LOCUS       sequence-351690-        5691 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Mammalian expression of mCherry-FKBP-TEVprotease(Delta-220-242).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5691)
  AUTHORS   Yoshikawa M, Yoshii T, Ikuta M, Tsukiji S
  TITLE     Synthetic Protein Condensates That Inducibly Recruit and Release 
            Protein Activity in Living Cells.
  JOURNAL   J Am Chem Soc. 2021 May 5;143(17):6434-6446. doi: 
            10.1021/jacs.0c12375. Epub 2021 Apr 23.
  PUBMED    33890764
REFERENCE   2  (bases 1 to 5691)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/jacs.0c12375"; journalName: "J Am Chem Soc"; date: 
            "2021-05-5- 5"; volume: "143"; issue: "17"; pages: "6434-6446"
COMMENT     Sequence Label: pCMV-mCherry-FKBP-TEVp
FEATURES             Location/Qualifiers
     source          1..5691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..354
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        355..558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     509..529
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             603..1310
                     /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(748..766)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1016..1035
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(1332..1334,1335..1655)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="MGVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNK
                     PFKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVEL
                     LKLE"
     polyA_signal    2469..2590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2506..2525)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2560..2579
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2597..3052)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2734..2755)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2946..2965
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3079..3183
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3185..3542
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3185..3376
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3208..3228)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3393..3528
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3577..4371
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3577..4371
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3631..3650)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4241..4260
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4559..4578)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4603..4650
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4979..5567
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5468..5487
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
       61 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      121 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      181 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      241 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      301 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      361 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      421 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      481 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      541 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggtcgcca
      601 ccatggtgag caagggcgag gaggataaca tggccatcat caaggagttc atgcgcttca
      661 aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
      721 gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt ggccccctgc
      781 ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc tacgtgaagc
      841 accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc aagtgggagc
      901 gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg
      961 acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg
     1021 taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     1081 gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc cactacgacg
     1141 ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacaacg
     1201 tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     1261 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag tccggactca
     1321 gatctcgagc tatgggagtg caggtggaaa ccatctcccc aggagacggg cgcaccttcc
     1381 ccaagcgcgg ccagacctgc gtggtgcact acaccgggat gcttgaagat ggaaagaaat
     1441 ttgattcctc ccgggacaga aacaagccct ttaagtttat gctaggcaag caggaggtga
     1501 tccgaggctg ggaagaaggg gttgcccaga tgagtgtggg tcagagagcc aaactgacta
     1561 tatctccaga ttatgcctat ggtgccactg ggcacccagg catcatccca ccacatgcca
     1621 ctctcgtctt cgatgtggag cttctaaaac tggaagaatt ctgcagtcga cggtaccgca
     1681 tgggcgagag ccttttcaag ggcccgaggg actacaaccc gatctccagc accatctgtc
     1741 acctgaccaa cgagagcgac ggtcacacca ctagtctgta cggcatcggc ttcggcccct
     1801 tcatcatcac caacaagcat ctgttcagga ggaataacgg cacactgctg gtgcaaagcc
     1861 tgcacggcgt gttcaaagtg aagaacacaa ccaccctgca acagcacctg atcgacggca
     1921 gggacatgat tatcatcagg atgcccaagg acttcccccc ctttccccag aaactgaagt
     1981 tcagggagcc acaaagggag gagcgaatct gcctggtgac caccaacttc cagaccaagt
     2041 ccatgagcag catggtctct gataccagct gcaccttccc cagcagcgac ggcatcttct
     2101 ggaagcactg gattcagacg aaggatggcc aatgcggcag cccattggtg agcactaggg
     2161 acggcttcat cgtgggcatc cacagcgcca gcaattttac caataccaac aactacttca
     2221 cgagcgtgcc gaaaaacttc atggagctgt tgaccaatca agaggcgcag cagtgggtga
     2281 gcggctggag gctgaacgcc gacagcgttc tttggggcgg acataaggtg ttcatggtcg
     2341 gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta gaggttttac
     2401 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2461 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2521 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2581 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2641 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     2701 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     2761 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     2821 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     2881 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     2941 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3001 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3061 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3121 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3181 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3241 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3301 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3361 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3421 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3481 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3541 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3601 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3661 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     3721 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     3781 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     3841 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     3901 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     3961 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4021 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4081 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4141 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4201 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4261 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4321 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4381 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4441 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4501 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     4561 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4621 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4681 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     4741 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     4801 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     4861 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     4921 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     4981 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5041 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5101 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5161 agaactctgt atcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5221 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5281 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5341 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5401 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5461 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5521 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5581 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5641 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta a
//
