LOCUS       sequence-405657-        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(+6)-H3
FEATURES             Location/Qualifiers
     source          1..5938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             891..1082
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1076..1098)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1184..1323
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1239..1258
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1338..1355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1509..2097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1589..1608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2178..2270
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2271..2339,2340..3131)
                     /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(2489..2508)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3246..3701)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3383..3404)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3595..3614
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3738..3785)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3781..3799
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3837..3881)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(4674..4691)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4704..4726
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4741..4765
                     /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(4765..4784)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4766..4784)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4872..4891)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5097..5174
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5175..5938,1..319)
                     /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(5194..5213)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cgccattacc gagtccgggc tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga
       61 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
      121 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
      181 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
      241 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
      301 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
      361 cgggatctcg accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc
      421 ggggcatgac tagcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt
      481 gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct
      541 gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa
      601 gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat
      661 gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct
      721 gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt
      781 ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca
      841 tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac
      901 aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg
      961 agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc
     1021 acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     1081 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     1141 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     1201 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     1261 gagagtgcac cattgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     1321 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1381 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1441 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1501 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1561 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1621 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1681 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1741 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1801 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1861 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1921 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     1981 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2041 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2101 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2161 attttggtca tgacctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2221 gagacaataa ccctgataaa tgcttcaatc atgattgaaa aaggaagagt atgagtattc
     2281 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2341 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2401 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2461 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2521 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2581 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2641 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2701 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2761 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2821 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2881 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2941 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3001 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3061 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3121 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3181 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg ccatcgtgcc
     3241 gccagaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3301 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3361 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3421 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3481 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3541 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3601 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3661 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc caatccggat
     3721 atagttcctc ctttcagcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     3781 gctagttatt gctcagcggt ggcagcagcc taggttaatt aagctgcgct agtagacgag
     3841 tccatgtgct ggcgttcaaa tttcgcagca gcggtttctt taccagactc gagttattac
     3901 tttttacgtc ttcgtttttt cttgcgtcta cgctttttct tgcgtctacg cttcttgtag
     3961 cgttcgtcca ttccgtgcgt aatgcctgca gcggtcacga attctaacag aaccatgtga
     4021 tcgcgttttt ctttcggatc tttgctcagg gcagattggg tgctcaggta gtggttgcga
     4081 ggtaacagta caggcccatc accaatcggc gtgttttgtt gataatgatc cgctaattga
     4141 acgctgccgt cctccacgtt atggcgaatt ttgaaatttg ccttgatacc attcttctgt
     4201 ttatcggccg taatatacac gttatgagaa ttaaagttat attccagttt atggcctaaa
     4261 atgttgccgt cttctttgaa gtcacgacct ttcagttcga tacgatttac taatgtgcgg
     4321 ccttcaaatt tcacttccgc acgcgtttta taggtgccat ccttcttgaa gctgattgta
     4381 cgctcttgaa catatccttc aggcatggct gatttgaaaa aatcgtgttg tttcatatgt
     4441 ttagggtaac gactgaagca ctgaacgcca taggtcaggg tggtcactaa ggtgggccac
     4501 ggaacgggta atttgcctgt tgtgcaaata aacttcaggg tcaatttacc attggtagcg
     4561 tcaccttcgc cctcgccgcg cacgctaaat ttatgaccgt tcacgtcgcc gtctaattcc
     4621 actaagatcg gtactacacc agtaaacaga cgttcacctt tgctagcgcc accgtgatgg
     4681 tggtggtgat gacccatggt atatctcctt cttaaagtta aacaaaatta tttctagagg
     4741 ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag
     4801 atcgatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     4861 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     4921 gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     4981 gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg gcctcaacct
     5041 actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag atcccggaca
     5101 ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     5161 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     5221 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     5281 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     5341 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     5401 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     5461 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     5521 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     5581 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     5641 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     5701 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     5761 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     5821 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     5881 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcg
//
