LOCUS       sequence-474421-        5963 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For overexpression of mCherry-RPL18.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5963)
  AUTHORS   Zhang Y, Stoppelkamp I, Fernandez-Pernas P, Allram M, Charman M, 
            Magalhaes AP, Piedavent-Salomon M, Sommer G, Sung YC, Meyer K, Grams
            N, Halko E, Dongre S, Meierhofer D, Malszycki M, Ilik IA, Aktas T, 
            Kraushar ML, Vastenhouw N, Weitzman MD, Grebien F, Niskanen H, Hnisz
            D
  TITLE     Probing condensate microenvironments with a micropeptide killswitch.
  JOURNAL   Nature. 2025 Jun 4. doi: 10.1038/s41586-025-09141-5.
  PUBMED    40468084
REFERENCE   2  (bases 1 to 5963)
  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: "Nature. 
            2025 Jun 4. doi: 10.1038/s41586-025-09141-5."
COMMENT     Sequence Label: pRK5_mCherry-RPL18
FEATURES             Location/Qualifiers
     source          1..5963
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      933..942
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2263..2272
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2276..2410
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2313..2332)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2367..2386
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2479..2836
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2479..2670
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2502..2522)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2687..2822
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2856..2873)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2856..2872)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2865..2887)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3085..3540
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3172..3191)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3382..3403
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3557..3576)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3676..3698
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3736..3754)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3822..3926
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3927..3995,3996..4787)
                     /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(4145..4164)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4958..5546
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5447..5466
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5700..5717
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5834..5855
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5870..5875,5876..5893,5894..5900)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5908..5924
                     /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     5913..5935
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5932..5948
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5932..5948
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc tatcgattga attcgtaccg gtgccaccat ggtttccaaa ggagaggagg
      961 ataatatggc tataattaaa gagtttatgc ggttcaaggt gcatatggag ggatctgtga
     1021 acggtcacga gttcgaaatc gagggcgaag gcgaggggcg accctatgaa gggacacaga
     1081 cggccaaact gaaggtgacc aagggtggcc ccctgccttt cgcctgggac atcctgtcac
     1141 cccagtttat gtacggctct aaggcttacg ttaagcaccc tgccgatatt ccggactact
     1201 tgaaactgag cttccctgaa gggttcaagt gggaacgggt aatgaatttt gaggatggcg
     1261 gtgtagtcac agttacccag gacagctctc ttcaggacgg agaatttatc tataaggtta
     1321 aacttcgggg cactaacttc ccatcagacg gccccgtgat gcagaagaaa actatgggct
     1381 gggaagccag ctcagaacgc atgtatcccg aggacggagc actgaaggga gaaatcaagc
     1441 agcgactcaa gctgaaggat gggggacatt atgatgctga ggtcaaaacc acctataagg
     1501 ccaagaaacc cgttcagctt cctggggcat acaacgtgaa tatcaaactg gatattacat
     1561 cccataacga agactatacc atcgtggaac agtacgagcg ggccgagggc agacatagca
     1621 cagggggcat ggatgaattg tacaaggggg gaggcggcag tggcggaggg ggatctatgg
     1681 gagtggacat ccgccataac aaggaccgaa aggttcggcg caaggagccc aagagccagg
     1741 atatctacct gaggctgttg gtcaagttat acaggtttct ggccagaaga accaactcca
     1801 cattcaacca ggttgtgttg aagaggttgt ttatgagtcg caccaaccgg ccgcctctgt
     1861 ccctttcccg gatgatccgg aagatgaagc ttcctggccg ggaaaacaag acggccgtgg
     1921 ttgtggggac cataactgat gatgtgcggg ttcaggaggt acccaaactg aaggtatgtg
     1981 cactgcgcgt gaccagccgg gcccgcagcc gcatcctcag ggcagggggc aagatcctca
     2041 ctttcgacca gctggccctg gactccccta agggctgtgg cactgtcctg ctctccggtc
     2101 ctcgcaaggg ccgagaggtg taccggcatt tcggcaaggc cccaggaacc ccgcacagcc
     2161 acaccaaacc ctacgtccgc tccaagggcc ggaagttcga gcgtgccaga ggccgacggg
     2221 ccagccgagg ctacaaaaac tgagtcgacc tgcagaagct tggccgccat ggcccaactt
     2281 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     2341 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     2401 tgtctggatc gggaattaat tcggcgcagc accatggcct gaaataacct ctgaaagagg
     2461 aacttggtta ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     2521 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2581 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2641 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2701 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2761 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2821 cctaggcttt tgcaaaaagc tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt
     2881 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     2941 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     3001 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3061 cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3121 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3181 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3241 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3301 tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3361 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3421 tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3481 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt
     3541 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc
     3601 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     3661 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     3721 gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     3781 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3841 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3901 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3961 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     4021 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     4081 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     4141 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     4201 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     4261 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     4321 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     4381 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4441 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4501 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4561 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4621 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4681 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4741 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     4801 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     4861 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     4921 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     4981 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     5041 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     5101 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     5161 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     5221 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     5281 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     5341 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     5401 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     5461 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     5521 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     5581 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     5641 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     5701 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     5761 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     5821 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     5881 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     5941 gctatgacat gattacgaat taa
//
