5159..5177)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5265..5284)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5490..5567
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5568..5959,1..691)
                     /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(5587..5606)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
       61 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
      121 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
      181 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
      241 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
      301 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
      361 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggaca tctcggtagt
      421 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
      481 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
      541 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
      601 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
      661 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca
      721 ctcattaggc accgggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
      781 tccggtgggc gcggggcatg actagcatga tcgtgctcct gtcgttgagg acccggctag
      841 gctggcgggg ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa
      901 gcgactgctg ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc
      961 gtgtttcgta aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct
     1021 gcatcgcagg atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct
     1081 ggcattgacc ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac
     1141 cctcacaacg ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc
     1201 tctctcgttt catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat
     1261 cagtgaccaa acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat
     1321 taacgcttct ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat
     1381 cgcttcacga ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg
     1441 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     1501 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag
     1561 ccatgaccca gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga
     1621 gcagattgta ctgagagtgc accattgcgg tgtgaaatac cgcacagatg cgtaaggaga
     1681 aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     1741 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     1801 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     1861 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     1921 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     1981 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2041 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2101 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2161 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2221 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2281 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     2341 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2401 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2461 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     2521 tcacgttaag ggattttggt catgacctat ttgtttattt ttctaaatac attcaaatat
     2581 gtatccgctc atgagacaat aaccctgata aatgcttcaa tcatgattga aaaaggaaga
     2641 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     2701 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     2761 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2821 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2881 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2941 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     3001 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     3061 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     3121 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     3181 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     3241 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     3301 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     3361 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     3421 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     3481 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     3541 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     3601 tgccatcgtg ccgccagaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt
     3661 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3721 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3781 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3841 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3901 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3961 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     4021 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc
     4081 gccaatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc
     4141 ccaaggggtt atgctagtta ttgctcagcg gtggcagcag cctaggttaa ttaagctgcg
     4201 ctagtagacg agtccatgtg ctggcgttca aatttcgcag cagcggtttc tttaccagac
     4261 tcgagtgcgg ccgcaagctt gtcgacggag ctcgaattcg gatccgatat cagccatgga
     4321 accgcgtggc accagggtac ccagatcttc attacttgta cagctcgtcc atgccgccgg
     4381 tggagtggcg gccctcggcg cgttcgtact gttccacgat ggtgtagtcc tcgttgtggg
     4441 aggtgatgtc caacttgatg ttgacgttgt aggcgccggg cagctgcacg ggcttcttgg
     4501 ccttgtaggt ggtcttgacc tcagcgtcgt agtggccgcc gtccttcagc ttcagcctct
     4561 gcttgatctc gcccttcagg gcgccgtcct cggggtacat ccgctcggag gaggcctccc
     4621 agcccatggt cttcttctgc attacggggc cgtcggaggg gaagttggtg ccgcgcagct
     4681 tcaccttgta gatgaactcg ccgtcctgga gggaggagtc ctgggtcacg gtcaccacgc
     4741 cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttca
     4801 agtagtcggg gatgtcggcg gggtgcttca cgtaggcctt ggagccgtac atgaactgag
     4861 gggacaggat gtcccaggcg aagggcaggg ggccaccctt ggtcaccttc agcttggcgg
     4921 tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt
     4981 tcacggagcc ctccatgtgc accttgaagc gcatgaactc cttgatgatg gccatgttat
     5041 cctcctcgcc cttgctcacg ccaccgtgat ggtggtggtg atgacccata tgtatatctc
     5101 cttcttaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc
     5161 tatagtgagt cgtattaatt tcgcgggatc gagatcgatc tcgatcctct acgccggacg
     5221 catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat
     5281 caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg
     5341 tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt
     5401 ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga
     5461 gtcgcataag ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg
     5521 gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa
     5581 cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga
     5641 accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc
     5701 tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg
     5761 gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat
     5821 ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg
     5881 aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta
     5941 actatccgct ggatgacca
//
