LOCUS       sequence-405650-        5920 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 5920)
  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 5920)
  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-GFP(0)-H2
FEATURES             Location/Qualifiers
     source          1..5920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1052..1243
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1237..1259)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1345..1484
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1400..1419
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1499..1516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1670..2258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1750..1769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2339..2431
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2432..2500,2501..3292)
                     /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(2650..2669)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3407..3862)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3544..3565)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3756..3775
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3899..3946)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3942..3960
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3998..4042)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(4817..4834)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4847..4869
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4884..4908
                     /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(4908..4927)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4909..4927)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5015..5034)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5240..5317
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5318..5920,1..480)
                     /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(5337..5356)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg cgactggagt
       61 gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt tcccactgcg
      121 atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac cgagtccggg
      181 ctgcgcgttg gtgcggacat ctcggtagtg ggatacgacg ataccgaaga cagctcatgt
      241 tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca aaccagcgtg
      301 gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct gttgcccgtc
      361 tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg
      421 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
      481 gcgcaacgca attaatgtaa gttagctcac tcattaggca ccgggatctc gaccgatgcc
      541 cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga ctagcatgat
      601 cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg gttagcagaa
      661 tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg
      721 agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc
      781 agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc taccctgtgg
      841 aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc
      901 ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc
      961 atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttacccccat
     1021 gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa
     1081 catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca acgagctgga
     1141 cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg agctttaccg
     1201 cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     1261 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     1321 agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt
     1381 gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccattgcggt
     1441 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct
     1501 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1561 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1621 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1681 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1741 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1801 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1861 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1921 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1981 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2041 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2101 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2161 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2221 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2281 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgacctatt
     2341 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2401 atgcttcaat catgattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2461 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2521 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2581 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2641 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2701 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2761 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2821 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2881 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2941 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3001 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3061 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3121 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3181 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3241 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     3301 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     3361 ctaggtgaag atcctttttg ataatctcat gccatcgtgc cgccagaaat tgtaaacgtt
     3421 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3481 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3541 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3601 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3661 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3721 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3781 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3841 aatgcgccgc tacagggcgc gtcccattcg ccaatccgga tatagttcct cctttcagca
     3901 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat tgctcagcgg
     3961 tggcagcagc ctaggttaat taagctgcgc tagtagacga gtccatgtgc tggcgttcaa
     4021 atttcgcagc agcggtttct ttaccagact cgagttatta tttcttgcgt ctacgctttt
     4081 tcttgcgtct acgcttcttg tagcgttcgt ccattccgtg cgtaatgcct gcagcggtca
     4141 cgaattctaa cagaaccatg tgatcgcgtt tttctttcgg atctttgctc agggcagatt
     4201 gggtgctcag gtagtggttg cgaggtaaca gtacaggccc atcaccaatc ggcgtgtttt
     4261 gttgataatg atccgctaat tgaacgctgc cgtcctccac gttatggcga attttgaaat
     4321 ttgccttgat accattcttc tgtttatcgg ccgtaatata cacgttatga gaattaaagt
     4381 tatattccag tttatggcct aaaatgttgc cgtcttcttt gaagtcacga cctttcagtt
     4441 cgatacgatt tactaatgtg tcgccttcaa atttcacttc cgcacgcgtt ttataggtgc
     4501 catcgtcctt gaagctgatt gtacgctctt gaacatatcc ttcaggcatg gctgatttga
     4561 aaaaatcgtg ttgtttcata tgatcagggt aacgactgaa gcactgaacg ccataggtca
     4621 gggtggtcac taaggtgggc cacggaacgg gtaatttgcc tgttgtgcaa ataaacttca
     4681 gggtcaattt accattggta gcgtcacctt cgccctcgcc gcgcacgcta aatttatgac
     4741 cgttcacgtc gccgtctaat tccactaaga tcggtactac accagtaaac agacgttcac
     4801 ctttgctagc gccaccgtga tggtggtggt gatgacccat ggtatatctc cttcttaaag
     4861 ttaaacaaaa ttatttctag aggggaattg ttatccgctc acaattcccc tatagtgagt
     4921 cgtattaatt tcgcgggatc gagatcgatc tcgatcctct acgccggacg catcgtggcc
     4981 ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat caccgatggg
     5041 gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg tatggtggca
     5101 ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt ccttgcggcg
     5161 gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga gtcgcataag
     5221 ggagagcgtc gagatcccgg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
     5281 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     5341 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     5401 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     5461 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     5521 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     5581 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     5641 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     5701 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     5761 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     5821 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     5881 attaagttct gtctcggcgc gtctgcgtct ggctggctgg
//
