LOCUS       sequence-485431-        8680 bp    DNA     circular SYN 05-SEP-2025
DEFINITION  Proximity-triggered Protein Trans-Splicing between mVenus and RFP, 
            Figure 1 in Nature Chemical Biology 20.10 (2024): 1353-1360..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8680)
  AUTHORS   Lee G, Muir TW
  TITLE     Distinct phases of cellular signaling revealed by time-resolved 
            protein synthesis.
  JOURNAL   bioRxiv [Preprint]. 2023 Jul 10:2023.07.10.548208. doi: 
            10.1101/2023.07.10.548208.
  PUBMED    37503273
REFERENCE   2  (bases 1 to 8680)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2023 Jul 10:2023.07.10.548208. doi: 
            10.1101/2023.07.10.548208."
COMMENT     Sequence Label: pLentiLox7.0-mVenus-CAGEn-Caax (Nrdj1)
FEATURES             Location/Qualifiers
     source          1..8680
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2616..2928
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     2847..2867
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     2945..2961
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3003..3036)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     enhancer        3152..3455
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3456..3659
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      3730..3739
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3766..3789
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3799..3822
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3832..3855
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(3871..3873,3874..3876,3877..4587)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFDGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3916..3937)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4177..4196)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4524..4545
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             5071..5391
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     protein_bind    complement(5482..5515)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5571..6157
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5624..5644)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6042..6053)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6160..6176)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6686..6866
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6928..7516)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7008..7027)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7687..8478,8479..8547))
                     /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     8310..8329
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8548..8652)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgcgct ctagagatcc gacgcgccat ctctaggccc
     2641 gcgccggccc cctcgcacag acttgtggga gaagctcggc tactcccctg ccccggttaa
     2701 tttgcatata atatttccta gtaactatag aggcttaatg tgcgataaaa gacagataat
     2761 ctgttctttt taatactagc tacattttac atgataggct tggatttcta taagagatac
     2821 aaatactaaa ttattatttt aaaaaacagc acaaaaggaa actcacccta actgtaaagt
     2881 aattgtgtgt tttgagacta taaatatccc ttggagaaaa gccttgttaa cgcgcggtga
     2941 ccctcgaggt cgacggtatc gataagctcg cttcacgaga ttccagcagg tcgagggacc
     3001 taataacttc gtatagcata cattatacga agttatatta agggttccaa gcttaagcgg
     3061 ccgcgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
     3121 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     3181 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     3241 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     3301 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     3361 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     3421 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     3481 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     3541 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     3601 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
     3661 gtttagtgaa ccgtcagatc cgctagaatt ctgcagtcga cggtaccgcg ggcccgggat
     3721 ccaccggtcg ccaccatggg tagtctcgag ggtaccatgg gttcagacta taaagatgat
     3781 gatgataaag gtggctccga ctacaaggat gacgatgaca agggaggtag tgactacaaa
     3841 gacgatgacg acaagggcgg aggaggaagt atggtgagca agggcgagga gctgttcacc
     3901 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3961 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt gatctgcacc
     4021 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctcggcta cggcctgcag
     4081 tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     4141 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     4201 gccgaggtga agttcgatgg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     4261 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     4321 gtctatatca ccgccgacaa gcagaagaac ggcatcaagg caaacttcaa gatccgccac
     4381 aacatcgagg acggcggcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     4441 gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa actgagcaaa
     4501 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     4561 actctcggca tggacgagct gtacaagaat ccatgctgtc tggtcggctc tagcgaaatc
     4621 atcacacgta attatgggaa gacaaccatt aaggaagtag ttgagatatt tgacaatgat
     4681 aaaaatatcc aagttctagc gtttaacact cacactgaca acatcgagtg ggctcccatt
     4741 aaagctgcac aacttactag accgaacgct gagttggttg aactcgaaat tgatacccta
     4801 catggtgtga aaacgatcag gtgtaccccc gaccacccag tttacactaa aaacaggggc
     4861 tacgtgcgag cggatgagtt aaccgatgat gatgagcttg ttgtagccat cgagaacctg
     4921 tatggcgatg aagtagatgg ctattttcag ggcatggaag cggagacgta tataggaaag
     4981 cttaaatccc gagcaattgt atcaaacgag gacacgtatg atattcagac gtcaactcat
     5041 aacttttttg cgaatgataa cagcgccggc ggagtgcagg tggaaaccat ctccccagga
     5101 gacgggcgca ccttccccaa gcgcggccag acctgcgtgg tgcactacac cgggatgctt
     5161 gaagatggaa agaaatttga ttcctcccgg gacagaaaca agccctttaa gtttatgcta
     5221 ggcaagcagg aggtgatccg aggctgggaa gaaggggttg cccagatgag tgtgggtcag
     5281 agagccaaac tgactatatc tccagattat gcctatggtg ccactgggca cccaggcatc
     5341 atcccaccac atgccactct cgtcttcgat gtggagcttc taaaactgga aggaagcggg
     5401 agtgggagca aaaagaagaa aaagaagtca aagacaaagt gtgtaattat gtagcctgca
     5461 ggcaattcgt cgagggacct aataacttcg tatagcatac attatacgaa gttatacatg
     5521 tttaagggtt ccggttccac taggtacaat tcgatatcaa gcttatcgat aatcaacctc
     5581 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     5641 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     5701 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     5761 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     5821 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     5881 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     5941 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     6001 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     6061 accttccttc ccggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     6121 cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg acctcgatcg
     6181 agacctagaa aaacatggag caatcacaag tagcaataca gcagctacca atgctgattg
     6241 tgcctggcta gaagcacaag aggaggagga ggtgggtttt ccagtcacac ctcaggtacc
     6301 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     6361 gggactggaa gggctaattc actcccaacg aagacaagat atccttgatc tgtggatcta
     6421 ccacacacaa ggctacttcc ctgattggca gaactacaca ccagggccag ggatcagata
     6481 tccactgacc tttggatggt gctacaagct agtaccagtt gagcaagaga aggtagaaga
     6541 agccaatgaa ggagagaaca cccgcttgtt acaccctgtg agcctgcatg ggatggatga
     6601 cccggagaga gaagtattag agtggaggtt tgacagccgc ctagcatttc atcacatggc
     6661 ccgagagctg catccggact gtactgggtc tctctggtta gaccagatct gagcctggga
     6721 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
     6781 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
     6841 ttagtcagtg tggaaaatct ctagcagcat gtgagcaaaa ggccagcaaa aggccaggaa
     6901 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6961 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7021 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     7081 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     7141 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     7201 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     7261 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7321 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     7381 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7441 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7501 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7561 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7621 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaactcggtc
     7681 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7741 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7801 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7861 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7921 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7981 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8041 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8101 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8161 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8221 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8281 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8341 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8401 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8461 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8521 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     8581 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     8641 aggggttccg cgcacatttc cccgaaaagt gccacctgac
//
