LOCUS       sequence-405637-        5911 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 5911)
  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 5911)
  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)-L1
FEATURES             Location/Qualifiers
     source          1..5911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             889..1080
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1074..1096)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1182..1321
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1237..1256
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1336..1353)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1507..2095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1587..1606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2176..2268
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2269..2337,2338..3129)
                     /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(2487..2506)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3244..3699)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3381..3402)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3593..3612
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3736..3783)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3779..3797
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3835..3879)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(4645..4662)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4675..4697
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4712..4736
                     /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(4736..4755)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4737..4755)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4843..4862)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5068..5145
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5146..5911,1..317)
                     /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(5165..5184)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ccattaccga gtccgggctg cgcgttggtg cggacatctc ggtagtggga tacgacgata
       61 ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat tttcgcctgc
      121 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
      181 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
      241 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
      301 tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca ttaggcaccg
      361 ggatctcgac cgatgccctt gagagccttc aacccagtca gctccttccg gtgggcgcgg
      421 ggcatgacta gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc
      481 cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc
      541 aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt
      601 ctggaaacgc ggaagtcagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc
      661 tgctggctac cctgtggaac acctacatct gtattaacga agcgctggca ttgaccctga
      721 gtgatttttc tctggtcccg ccgcatccat accgccagtt gtttaccctc acaacgttcc
      781 agtaaccggg catgttcatc atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc
      841 ggtatcatta cccccatgaa cagaaatccc ccttacacgg aggcatcagt gaccaaacag
      901 gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag
      961 aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac
     1021 gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     1081 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     1141 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     1201 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
     1261 gagtgcacca ttgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     1321 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1381 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1441 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1501 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1561 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1621 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1681 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1741 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1801 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1861 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1921 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     1981 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     2041 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     2101 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2161 tttggtcatg acctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2221 gacaataacc ctgataaatg cttcaatcat gattgaaaaa ggaagagtat gagtattcaa
     2281 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2341 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2401 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2461 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2521 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2581 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2641 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2701 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2761 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2821 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2881 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2941 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3001 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3061 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3121 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3181 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgcc atcgtgccgc
     3241 cagaaattgt aaacgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3301 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3361 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3421 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3481 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3541 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3601 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3661 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca atccggatat
     3721 agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc
     3781 tagttattgc tcagcggtgg cagcagccta ggttaattaa gctgcgctag tagacgagtc
     3841 catgtgctgg cgttcaaatt tcgcagcagc ggtttcttta ccagactcga gttactttcg
     3901 agatttcttg cctcttcgtc ctttcttgta cagctcgtcc attccgtgag taatgcctgc
     3961 agcggtcacg aattctaaca gaaccatgtg atcgcgtttt tcgttcggat ctttgctcag
     4021 tgctgactgg gtgctcaggt agtggttgtc aggtaacagt acaggcccgt caccaatcgg
     4081 cgtgttttgt tgataatgat ccgctaattg aacgctgccg tcttccacgt tatggcgaat
     4141 tttgaaattt gccttgatac cgtttttctg tttatcggcc gtaatataca cgttatgaga
     4201 attaaagtta tattccagct tatggcctaa aatgttgccg tcttctttga agtcgatacc
     4261 tttcagttcg atacggttta ctaatgtgtc gccttcaaat ttcacttccg cacgcgtttt
     4321 ataggtgcca tcgtccttga agctgattgt acgctcttga acatatcctt caggcatggc
     4381 tgatttgaaa aaatcgtgtt gtttcatatg atctgggtaa cgactgaagc actgaacgcc
     4441 ataggtcagg gtggtcacta aggtgggcca cggaacgggt aatttgcctg ttgtgcaaat
     4501 aaacttcagg gtcaatttac cattggtagc gtcaccttcg ccttcgccgc gcacgctaaa
     4561 tttgtgaccg ttcacgtcgc cgtctaattc cactaagatc ggtactacac ccgtaaacag
     4621 ctcttcacct ttgctagcgc caccgtgatg gtggtggtga tgacccatgg tatatctcct
     4681 tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta
     4741 tagtgagtcg tattaatttc gcgggatcga gatcgatctc gatcctctac gccggacgca
     4801 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
     4861 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
     4921 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
     4981 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt
     5041 cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt
     5101 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     5161 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     5221 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     5281 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     5341 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     5401 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     5461 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     5521 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     5581 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     5641 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     5701 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     5761 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     5821 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     5881 aacgatcaga tggcgctggg cgcaatgcgc g
//
