LOCUS       sequence-405651-        5938 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 5938)
  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 5938)
  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)-H3
FEATURES             Location/Qualifiers
     source          1..5938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1054..1245
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1239..1261)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1347..1486
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1402..1421
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1501..1518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1672..2260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1752..1771)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2341..2433
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2434..2502,2503..3294)
                     /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(2652..2671)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3409..3864)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3546..3567)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3758..3777
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3901..3948)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3944..3962
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4000..4044)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(4837..4854)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4867..4889
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4904..4928
                     /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(4928..4947)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4929..4947)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5035..5054)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5260..5337
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5338..5938,1..482)
                     /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(5357..5376)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga
       61 gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg
      121 cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg
      181 ggctgcgcgt tggtgcggac atctcggtag tgggatacga cgataccgaa gacagctcat
      241 gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg
      301 tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg
      361 tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg
      421 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt
      481 gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc tcgaccgatg
      541 cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactagcatg
      601 atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag
      661 aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc
      721 tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag
      781 tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt
      841 ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg
      901 tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt
      961 tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc
     1021 atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt
     1081 aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg
     1141 gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac
     1201 cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     1261 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     1321 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     1381 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccattgcg
     1441 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc
     1501 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1561 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     1621 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     1681 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1741 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1801 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1861 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1921 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1981 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2041 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2101 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2161 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2221 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2281 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaccta
     2341 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2401 aaatgcttca atcatgattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     2461 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     2521 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2581 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2641 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2701 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2761 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     2821 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2881 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2941 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3001 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3061 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3121 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3181 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3241 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     3301 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     3361 atctaggtga agatcctttt tgataatctc atgccatcgt gccgccagaa attgtaaacg
     3421 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3481 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3541 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3601 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3661 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3721 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3781 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3841 ttaatgcgcc gctacagggc gcgtcccatt cgccaatccg gatatagttc ctcctttcag
     3901 caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctagtt attgctcagc
     3961 ggtggcagca gcctaggtta attaagctgc gctagtagac gagtccatgt gctggcgttc
     4021 aaatttcgca gcagcggttt ctttaccaga ctcgagttat tactttttac gtcttcgttt
     4081 tttcttgcgt ctacgctttt tcttgcgtct acgcttcttg tagcgttcgt ccattccgtg
     4141 cgtaatgcct gcagcggtca cgaattctaa cagaaccatg tgatcgcgtt tttctttcgg
     4201 atctttgctc agggcagatt gggtgctcag gtagtggttg cgaggtaaca gtacaggccc
     4261 atcaccaatc ggcgtgtttt gttgataatg atccgctaat tgaacgctgc cgtcctccac
     4321 gttatggcga attttgaaat ttgccttgat accattcttc tgtttatcgg ccgtaatata
     4381 cacgttatga gaattaaagt tatattccag tttatggcct aaaatgttgc cgtcttcttt
     4441 gaagtcacga cctttcagtt cgatacgatt tactaatgtg tcgccttcaa atttcacttc
     4501 cgcacgcgtt ttataggtgc catcgtcctt gaagctgatt gtacgctctt gaacatatcc
     4561 ttcaggcatg gctgatttga aaaaatcgtg ttgtttcata tgatcagggt aacgactgaa
     4621 gcactgaacg ccataggtca gggtggtcac taaggtgggc cacggaacgg gtaatttgcc
     4681 tgttgtgcaa ataaacttca gggtcaattt accattggta gcgtcacctt cgccctcgcc
     4741 gcgcacgcta aatttatgac cgttcacgtc gccgtctaat tccactaaga tcggtactac
     4801 accagtaaac agacgttcac ctttgctagc gccaccgtga tggtggtggt gatgacccat
     4861 ggtatatctc cttcttaaag ttaaacaaaa ttatttctag aggggaattg ttatccgctc
     4921 acaattcccc tatagtgagt cgtattaatt tcgcgggatc gagatcgatc tcgatcctct
     4981 acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata
     5041 tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt
     5101 tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc
     5161 atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc
     5221 taatgcagga gtcgcataag ggagagcgtc gagatcccgg acaccatcga atggcgcaaa
     5281 acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg
     5341 aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc
     5401 cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg
     5461 atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg caaacagtcg
     5521 ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg
     5581 gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga
     5641 agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg
     5701 ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc tgcctgcact
     5761 aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag tattattttc
     5821 tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg tcaccagcaa
     5881 atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct ggctggct
//
