LOCUS       sequence-493943-        5658 bp    DNA     circular SYN 15-DEC-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 5658)
  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 5658)
  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: "bioRxiv 
            [Preprint]. 2023 Jul 10:2023.07.10.548208. doi: 
            10.1101/2023.07.10.548208."
COMMENT     Sequence Label: pLentiLox7.0-CAGEc-RFP-NLS (Nrdj1)
FEATURES             Location/Qualifiers
     source          1..5658
                     /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        618..821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      861..870
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             882..1157
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRIS"
     CDS             1491..2219
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="VSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMR
                     IKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGV
                     LTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTDM
                     ALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVA
                     RYCDLPSKLGHKLNGMDELYK"
     CDS             2220..2246
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2292..2312
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2379..2405
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     protein_bind    complement(2460..2493)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2549..3135
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2602..2622)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3020..3031)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3138..3154)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3139..3155)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3664..3844
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3906..4494)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3986..4005)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4665..5456,5457..5525))
                     /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     5288..5307
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5526..5630)
                     /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 taagcagagc tggtttagtg aaccgtcaga
      841 tccgctagaa ttgaccggtc gccaccatgg gaagagtagc gatcctctgg catgagatgt
      901 ggcatgaagg cctggaagag gcatctcgtt tgtactttgg ggaaaggaac gtgaaaggca
      961 tgtttgaggt gctggagccc ttgcatgcta tgatggaacg gggcccccag actctgaagg
     1021 aaacatcctt taatcaggcc tatggtcgag atttaatgga ggcccaagag tggtgcagga
     1081 agtacatgaa atcagggaat gtcaaggacc tcctccaagc ctgggacctc tattatcatg
     1141 tgttccgacg aatctcagga aatggaaatg gaaacgctga gttggttgaa ctcgaaatta
     1201 agaccctaca tggtgtgaaa acgatcaggt gtacccccga ccacccagtt tacactaaaa
     1261 acaggggcta cgtgcgagcg gatgagttaa ccgatgatga tgagcttgtt gtagccatcg
     1321 agaacctgta tggcgatgaa gtagatggct attttcaggg catggaagcg aagacgtata
     1381 taggaaagct taaatcccga aaaattgtat caaacgagga cacgtatgat attcagacgt
     1441 caactcataa cttttttgcg aatgatatac ttgttcataa tagtgagatc gtgtctaagg
     1501 gcgaagagct gattaaggag aacatgcaca tgaagctgta catggagggc accgtgaaca
     1561 accaccactt caagtgcaca tccgagggcg aaggcaagcc ctacgagggc acccagacca
     1621 tgagaatcaa ggtggtcgag ggcggccctc tccccttcgc cttcgacatc ctggctacca
     1681 gcttcatgta cggcagcaga accttcatca accacaccca gggcatcccc gacttcttta
     1741 agcagtcctt ccctgagggc ttcacatggg agagagtcac cacatacgaa gacgggggcg
     1801 tgctgaccgc tacccaggac accagcctcc aggacggctg cctcatctac aacgtcaaga
     1861 tcagaggggt gaacttccca tccaacggcc ctgtgatgca gaagaaaaca ctcggctggg
     1921 aggccaacac cgagatgctg taccccgctg acggcggcct ggaaggcaga accgacatgg
     1981 ccctgaagct cgtgggcggg ggccacctga tctgcaactt caagaccaca tacagatcca
     2041 agaaacccgc taagaacctc aagatgcccg gcgtctacta tgtggaccac agactggaaa
     2101 gaatcaagga ggccgacaaa gagacctacg tcgagcagca cgaggtggct gtggccagat
     2161 actgcgacct ccctagcaaa ctggggcaca aacttaatgg catggacgag ctgtacaagt
     2221 acccgtacga cgttccggac tacgcttctg gaggtagtgc ggccgctcca attgccggat
     2281 ccaaggctag cccgaaaaag aaacgcaaag ttgggcgcgg ttctggttca gctgctgcca
     2341 agaagaagaa gttggactca gggtcaggtt cttcaggtcc tgctgccaag agggtcaagt
     2401 tggacgccta aggatccacc ggatcaagat aacctgcagg caattcgtcg agggacctaa
     2461 taacttcgta tagcatacat tatacgaagt tatacatgtt taagggttcc ggttccacta
     2521 ggtacaattc gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag
     2581 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     2641 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     2701 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     2761 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     2821 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     2881 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     2941 gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     3001 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc ggcctgctgc
     3061 cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt
     3121 gggccgcctc cccgcatcga taccgtcgac ctcgatcgag acctagaaaa acatggagca
     3181 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
     3241 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
     3301 gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac
     3361 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     3421 gattggcaga actacacacc agggccaggg atcagatatc cactgacctt tggatggtgc
     3481 tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg agagaacacc
     3541 cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtattagag
     3601 tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca tccggactgt
     3661 actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac
     3721 ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg
     3781 ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct
     3841 agcagcatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3901 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3961 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4021 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4081 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4141 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4201 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4261 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4321 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4381 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4441 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4501 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4561 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4621 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4681 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4741 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4801 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     4861 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     4921 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     4981 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5041 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5101 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5161 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5221 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5281 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5341 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5401 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5461 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5521 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5581 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5641 cgaaaagtgc cacctgac
//
