LOCUS       sequence-493941-        5499 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 5499)
  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 5499)
  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 (Nrdj1)
FEATURES             Location/Qualifiers
     source          1..5499
                     /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"
     protein_bind    complement(2301..2334)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2390..2976
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2443..2463)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2861..2872)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2979..2995)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2980..2996)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             3505..3685
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(3747..4335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3827..3846)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4506..5297,5298..5366))
                     /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     5129..5148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5367..5471)
                     /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 tacgcttctt aaggatccac cggatcaaga taacctgcag
     2281 gcaattcgtc gagggaccta ataacttcgt atagcataca ttatacgaag ttatacatgt
     2341 ttaagggttc cggttccact aggtacaatt cgatatcaag cttatcgata atcaacctct
     2401 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2461 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2521 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2581 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2641 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2701 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2761 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     2821 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2881 ccttccttcc cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     2941 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgatcga
     3001 gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt
     3061 gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct
     3121 ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg
     3181 ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac
     3241 cacacacaag gctacttccc tgattggcag aactacacac cagggccagg gatcagatat
     3301 ccactgacct ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa
     3361 gccaatgaag gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac
     3421 ccggagagag aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc
     3481 cgagagctgc atccggactg tactgggtct ctctggttag accagatctg agcctgggag
     3541 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     3601 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     3661 tagtcagtgt ggaaaatctc tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac
     3721 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3781 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3841 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3901 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3961 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4021 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4081 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4141 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4201 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4261 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4321 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4381 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4441 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4501 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4561 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4621 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     4681 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4741 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     4801 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     4861 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     4921 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     4981 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5041 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5101 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5161 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5221 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5281 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5341 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5401 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5461 ggggttccgc gcacatttcc ccgaaaagtg ccacctgac
//
