LOCUS       sequence-405642-        5941 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 5941)
  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 5941)
  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..5941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1053..1244
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1238..1260)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1346..1485
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1401..1420
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1500..1517)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1671..2259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1751..1770)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2340..2432
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2433..2501,2502..3293)
                     /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(2651..2670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3408..3863)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3545..3566)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3757..3776
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3900..3947)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3943..3961
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3999..4043)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(4839..4856)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4869..4891
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4906..4930
                     /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(4930..4949)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4931..4949)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5037..5056)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5262..5339
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5340..5941,1..481)
                     /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(5359..5378)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcataaatat ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag
       61 tgccatgtcc ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc
      121 gatgctggtt gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg
      181 gctgcgcgtt ggtgcggaca tctcggtagt gggatacgac gataccgaag acagctcatg
      241 ttatatcccg ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt
      301 ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt
      361 ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc
      421 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
      481 agcgcaacgc aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc
      541 ccttgagagc cttcaaccca gtcagctcct tccggtgggc gcggggcatg actagcatga
      601 tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact ggttagcaga
      661 atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg tctgcgacct
      721 gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa acgcggaagt
      781 cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg
      841 gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt tttctctggt
      901 cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac cgggcatgtt
      961 catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc attaccccca
     1021 tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa accgccctta
     1081 acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc aacgagctgg
     1141 acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat gagctttacc
     1201 gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     1261 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     1321 cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc gatagcggag
     1381 tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accattgcgg
     1441 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc
     1501 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1561 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1621 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1681 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1741 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1801 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1861 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1921 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1981 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2041 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2101 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2161 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2221 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2281 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgacctat
     2341 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2401 aatgcttcaa tcatgattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2461 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2521 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2581 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2641 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2701 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2761 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2821 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2881 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2941 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3001 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3061 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3121 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3181 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3241 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3301 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3361 tctaggtgaa gatccttttt gataatctca tgccatcgtg ccgccagaaa ttgtaaacgt
     3421 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3481 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3541 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3601 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3661 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3721 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3781 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3841 taatgcgccg ctacagggcg cgtcccattc gccaatccgg atatagttcc tcctttcagc
     3901 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg
     3961 gtggcagcag cctaggttaa ttaagctgcg ctagtagacg agtccatgtg ctggcgttca
     4021 aatttcgcag cagcggtttc tttaccagac tcgagttatt atttcttgcg tctacgcttt
     4081 ttcttgcgtc tacgcttttt cttccttctc ctctttttct tgtacagctc gtccattccg
     4141 tgagtaatgc ctgcagcggt cacgaattct aacagaacca tgtgatcgcg tttttcgttc
     4201 ggatctttgc tcagtgctga ctgggtgctc aggtagtggt tgtcaggtaa cagtacaggc
     4261 ccgtcaccaa tcggcgtgtt ttgttgataa tgatccgcta attgaacgct gccgtcttcc
     4321 acgttatggc gaattttgaa atttgccttg ataccgtttt tctgtttatc ggccgtaata
     4381 tacacgttat gagaattaaa gttatattcc agcttatggc ctaaaatgtt gccgtcttct
     4441 ttgaagtcga tacctttcag ttcgatacgg tttactaatg tgtcgccttc aaatttcact
     4501 tccgcacgcg ttttataggt gccatcgtcc ttgaagctga ttgtacgctc ttgaacatat
     4561 ccttcaggca tggctgattt gaaaaaatcg tgttgtttca tatgatctgg gtaacgactg
     4621 aagcactgaa cgccataggt cagggtggtc actaaggtgg gccacggaac gggtaatttg
     4681 cctgttgtgc aaataaactt cagggtcaat ttaccattgg tagcgtcacc ttcgccttcg
     4741 ccgcgcacgc taaatttgtg accgttcacg tcgccgtcta attccactaa gatcggtact
     4801 acacccgtaa acagctcttc acctttgcta gcgccaccgt gatggtggtg gtgatgaccc
     4861 atggtatatc tccttcttaa agttaaacaa aattatttct agaggggaat tgttatccgc
     4921 tcacaattcc cctatagtga gtcgtattaa tttcgcggga tcgagatcga tctcgatcct
     4981 ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta
     5041 tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg
     5101 tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt
     5161 gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt
     5221 cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc gaatggcgca
     5281 aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg
     5341 tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt
     5401 cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg
     5461 cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt
     5521 cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg
     5581 cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac
     5641 gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg
     5701 ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca
     5761 ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt
     5821 tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc
     5881 aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct
     5941 g
//
