LOCUS       sequence-493945-        5782 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 5782)
  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 5782)
  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-mVenus-CAGEn (Nrdj1)
FEATURES             Location/Qualifiers
     source          1..5782
                     /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      892..901
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             928..951
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             961..984
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             994..1017
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(1033..1035,1036..1038,1039..1749)
                     /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(1078..1099)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1339..1358)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1686..1707
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2233..2556
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     protein_bind    complement(2584..2617)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2673..3259
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2726..2746)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3144..3155)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3262..3278)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3263..3279)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3788..3968
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4030..4618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4110..4129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4789..5580,5581..5649))
                     /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     5412..5431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5650..5754)
                     /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 ttctgcagtc gacggtaccg cgggcccggg atccaccggt cgccaccatg
      901 ggtagtctcg agggtaccat gggttcagac tataaagatg atgatgataa aggtggctcc
      961 gactacaagg atgacgatga caagggaggt agtgactaca aagacgatga cgacaagggc
     1021 ggaggaggaa gtatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1081 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1141 gatgccacct acggcaagct gaccctgaag ttgatctgca ccaccggcaa gctgcccgtg
     1201 ccctggccca ccctcgtgac caccctcggc tacggcctgc agtgcttcgc ccgctacccc
     1261 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1321 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgat
     1381 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1441 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
     1501 aagcagaaga acggcatcaa ggcaaacttc aagatccgcc acaacatcga ggacggcggc
     1561 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1621 cccgacaacc actacctgag ctaccagtcc aaactgagca aagaccccaa cgagaagcgc
     1681 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1741 ctgtacaaga atccatgctg tctggtcggc tctagcgaaa tcatcacacg taattatggg
     1801 aagacaacca ttaaggaagt agttgagata tttgacaatg ataaaaatat ccaagttcta
     1861 gcgtttaaca ctcacactga caacatcgag tgggctccca ttaaagctgc acaacttact
     1921 agaccgaacg ctgagttggt tgaactcgaa attgataccc tacatggtgt gaaaacgatc
     1981 aggtgtaccc ccgaccaccc agtttacact aaaaacaggg gctacgtgcg agcggatgag
     2041 ttaaccgatg atgatgagct tgttgtagcc atcgagaacc tgtatggcga tgaagtagat
     2101 ggctattttc agggcatgga agcggagacg tatataggaa agcttaaatc ccgagcaatt
     2161 gtatcaaacg aggacacgta tgatattcag acgtcaactc ataacttttt tgcgaatgat
     2221 aacagcgccg gcggagtgca ggtggaaacc atctccccag gagacgggcg caccttcccc
     2281 aagcgcggcc agacctgcgt ggtgcactac accgggatgc ttgaagatgg aaagaaattt
     2341 gattcctccc gggacagaaa caagcccttt aagtttatgc taggcaagca ggaggtgatc
     2401 cgaggctggg aagaaggggt tgcccagatg agtgtgggtc agagagccaa actgactata
     2461 tctccagatt atgcctatgg tgccactggg cacccaggca tcatcccacc acatgccact
     2521 ctcgtcttcg atgtggagct tctaaaactg gaatagcctg caggcaattc gtcgagggac
     2581 ctaataactt cgtatagcat acattatacg aagttataca tgtttaaggg ttccggttcc
     2641 actaggtaca attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     2701 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     2761 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     2821 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     2881 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     2941 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3001 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3061 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     3121 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccggcctg
     3181 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     3241 ctttgggccg cctccccgca tcgataccgt cgacctcgat cgagacctag aaaaacatgg
     3301 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca
     3361 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta
     3421 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     3481 tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt
     3541 ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga cctttggatg
     3601 gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa
     3661 cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtatt
     3721 agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc tgcatccgga
     3781 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     3841 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     3901 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     3961 ctctagcagc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4021 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4081 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4141 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4201 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4261 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4321 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4381 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4441 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4501 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4561 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4621 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4681 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4741 aagttttaaa tcaatctaaa gtatatatga gtaaactcgg tctgacagtt accaatgctt
     4801 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4861 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4921 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4981 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5041 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5101 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5161 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5221 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5281 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5341 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5401 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5461 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5521 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5581 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5641 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5701 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5761 tccccgaaaa gtgccacctg ac
//
