LOCUS       sequence-405656-        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(+6)-H2
FEATURES             Location/Qualifiers
     source          1..5920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1049..1240
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1234..1256)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1342..1481
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1397..1416
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1496..1513)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1667..2255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1747..1766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2336..2428
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2429..2497,2498..3289)
                     /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(2647..2666)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3404..3859)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3541..3562)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3753..3772
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3896..3943)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3939..3957
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3995..4039)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(4814..4831)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4844..4866
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4881..4905
                     /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(4905..4924)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4906..4924)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5012..5031)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5237..5314
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5315..5920,1..477)
                     /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(5334..5353)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
       61 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
      121 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
      181 cgcgttggtg cggacatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
      241 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
      301 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
      361 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
      421 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
      481 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt
      541 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta gcatgatcgt
      601 gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga
      661 atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc
      721 aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc
      781 gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac
      841 acctacatct gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg
      901 ccgcatccat accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc
      961 atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa
     1021 cagaaatccc ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat
     1081 ggcccgcttt atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc
     1141 ggatgaacag gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag
     1201 ctgcctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     1261 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     1321 gggtgttggc gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta
     1381 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca ttgcggtgtg
     1441 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
     1501 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2101 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2221 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg acctatttgt
     2341 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2401 cttcaatcat gattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2461 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2521 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2581 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     2641 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     2701 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2761 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2821 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     2881 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2941 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3001 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3061 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3121 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3181 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3241 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3301 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3361 ggtgaagatc ctttttgata atctcatgcc atcgtgccgc cagaaattgt aaacgttaat
     3421 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3481 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3541 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3601 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3661 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3721 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3781 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3841 gcgccgctac agggcgcgtc ccattcgcca atccggatat agttcctcct ttcagcaaaa
     3901 aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc tcagcggtgg
     3961 cagcagccta ggttaattaa gctgcgctag tagacgagtc catgtgctgg cgttcaaatt
     4021 tcgcagcagc ggtttcttta ccagactcga gttattattt cttgcgtcta cgctttttct
     4081 tgcgtctacg cttcttgtag cgttcgtcca ttccgtgcgt aatgcctgca gcggtcacga
     4141 attctaacag aaccatgtga tcgcgttttt ctttcggatc tttgctcagg gcagattggg
     4201 tgctcaggta gtggttgcga ggtaacagta caggcccatc accaatcggc gtgttttgtt
     4261 gataatgatc cgctaattga acgctgccgt cctccacgtt atggcgaatt ttgaaatttg
     4321 ccttgatacc attcttctgt ttatcggccg taatatacac gttatgagaa ttaaagttat
     4381 attccagttt atggcctaaa atgttgccgt cttctttgaa gtcacgacct ttcagttcga
     4441 tacgatttac taatgtgcgg ccttcaaatt tcacttccgc acgcgtttta taggtgccat
     4501 ccttcttgaa gctgattgta cgctcttgaa catatccttc aggcatggct gatttgaaaa
     4561 aatcgtgttg tttcatatgt ttagggtaac gactgaagca ctgaacgcca taggtcaggg
     4621 tggtcactaa ggtgggccac ggaacgggta atttgcctgt tgtgcaaata aacttcaggg
     4681 tcaatttacc attggtagcg tcaccttcgc cctcgccgcg cacgctaaat ttatgaccgt
     4741 tcacgtcgcc gtctaattcc actaagatcg gtactacacc agtaaacaga cgttcacctt
     4801 tgctagcgcc accgtgatgg tggtggtgat gacccatggt atatctcctt cttaaagtta
     4861 aacaaaatta tttctagagg ggaattgtta tccgctcaca attcccctat agtgagtcgt
     4921 attaatttcg cgggatcgag atcgatctcg atcctctacg ccggacgcat cgtggccggc
     4981 atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa
     5041 gatcgggctc gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc
     5101 cccgtggccg ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg
     5161 gtgctcaacg gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga
     5221 gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata
     5281 gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt
     5341 cgcagagtat gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca
     5401 cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc
     5461 caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc
     5521 cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca
     5581 actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc
     5641 ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga
     5701 tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga
     5761 tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact
     5821 gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt
     5881 aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
//
