LOCUS       sequence-405640-        5905 bp    DNA     circular SYN 15-APR-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 5905)
  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 5905)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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..5905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    104..243
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     159..178
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(258..275)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(429..1017)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(509..528)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1098..1190
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1191..1259,1260..2051)
                     /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(1409..1428)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2166..2621)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2303..2324)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2515..2534
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(2658..2705)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2701..2719
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2757..2801)
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             complement(3561..3578)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3591..3613
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3628..3652
                     /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(3652..3671)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3653..3671)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3759..3778)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3984..4061
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4062..5144
                     /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(4081..4100)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             join(5716..5905,1..2)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(join(5901..5905,1..18))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
       61 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
      121 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
      181 gagagtgcac cattgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
      241 aggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
      301 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
      361 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
      421 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
      481 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
      541 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
      601 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
      661 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
      721 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      781 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      841 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
      901 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      961 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1021 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     1081 attttggtca tgacctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1141 gagacaataa ccctgataaa tgcttcaatc atgattgaaa aaggaagagt atgagtattc
     1201 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1261 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1321 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1381 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     1441 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1501 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1561 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     1621 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     1681 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     1741 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     1801 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     1861 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     1921 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     1981 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2041 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     2101 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg ccatcgtgcc
     2161 gccagaaatt gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2221 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2281 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2341 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     2401 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2461 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     2521 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     2581 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc caatccggat
     2641 atagttcctc ctttcagcaa aaaacccctc aagacccgtt tagaggcccc aaggggttat
     2701 gctagttatt gctcagcggt ggcagcagcc taggttaatt aagctgcgct agtagacgag
     2761 tccatgtgct ggcgttcaaa tttcgcagca gcggtttctt taccagactc gagttattat
     2821 ttcttccttc tcctcttttt cttgtacagc tcgtccattc cgtgagtaat gcctgcagcg
     2881 gtcacgaatt ctaacagaac catgtgatcg cgtttttcgt tcggatcttt gctcagtgct
     2941 gactgggtgc tcaggtagtg gttgtcaggt aacagtacag gcccgtcacc aatcggcgtg
     3001 ttttgttgat aatgatccgc taattgaacg ctgccgtctt ccacgttatg gcgaattttg
     3061 aaatttgcct tgataccgtt tttctgttta tcggccgtaa tatacacgtt atgagaatta
     3121 aagttatatt ccagcttatg gcctaaaatg ttgccgtctt ctttgaagtc gatacctttc
     3181 agttcgatac ggtttactaa tgtgtcgcct tcaaatttca cttccgcacg cgttttatag
     3241 gtgccatcgt ccttgaagct gattgtacgc tcttgaacat atccttcagg catggctgat
     3301 ttgaaaaaat cgtgttgttt catatgatct gggtaacgac tgaagcactg aacgccatag
     3361 gtcagggtgg tcactaaggt gggccacgga acgggtaatt tgcctgttgt gcaaataaac
     3421 ttcagggtca atttaccatt ggtagcgtca ccttcgcctt cgccgcgcac gctaaatttg
     3481 tgaccgttca cgtcgccgtc taattccact aagatcggta ctacacccgt aaacagctct
     3541 tcacctttgc tagcgccacc gtgatggtgg tggtgatgac ccatggtata tctccttctt
     3601 aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt cccctatagt
     3661 gagtcgtatt aatttcgcgg gatcgagatc gatctcgatc ctctacgccg gacgcatcgt
     3721 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     3781 tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt
     3841 ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac cattccttgc
     3901 ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc aggagtcgca
     3961 taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt cgcggtatgg
     4021 catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat
     4081 acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg
     4141 ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt
     4201 acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg
     4261 ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg
     4321 ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct
     4381 gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc
     4441 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     4501 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     4561 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     4621 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     4681 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     4741 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     4801 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     4861 acatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     4921 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     4981 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     5041 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     5101 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     5161 gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac
     5221 ccagtcagct ccttccggtg ggcgcggggc atgactagca tgatcgtgct cctgtcgttg
     5281 aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc accgatacgc
     5341 gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg
     5401 gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt
     5461 atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta
     5521 ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg catccatacc
     5581 gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc agtaacccgt
     5641 atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag aaatccccct
     5701 tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc ccgctttatc
     5761 agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga tgaacaggca
     5821 gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg cctcgcgcgt
     5881 ttcggtgatg acggtgaaaa cctct
//
