LOCUS       sequence-405643-        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(0)-L1
FEATURES             Location/Qualifiers
     source          1..5911
                     /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             877..921
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(959..977)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      973..1020
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1057..1512
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1144..1163)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1354..1375
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(1627..2418,2419..2487))
                     /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     2250..2269
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2488..2580)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2661..3249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3150..3169
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3403..3420
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3435..3574
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3500..3519)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3660..3682
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3676..3867)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4439..5521)
                     /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     5483..5502
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5522..5599)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5805..5824
                     /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 ctgatcatat gaaacaacac
      361 gattttttca aatcagccat gcctgaagga tatgttcaag agcgtacaat cagcttcaag
      421 gacgatggca cctataaaac gcgtgcggaa gtgaaatttg aaggcgacac 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 cctgtactgt tacctcgcaa ccactacctg
      721 agcacccaat ctgccctgag caaagatccg aaagaaaaac gcgatcacat ggttctgtta
      781 gaattcgtga ccactgcagg cattacgcac ggaatggacg aacgctacaa gggtcgaaga
      841 gggaagaaat cgcggaagta ataactcgag tctggtaaag aaaccgctgc tgcgaaattt
      901 gaacgccagc acatggactc gtctactagc gcagcttaat taacctaggc tgctgccacc
      961 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     1021 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa
     1081 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     1141 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     1201 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     1261 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     1321 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     1381 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     1441 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     1501 cgtttacaat ttctggcggc acgatggcat gagattatca aaaaggatct tcacctagat
     1561 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1621 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1681 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1741 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1801 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1861 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1921 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1981 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     2041 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     2101 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     2161 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     2221 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2281 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     2341 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2401 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2461 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatcatgat tgaagcattt
     2521 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     2581 taggtcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     2641 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     2701 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     2761 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     2821 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     2881 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     2941 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     3001 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     3061 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     3121 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     3181 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     3241 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     3301 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     3361 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     3421 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3481 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     3541 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     3601 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3661 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     3721 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     3781 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     3841 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     3901 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     3961 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     4021 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     4081 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     4141 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     4201 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     4261 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     4321 catgctagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     4381 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     4441 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4501 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     4561 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     4621 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     4681 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc
     4741 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     4801 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     4861 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     4921 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     4981 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     5041 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     5101 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     5161 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     5221 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     5281 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     5341 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     5401 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     5461 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     5521 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     5581 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     5641 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     5701 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     5761 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     5821 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     5881 aggatcgaga tcgatctcga tcccgcgaaa t
//
