LOCUS       sequence-405639-        5968 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 5968)
  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 5968)
  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)-L3
FEATURES             Location/Qualifiers
     source          1..5968
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1115..1306
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1300..1322)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1408..1547
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1463..1482
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1562..1579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1733..2321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1813..1832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2402..2494
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2495..2563,2564..3355)
                     /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(2713..2732)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3470..3925)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3607..3628)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3819..3838
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3962..4009)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4005..4023
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4061..4105)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(4928..4945)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4958..4980
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4995..5019
                     /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(5019..5038)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5020..5038)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5126..5145)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5351..5428
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5429..5968,1..543)
                     /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(5448..5467)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 caaatcgcgc tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc
       61 tggcataaat atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg
      121 agtgccatgt ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact
      181 gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc
      241 gggctgcgcg ttggtgcgga catctcggta gtgggatacg acgataccga agacagctca
      301 tgttatatcc cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc
      361 gtggaccgct tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc
      421 gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata cgcaaaccgc ctctccccgc
      481 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
      541 tgagcgcaac gcaattaatg taagttagct cactcattag gcaccgggat ctcgaccgat
      601 gcccttgaga gccttcaacc cagtcagctc cttccggtgg gcgcggggca tgactagcat
      661 gatcgtgctc ctgtcgttga ggacccggct aggctggcgg ggttgcctta ctggttagca
      721 gaatgaatca ccgatacgcg agcgaacgtg aagcgactgc tgctgcaaaa cgtctgcgac
      781 ctgagcaaca acatgaatgg tcttcggttt ccgtgtttcg taaagtctgg aaacgcggaa
      841 gtcagcgccc tgcaccatta tgttccggat ctgcatcgca ggatgctgct ggctaccctg
      901 tggaacacct acatctgtat taacgaagcg ctggcattga ccctgagtga tttttctctg
      961 gtcccgccgc atccataccg ccagttgttt accctcacaa cgttccagta accgggcatg
     1021 ttcatcatca gtaacccgta tcgtgagcat cctctctcgt ttcatcggta tcattacccc
     1081 catgaacaga aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct
     1141 taacatggcc cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct
     1201 ggacgcggat gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta
     1261 ccgcagctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     1321 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     1381 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg
     1441 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccattgc
     1501 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     1561 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1621 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1681 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1741 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1801 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1861 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1921 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1981 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2041 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2101 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2161 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2221 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2281 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2341 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgacct
     2401 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     2461 taaatgcttc aatcatgatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     2521 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     2581 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     2641 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     2701 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     2761 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2821 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2881 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2941 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     3001 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     3061 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     3121 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     3181 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     3241 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     3301 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     3361 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     3421 gatctaggtg aagatccttt ttgataatct catgccatcg tgccgccaga aattgtaaac
     3481 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3541 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3601 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3661 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3721 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3781 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3841 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3901 cttaatgcgc cgctacaggg cgcgtcccat tcgccaatcc ggatatagtt cctcctttca
     3961 gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag
     4021 cggtggcagc agcctaggtt aattaagctg cgctagtaga cgagtccatg tgctggcgtt
     4081 caaatttcgc agcagcggtt tctttaccag actcgagtta ttacttccgc gatttcttcc
     4141 ctcttcgacc cttccgcgat ttcttccctc ttcgaccctt tcgagatttc ttgcctcttc
     4201 gtcctttctt gtacagctcg tccattccgt gagtaatgcc tgcagcggtc acgaattcta
     4261 acagaaccat gtgatcgcgt ttttcgttcg gatctttgct cagtgctgac tgggtgctca
     4321 ggtagtggtt gtcaggtaac agtacaggcc cgtcaccaat cggcgtgttt tgttgataat
     4381 gatccgctaa ttgaacgctg ccgtcttcca cgttatggcg aattttgaaa tttgccttga
     4441 taccgttttt ctgtttatcg gccgtaatat acacgttatg agaattaaag ttatattcca
     4501 gcttatggcc taaaatgttg ccgtcttctt tgaagtcgat acctttcagt tcgatacggt
     4561 ttactaatgt gtcgccttca aatttcactt ccgcacgcgt tttataggtg ccatcgtcct
     4621 tgaagctgat tgtacgctct tgaacatatc cttcaggcat ggctgatttg aaaaaatcgt
     4681 gttgtttcat atgatctggg taacgactga agcactgaac gccataggtc agggtggtca
     4741 ctaaggtggg ccacggaacg ggtaatttgc ctgttgtgca aataaacttc agggtcaatt
     4801 taccattggt agcgtcacct tcgccttcgc cgcgcacgct aaatttgtga ccgttcacgt
     4861 cgccgtctaa ttccactaag atcggtacta cacccgtaaa cagctcttca cctttgctag
     4921 cgccaccgtg atggtggtgg tgatgaccca tggtatatct ccttcttaaa gttaaacaaa
     4981 attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag tcgtattaat
     5041 ttcgcgggat cgagatcgat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc
     5101 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg
     5161 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg
     5221 gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc
     5281 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt
     5341 cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
     5401 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     5461 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     5521 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     5581 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     5641 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     5701 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     5761 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     5821 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     5881 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     5941 ggagcatctg gtcgcattgg gtcaccag
//
