LOCUS       sequence-382959-        5908 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  antibiotic resistance enzyme with cationic peptide tag to promote 
            biomolecular condensate formation in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5908)
  AUTHORS   Yeong V, Wang JW, Horn JM, Obermeyer AC
  TITLE     Intracellular phase separation of globular proteins facilitated by 
            short cationic peptides.
  JOURNAL   Nat Commun. 2022 Dec 22;13(1):7882. doi: 10.1038/s41467-022-35529-2.
  PUBMED    36550144
REFERENCE   2  (bases 1 to 5908)
  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: "Nat 
            Commun."; date: "2022-12-22"; volume: "13(1):7882. doi"; pages: " 
            10.1038/s41467-022-35529-2"
COMMENT     Sequence Label: pVY-12 CAT-tag3
FEATURES             Location/Qualifiers
     source          1..5908
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     158..180
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(174..365)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(937..2019)
                     /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     1981..2000
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2020..2097)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2303..2322
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2410..2429
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2410..2428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2429..2453
                     /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             2468..2490
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2503..2520
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2530..3183
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="EKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFLK
                     TVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSSL
                     WSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANMD
                     NFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2593..2612)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3283..3327
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(3365..3383)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3379..3426
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3463..3918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3550..3569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3760..3781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(4033..4824,4825..4893))
                     /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     4656..4675
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4894..4986)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5067..5655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5556..5575
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5809..5826
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(5841..5908,1..72)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(join(5906..5908,1..17))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
       61 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
      121 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
      181 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
      241 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
      301 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
      361 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
      421 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt
      481 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
      541 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
      601 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
      661 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
      721 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
      781 cgccagccta gccgggtcct caacgacagg agcacgatca tgctagtcat gccccgcgcc
      841 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
      901 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
      961 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     1021 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     1081 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     1141 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     1201 gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     1261 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     1321 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     1381 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     1441 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     1501 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     1561 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     1621 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     1681 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     1741 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     1801 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     1861 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     1921 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     1981 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     2041 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     2101 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     2161 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     2221 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     2281 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     2341 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc
     2401 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
     2461 aaataatttt gtttaacttt aagaaggaga tataccatgg gtcatcacca ccaccatcac
     2521 ggtggcgctg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     2581 aaagaacatt ttgaggcatt tcagtcagtt gctcaatgta cctataacca gaccgttcag
     2641 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     2701 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa
     2761 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     2821 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     2881 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     2941 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     3001 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     3061 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ctgtgatggc
     3121 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg
     3181 gcgggaggaa gcaagaaacg caagaagcgt ggcggtagca aaaagcgtaa gaagcgtgga
     3241 ggttcaaaga aacgtaaaaa gcgataataa ctcgagtctg gtaaagaaac cgctgctgcg
     3301 aaatttgaac gccagcacat ggactcgtct actagcgcag cttaattaac ctaggctgct
     3361 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     3421 tttttgctga aaggaggaac tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg
     3481 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3541 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3601 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3661 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3721 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3781 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3841 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     3901 tattaacgtt tacaatttct ggcggcacga tggcatgaga ttatcaaaaa ggatcttcac
     3961 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4021 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4081 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4141 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4201 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4261 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4321 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4381 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4441 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     4501 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     4561 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     4621 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     4681 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4741 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4801 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4861 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat catgattgaa
     4921 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     4981 aacaaatagg tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5041 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5101 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5161 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     5221 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5281 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5341 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5401 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5461 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5521 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5581 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5641 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5701 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5761 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5821 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     5881 tcacaccgca atggtgcact ctcagtac
//
