LOCUS       sequence-405654-        5965 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 5965)
  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 5965)
  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..5965
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             94..111
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             931..975
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1013..1031)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1027..1074
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1111..1566
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1198..1217)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1408..1429
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1681..2472,2473..2541))
                     /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     2304..2323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2542..2634)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2715..3303
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3204..3223
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3457..3474
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3489..3628
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3554..3573)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3714..3736
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3730..3921)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4493..5575)
                     /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     5537..5556
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5576..5653)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5859..5878
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggtcatcacc accaccatca cggtggcgct
      121 agcaaaggtg aacgtctgtt tactggtgta gtaccgatct tagtggaatt agacggcgac
      181 gtgaacggtc ataaatttag cgtgcgcggc gagggcgaag gtgacgctac caatggtaaa
      241 ttgaccctga agtttatttg cacaacaggc aaattacccg ttccgtggcc caccttagtg
      301 accaccctga cctatggcgt tcagtgcttc agtcgttacc ctaaacatat gaaacaacac
      361 gattttttca aatcagccat gcctgaagga tatgttcaag agcgtacaat cagcttcaag
      421 aaggatggca cctataaaac gcgtgcggaa gtgaaatttg aaggccgcac attagtaaat
      481 cgtatcgaac tgaaaggtcg tgacttcaaa gaagacggca acattttagg ccataaactg
      541 gaatataact ttaattctca taacgtgtat attacggccg ataaacagaa gaatggtatc
      601 aaggcaaatt tcaaaattcg ccataacgtg gaggacggca gcgttcaatt agcggatcat
      661 tatcaacaaa acacgccgat tggtgatggg cctatactgt tacctcgcaa ccactacctg
      721 agcacccaat ctgccctgag caaagatccg aaagaaaaac gcgatcacat ggttctgtta
      781 gaattcgtga ccgctgcagg cattacgcac ggaatggacg aacgctacaa gggtcgaaga
      841 gggaagaaat cgcggaaggg tcgaagaggg aagaaatcgc ggaagggaag aaggggtaag
      901 aaatctagga agtaataact cgagtctggt aaagaaaccg ctgctgcgaa atttgaacgc
      961 cagcacatgg actcgtctac tagcgcagct taattaacct aggctgctgc caccgctgag
     1021 caataactag cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa
     1081 ggaggaacta tatccggatt ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     1141 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     1201 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     1261 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     1321 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     1381 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     1441 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     1501 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     1561 caatttctgg cggcacgatg gcatgagatt atcaaaaagg atcttcacct agatcctttt
     1621 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1681 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1741 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1801 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     1861 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1921 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1981 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2041 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2101 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2161 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     2221 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     2281 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     2341 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     2401 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     2461 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     2521 acggaaatgt tgaatactca tactcttcct ttttcaatca tgattgaagc atttatcagg
     2581 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaataggtc
     2641 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     2701 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     2761 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     2821 aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag
     2881 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     2941 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     3001 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     3061 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     3121 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     3181 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     3241 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     3301 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     3361 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     3421 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     3481 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     3541 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     3601 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     3661 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     3721 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     3781 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     3841 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     3901 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     3961 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     4021 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     4081 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     4141 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     4201 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     4261 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     4321 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgct
     4381 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     4441 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     4501 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4561 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     4621 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     4681 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     4741 tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc agcccggact
     4801 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     4861 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     4921 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     4981 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     5041 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     5101 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     5161 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     5221 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     5281 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     5341 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     5401 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     5461 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     5521 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     5581 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     5641 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     5701 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     5761 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     5821 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     5881 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     5941 gagatcgatc tcgatcccgc gaaat
//
