LOCUS       sequence-405655-        5902 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 5902)
  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 5902)
  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)-H1
FEATURES             Location/Qualifiers
     source          1..5902
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     975..994
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1014..1091)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1297..1316
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1404..1423
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1404..1422
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1423..1447
                     /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             1462..1484
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1497..1514
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2271..2315
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2353..2371)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2367..2414
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2451..2906
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2538..2557)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2748..2769
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3021..3812,3813..3881))
                     /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     3644..3663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3882..3974)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4055..4643
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4544..4563
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4797..4814
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4829..4968
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4894..4913)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5054..5076
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5070..5261)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(5833..5902,1..1013))
                     /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"
ORIGIN
        1 gaggcggttt gcgtattggg cgccagggtg gtttttcttt tcaccagtga gacgggcaac
       61 agctgattgc ccttcaccgc ctggccctga gagagttgca gcaagcggtc cacgctggtt
      121 tgccccagca ggcgaaaatc ctgtttgatg gtggttaacg gcgggatata acatgagctg
      181 tcttcggtat cgtcgtatcc cactaccgag atgtccgcac caacgcgcag cccggactcg
      241 gtaatggcgc gcattgcgcc cagcgccatc tgatcgttgg caaccagcat cgcagtggga
      301 acgatgccct cattcagcat ttgcatggtt tgttgaaaac cggacatggc actccagtcg
      361 ccttcccgtt ccgctatcgg ctgaatttga ttgcgagtga gatatttatg ccagccagcc
      421 agacgcagac gcgccgagac agaacttaat gggcccgcta acagcgcgat ttgctggtga
      481 cccaatgcga ccagatgctc cacgcccagt cgcgtaccgt cttcatggga gaaaataata
      541 ctgttgatgg gtgtctggtc agagacatca agaaataacg ccggaacatt agtgcaggca
      601 gcttccacag caatggcatc ctggtcatcc agcggatagt taatgatcag cccactgacg
      661 cgttgcgcga gaagattgtg caccgccgct ttacaggctt cgacgccgct tcgttctacc
      721 atcgacacca ccacgctggc acccagttga tcggcgcgag atttaatcgc cgcgacaatt
      781 tgcgacggcg cgtgcagggc cagactggag gtggcaacgc caatcagcaa cgactgtttg
      841 cccgccagtt gttgtgccac gcggttggga atgtaattca gctccgccat cgccgcttcc
      901 actttttccc gcgttttcgc agaaacgtgg ctggcctggt tcaccacgcg ggaaacggtc
      961 tgataagaga caccggcata ctctgcgaca tcgtataacg ttactggttt cacattcacc
     1021 accctgaatt gactctcttc cgggcgctat catgccatac cgcgaaaggt tttgcgccat
     1081 tcgatggtgt ccgggatctc gacgctctcc cttatgcgac tcctgcatta ggaagcagcc
     1141 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcat gcaaggagat
     1201 ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct
     1261 catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc
     1321 agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga
     1381 gatcgatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     1441 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatatacc atgggtcatc
     1501 accaccacca tcacggtggc gctagcaaag gtgaacgtct gtttactggt gtagtaccga
     1561 tcttagtgga attagacggc gacgtgaacg gtcataaatt tagcgtgcgc ggcgagggcg
     1621 aaggtgacgc taccaatggt aaattgaccc tgaagtttat ttgcacaaca ggcaaattac
     1681 ccgttccgtg gcccacctta gtgaccaccc tgacctatgg cgttcagtgc ttcagtcgtt
     1741 accctaaaca tatgaaacaa cacgattttt tcaaatcagc catgcctgaa ggatatgttc
     1801 aagagcgtac aatcagcttc aagaaggatg gcacctataa aacgcgtgcg gaagtgaaat
     1861 ttgaaggccg cacattagta aatcgtatcg aactgaaagg tcgtgacttc aaagaagacg
     1921 gcaacatttt aggccataaa ctggaatata actttaattc tcataacgtg tatattacgg
     1981 ccgataaaca gaagaatggt atcaaggcaa atttcaaaat tcgccataac gtggaggacg
     2041 gcagcgttca attagcggat cattatcaac aaaacacgcc gattggtgat gggcctgtac
     2101 tgttacctcg caaccactac ctgagcaccc aatctgccct gagcaaagat ccgaaagaaa
     2161 aacgcgatca catggttctg ttagaattcg tgaccgctgc aggcattacg cacggaatgg
     2221 acgaacgcta caagaagcgt agacgcaaga aataataact cgagtctggt aaagaaaccg
     2281 ctgctgcgaa atttgaacgc cagcacatgg actcgtctac tagcgcagct taattaacct
     2341 aggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     2401 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
     2461 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     2521 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     2581 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     2641 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     2701 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     2761 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     2821 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     2881 taacaaaata ttaacgttta caatttctgg cggcacgatg gcatgagatt atcaaaaagg
     2941 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3001 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3061 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3121 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3181 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3241 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     3301 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     3361 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     3421 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     3481 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3541 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3601 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     3661 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3721 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3781 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3841 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatca
     3901 tgattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     3961 gaaaaataaa caaataggtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4021 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4081 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4141 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4201 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4261 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4321 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4381 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4441 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4501 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4561 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4621 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4681 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4741 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     4801 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     4861 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     4921 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     4981 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     5041 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5101 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     5161 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     5221 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     5281 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     5341 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     5401 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     5461 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     5521 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     5581 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     5641 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     5701 gacaggagca cgatcatgct agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     5761 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     5821 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5881 tgaatcggcc aacgcgcggg ga
//
