LOCUS       sequence-405653-        5938 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 5938)
  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 5938)
  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..5938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1437..1628
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1622..1644)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1730..1869
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1785..1804
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1884..1901)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2055..2643)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2135..2154)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2724..2816
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2817..2885,2886..3677)
                     /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(3035..3054)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3792..4247)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3929..3950)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4141..4160
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4284..4331)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4327..4345
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4383..4427)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(5220..5237)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             5250..5272
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    5287..5311
                     /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(5311..5330)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5312..5330)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5418..5437)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5643..5720
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5721..5938,1..865)
                     /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(5740..5759)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag
       61 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa
      121 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc
      181 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca
      241 gacacccatc aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca
      301 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc
      361 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat
      421 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct
      481 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc
      541 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gacatctcgg tagtgggata
      601 cgacgatacc gaagacagct catgttatat cccgccgtta accaccatca aacaggattt
      661 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt
      721 gaagggcaat cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa
      781 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
      841 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt
      901 aggcaccggg atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt
      961 gggcgcgggg catgactagc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc
     1021 ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact
     1081 gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt
     1141 cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg
     1201 caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt
     1261 gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac
     1321 aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc
     1381 gtttcatcgg tatcattacc cccatgaaca gaaatccccc ttacacggag gcatcagtga
     1441 ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc
     1501 ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc
     1561 acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa
     1621 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     1681 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     1741 cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat
     1801 tgtactgaga gtgcaccatt gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     1861 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     1921 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     1981 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2041 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2101 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2161 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2221 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2281 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2341 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2401 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2461 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     2521 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2581 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2641 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     2701 taagggattt tggtcatgac ctatttgttt atttttctaa atacattcaa atatgtatcc
     2761 gctcatgaga caataaccct gataaatgct tcaatcatga ttgaaaaagg aagagtatga
     2821 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2881 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2941 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3001 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3061 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3121 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3181 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3241 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3301 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3361 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3421 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3481 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3541 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3601 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3661 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3721 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgccat
     3781 cgtgccgcca gaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3841 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3901 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3961 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     4021 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     4081 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     4141 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     4201 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccaat
     4261 ccggatatag ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg
     4321 ggttatgcta gttattgctc agcggtggca gcagcctagg ttaattaagc tgcgctagta
     4381 gacgagtcca tgtgctggcg ttcaaatttc gcagcagcgg tttctttacc agactcgagt
     4441 tattacttcc gcgatttctt ccctcttcga cccttccgcg atttcttccc tcttcgaccc
     4501 ttgtagcgtt cgtccattcc gtgcgtaatg cctgcagcgg tcacgaattc taacagaacc
     4561 atgtgatcgc gtttttcttt cggatctttg ctcagggcag attgggtgct caggtagtgg
     4621 ttgcgaggta acagtacagg cccatcacca atcggcgtgt tttgttgata atgatccgct
     4681 aattgaacgc tgccgtcctc cacgttatgg cgaattttga aatttgcctt gataccattc
     4741 ttctgtttat cggccgtaat atacacgtta tgagaattaa agttatattc cagtttatgg
     4801 cctaaaatgt tgccgtcttc tttgaagtca cgacctttca gttcgatacg atttactaat
     4861 gtgcggcctt caaatttcac ttccgcacgc gttttatagg tgccatcctt cttgaagctg
     4921 attgtacgct cttgaacata tccttcaggc atggctgatt tgaaaaaatc gtgttgtttc
     4981 atatgtttag ggtaacgact gaagcactga acgccatagg tcagggtggt cactaaggtg
     5041 ggccacggaa cgggtaattt gcctgttgtg caaataaact tcagggtcaa tttaccattg
     5101 gtagcgtcac cttcgccctc gccgcgcacg ctaaatttat gaccgttcac gtcgccgtct
     5161 aattccacta agatcggtac tacaccagta aacagacgtt cacctttgct agcgccaccg
     5221 tgatggtggt ggtgatgacc catggtatat ctccttctta aagttaaaca aaattatttc
     5281 tagaggggaa ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg
     5341 atcgagatcg atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
     5401 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
     5461 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
     5521 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct
     5581 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc
     5641 cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     5701 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     5761 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     5821 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     5881 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccct
//
