LOCUS       sequence-405638-        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)-L2
FEATURES             Location/Qualifiers
     source          1..5941
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1013..1204
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1198..1220)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1306..1445
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1361..1380
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1460..1477)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1631..2219)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1711..1730)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2300..2392
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2393..2461,2462..3253)
                     /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(2611..2630)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3368..3823)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3505..3526)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3717..3736
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3860..3907)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3903..3921
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3959..4003)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(4799..4816)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4829..4851
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4866..4890
                     /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(4890..4909)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4891..4909)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4997..5016)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5222..5299
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5300..5941,1..441)
                     /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(5319..5338)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
       61 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
      121 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggaca tctcggtagt gggatacgac
      181 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
      241 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
      301 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
      361 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
      421 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
      481 accgggatct cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
      541 gcggggcatg actagcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg
      601 ttgccttact ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg
      661 ctgcaaaacg tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta
      721 aagtctggaa acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg
      781 atgctgctgg ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc
      841 ctgagtgatt tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg
      901 ttccagtaac cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt
      961 catcggtatc attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa
     1021 acaggaaaaa accgccctta acatggcccg ctttatcaga agccagacat taacgcttct
     1081 ggagaaactc aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga
     1141 ccacgctgat gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct
     1201 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     1261 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca
     1321 gtcacgtagc gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta
     1381 ctgagagtgc accattgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     1441 tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1501 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1561 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1621 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1681 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1741 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1801 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1861 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1921 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1981 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     2041 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     2101 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2161 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2221 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2281 ggattttggt catgacctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     2341 atgagacaat aaccctgata aatgcttcaa tcatgattga aaaaggaaga gtatgagtat
     2401 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     2461 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     2521 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     2581 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     2641 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     2701 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     2761 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     2821 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     2881 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     2941 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3001 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3061 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3121 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3181 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3241 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3301 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgccatcgtg
     3361 ccgccagaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3421 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3481 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3541 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3601 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3661 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3721 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3781 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccaatccgg
     3841 atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt
     3901 atgctagtta ttgctcagcg gtggcagcag cctaggttaa ttaagctgcg ctagtagacg
     3961 agtccatgtg ctggcgttca aatttcgcag cagcggtttc tttaccagac tcgagttatt
     4021 actttcgaga tttcttgcct cttcgtccct ttcgagattt cttgcctctt cgtcctttct
     4081 tgtacagctc gtccattccg tgagtaatgc ctgcagcggt cacgaattct aacagaacca
     4141 tgtgatcgcg tttttcgttc ggatctttgc tcagtgctga ctgggtgctc aggtagtggt
     4201 tgtcaggtaa cagtacaggc ccgtcaccaa tcggcgtgtt ttgttgataa tgatccgcta
     4261 attgaacgct gccgtcttcc acgttatggc gaattttgaa atttgccttg ataccgtttt
     4321 tctgtttatc ggccgtaata tacacgttat gagaattaaa gttatattcc agcttatggc
     4381 ctaaaatgtt gccgtcttct ttgaagtcga tacctttcag ttcgatacgg tttactaatg
     4441 tgtcgccttc aaatttcact tccgcacgcg ttttataggt gccatcgtcc ttgaagctga
     4501 ttgtacgctc ttgaacatat ccttcaggca tggctgattt gaaaaaatcg tgttgtttca
     4561 tatgatctgg gtaacgactg aagcactgaa cgccataggt cagggtggtc actaaggtgg
     4621 gccacggaac gggtaatttg cctgttgtgc aaataaactt cagggtcaat ttaccattgg
     4681 tagcgtcacc ttcgccttcg ccgcgcacgc taaatttgtg accgttcacg tcgccgtcta
     4741 attccactaa gatcggtact acacccgtaa acagctcttc acctttgcta gcgccaccgt
     4801 gatggtggtg gtgatgaccc atggtatatc tccttcttaa agttaaacaa aattatttct
     4861 agaggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa tttcgcggga
     4921 tcgagatcga tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca
     4981 ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac
     5041 ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga
     5101 ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc
     5161 aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagatccc
     5221 ggacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag
     5281 tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg
     5341 tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac
     5401 gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca
     5461 acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca
     5521 cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt
     5581 ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct
     5641 tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat
     5701 tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac
     5761 acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct
     5821 ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc
     5881 gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc
     5941 g
//
