LOCUS       sequence-374359-        6711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression mCherry-Elk1DM in yeast cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6711)
  AUTHORS   Sang D, Shu T, Pantoja CF, Ibanez de Opakua A, Zweckstetter M, Holt 
            LJ
  TITLE     Condensed-phase signaling can expand kinase specificity and respond 
            to macromolecular crowding.
  JOURNAL   Mol Cell. 2022 Oct 6;82(19):3693-3711.e10. doi: 
            10.1016/j.molcel.2022.08.016. Epub 2022 Sep 14.
  PUBMED    36108633
REFERENCE   2  (bases 1 to 6711)
  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"; doi: 
            "10.1016/j.molcel.2022.08.016"; journalName: "Mol Cell"; date: 
            "2022-10-6- 6"; volume: "82"; issue: "19"; pages: "3693-3711.e10"
COMMENT     Sequence Label: pAV103-PRNR2-mCherry-Elk1DM-1SIM
FEATURES             Location/Qualifiers
     source          1..6711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(26..454)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(136..157)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     348..367
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     580..602
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     594..611
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     595..611
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     621..640
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        621..639
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1411..1437
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             1495..2202
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1640..1658)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1908..1927
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2102..2125
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3083..3103)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3083..3101)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3122..3138)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3122..3138)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3135..3157)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3146..3162
                     /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)."
     promoter        complement(join(3170..3176,3177..3194,3195..3200))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3215..3236
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3353..3370)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3524..4112)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3604..3623)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4283..5074,5075..5143))
                     /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     4906..4925
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5144..5248)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5316..5334
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     5434..5453
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5600..5787
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             5788..6450
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
ORIGIN
        1 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
       61 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
      121 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
      181 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
      241 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
      301 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
      361 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
      421 cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg ctgcgcaact
      481 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
      541 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
      601 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc
      661 ccctcgagag tcgaacaaga agcaggcaaa gtttagagca ctgcccctcc gcactcaaaa
      721 aagaaaaaac taggaggaaa ataaaattct caaccacaca aacacataaa cacatacaaa
      781 tacaaataca agcttattta cttgacatcg cgcgatcttc cactattcag cgccgtccgc
      841 cctctctcgt gttttttgtt tacgcgacaa ctatgcgaaa tccggagcaa cgggcaaccg
      901 tttggggaaa gaccacaccc acgcgcgatc gccatggcaa cgaggtcgca cacgccccac
      961 acccagacct ccctgcgagc gggcatgggt acaatgtccc cgttgccaca gagaccactt
     1021 cgtagcacag cgcagagcgt agcgtgttgt tgctgctgac aaaagaaaat ttttcttagc
     1081 aaagcaaagg aggggaagca cgggcagata gcaccgtacc atacccttgg aaactcgaaa
     1141 tgaacgaagc aggaaatgag agaatgagag ttttgtaggt atatatagcg gtagtgtttg
     1201 cgcgttacca tcatcttctg gatctatcta ttgttctttt cctcatcact ttcccctttt
     1261 tcgctcttct tcttgtcttt tatttctttc ttttttttaa ttgttccctc gattggctat
     1321 ctaccaaaga atccaaactt aatacacgta tttatttgtc caattaccga attcatgttg
     1381 atcacaagtg gagcgtcagg ggcacttcag ttgccaccgt tggaaagact aactctgggc
     1441 tccgctggat cagctttagc cttgaaatta gcaggtcttg atatcctgat cactatggtg
     1501 agcaagggcg aggaggacaa catggccatc atcaaggagt tcatgcgctt caaggtgcac
     1561 atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc
     1621 tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc
     1681 tgggacatcc tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc
     1741 gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
     1801 aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag
     1861 ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag
     1921 aagaagacca tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg
     1981 aagggcgaga tcaagcagag gctgaagctg aaggacggcg gccactacga cgccgaggtc
     2041 aagaccacct acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc
     2101 aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc
     2161 gagggccgcc actccaccgg cggcatggac gagctgtaca aggcggccgc aggtggttcc
     2221 gggggttcag ggggttctgg tggaagcgcc tgcctggaag cggaagaagc agggctgccc
     2281 ttgcaagtta ttcttactcc cccggaggct cccaatctta aatctgaaga gcttaacgtc
     2341 gagccgggac tgggacgtgc gcttcctcca gaagttaaag tggagggtcc caaagaagaa
     2401 ttagaggtcg ctggagaacg cggctttgtt cccgagacta cgaaagccga gcctgaggta
     2461 cctccgcaag aaggagtccc agctcgctta ccggccgtag tgatggatac tgctggacag
     2521 gcaggcggtc atgctgcatc tagtccagaa atcagtcaac cgcaagcggg ggccgcgccc
     2581 cgtgatgcag cggcgcctct gtcgccgtcg cttcttggag gacctggccc agagcgcacc
     2641 cccggcagcg ggtctggttc tggattgcag gcgccggggc cagccctgac tccttcctta
     2701 ttgcctactc acacgttgac acccgtctta ttgaccccgt ccagcttgcc tccatcaatc
     2761 cacttttggt ctacccttag cccgattgcc ccacgctctc ccgcaaaatt gagtgcagcg
     2821 gcacccagca gcggatccgc tcaagttcac atcccctcca tttctgtgga cggattaagt
     2881 acgccagtcg tgcttagccc aggtcctcaa aaaccaacta gaggcggctc gggaggttcg
     2941 ggcggcagcg gcggaagcaa agtcgatgta attgacttaa cgatcgaatc tagcagcgat
     3001 gaagaagaag atccaccggc taaacgttag ttaattaata tataactgaa taaatttttt
     3061 caaagagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
     3121 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     3181 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     3241 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     3301 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     3361 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3421 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3481 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3541 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3601 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3661 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3721 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3781 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3841 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     3901 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3961 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4021 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4081 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4141 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4201 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4261 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4321 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     4381 gggagggctt accatctggc cccagtgctg caatgatacc gcgagatcca cgctcaccgg
     4441 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     4501 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     4561 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     4621 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     4681 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     4741 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     4801 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     4861 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     4921 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     4981 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5041 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5101 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5161 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5221 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     5281 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttg
     5341 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     5401 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     5461 tgcaccataa attcccgttt taagagcttg gtgagcgcta ggagtcactg ccaggtatcg
     5521 tttgaacacg gcattagtca gggaagtcat aacacagtcc tttcccgcaa ttttcttttt
     5581 ctattactct tggcctcctc tagtacactc tatatttttt tatgcctcgg taatgatttt
     5641 catttttttt tttccaccta gcggatgact cttttttttt cttagcgatt ggcattatca
     5701 cataatgaat tatacattat ataaagtaat gtgatttctt cgaagaatat actaaaaaat
     5761 gagcaggcaa gataaacgaa ggcaaagatg acagagcaga aagccctagt aaagcgtatt
     5821 acaaatgaaa ccaagattca gattgcgatc tctttaaagg gtggtcccct agcgatagag
     5881 cactcgatct tcccagaaaa agaggcagaa gcagtagcag aacaggccac acaatcgcaa
     5941 gtgattaacg tccacacagg tatagggttt ctggaccata tgatacatgc tctggccaag
     6001 cattccggct ggtcgctaat cgttgagtgc attggtgact tacacataga cgaccatcac
     6061 accactgaag actgcgggat tgctctcggt caagctttta aagaggccct aggggccgtg
     6121 cgtggagtaa aaaggtttgg atcaggattt gcgcctttgg atgaggcact ttccagagcg
     6181 gtggtagatc tttcgaacag gccgtacgca gttgtcgaac ttggtttgca aagggagaaa
     6241 gtaggagatc tctcttgcga gatgatcccg cattttcttg aaagctttgc agaggctagc
     6301 agaattaccc tccacgttga ttgtctgcga ggcaagaatg atcatcaccg tagtgagagt
     6361 gcgttcaagg ctcttgcggt tgccataaga gaagccacct cgcccaatgg taccaacgat
     6421 gttccctcca ccaaaggtgt tcttatgtag tgacaccgat tatttaaagc tgcagcatac
     6481 gatatatata catgtgtata tatgtatacc tatgaatgtc agtaagtatg tatacgaaca
     6541 gtatgatact gaagatgaca aggtaatgca tcattctata cgtgtcattc tgaacgaggc
     6601 gcgctttcct tttttctttt tgctttttct ttttttttct cttgaactcg acggatctat
     6661 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga a
//
