LOCUS       sequence-488594-        6894 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Proximity-triggered Protein Trans-Splicing to generate the splice 
            product DNAJ-PKAc, Figure 4, Extended Figure 8-9 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 6894)
  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 6894)
  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: pSG5-CAGEc-PRKACA (Nrdj1)
FEATURES             Location/Qualifiers
     source          1..6894
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..211
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        23..219
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      70..205
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     intron          250..822
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     830..849
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     877..896
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        877..895
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1717..1992
                     /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             3256..3282
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3756..3877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3793..3812)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3847..3866
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3911..3928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4082..4670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4162..4181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4841..5632,5633..5701))
                     /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     5464..5483
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5702..5806)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5874..5892
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5930..5952)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6052..6071
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6138..6593)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6275..6296)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6487..6506
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6719..6741
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6733..6750
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6734..6750
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6787..6807)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
       61 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
      121 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
      181 tagtgaagag gcttttttgg aggcctaggc ttttgcaaaa agctccggat cgatcctgag
      241 aacttcaggg tgagtttggg gacccttgat tgttctttct ttttcgctat tgtaaaattc
      301 atgttatatg gagggggcaa agttttcagg gtgttgttta gaatgggaag atgtcccttg
      361 tatcaccatg gaccctcatg ataattttgt ttctttcact ttctactctg ttgacaacca
      421 ttgtctcctc ttattttctt ttcattttct gtaacttttt cgttaaactt tagcttgcat
      481 ttgtaacgaa tttttaaatt cacttttgtt tatttgtcag attgtaagta ctttctctaa
      541 tcactttttt ttcaaggcaa tcagggtata ttatattgta cttcagcaca gttttagaga
      601 acaattgtta taattaaatg ataaggtaga atatttctgc atataaattc tggctggcgt
      661 ggaaatattc ttattggtag aaacaactac atcctggtca tcatcctgcc tttctcttta
      721 tggttacaat gatatacact gtttgagatg aggataaaat actctgagtc caaaccgggc
      781 ccctctgcta accatgttca tgccttcttc tttttcctac agctcctggg caacgtgctg
      841 gttattgtgc tgtctcatca ttttggtaaa gaattgtaat acgactcact atagggcgaa
      901 ttccttttaa aaagcggact taaaagttgc tggcaatgca ttccttttcg tcagagtcga
      961 gggcaaactc gctgaaatct gggtgacccg tgtccttttc cggagagcaa agcagagaag
     1021 cgagagcggc cactagttcg gcaggaaatt tgttggaaga tgaagaagct aagatagggg
     1081 gttggtgact tccacaggaa aagttctgga ggagtagcca aagaccatca gcgtttcctt
     1141 tatgtgtgag aattgaaatg actagcatta ttgacccttt tcagcatccc ctgtgaatat
     1201 ttctgtttag gtttttcttc ttgaaaagaa attgttattc agcccgttta aaacaaatca
     1261 agaaactttt gggtaacatt gcaattacat gaaattgata accgcgaaaa taattggaac
     1321 tcctgcttgc aagtgtcaac ctaaaaaaag tgcttccttt tgttatggaa gatgtctttc
     1381 tgtgattgac ttcaattgct gacttgtgga gatgcagcga atgtgaaatc ccacgtatat
     1441 gccatttccc tctacgctcg ctgaccgttc tggaagatct tgaaccctct tctggaaagg
     1501 ggtacctatt attactttat gggcaacgcc gccgccgcca agaagggcag cgagcaggag
     1561 agcgtgaaag aattcttagc caaagccaaa gaagattttc ttaaaaaatg ggaaagtccc
     1621 gctcagaaca caggaggaag tggaggtagt ggtggaggag gctctggtgg aggcggtagt
     1681 ggttctggtt caggaagtgg tagcggaaga gtagcgatcc tctggcatga gatgtggcat
     1741 gaaggcctgg aagaggcatc tcgtttgtac tttggggaaa ggaacgtgaa aggcatgttt
     1801 gaggtgctgg agcccttgca tgctatgatg gaacggggcc cccagactct gaaggaaaca
     1861 tcctttaatc aggcctatgg tcgagattta atggaggccc aagagtggtg caggaagtac
     1921 atgaaatcag ggaatgtcaa ggacctcctc caagcctggg acctctatta tcatgtgttc
     1981 cgacgaatct caggaaatgg aaatggaaac gctgagttgg ttgaactcga aattaagacc
     2041 ctacatggtg tgaaaacgat caggtgtacc cccgaccacc cagtttacac taaaaacagg
     2101 ggctacgtgc gagcggatga gttaaccgat gatgatgagc ttgttgtagc catcgagaac
     2161 ctgtatggcg atgaagtaga tggctatttt cagggcatgg aagcgaagac gtatatagga
     2221 aagcttaaat cccgaaaaat tgtatcaaac gaggacacgt atgatattca gacgtcaact
     2281 cataactttt ttgcgaatga tatacttgtt cataatagtc acttggatca gtttgaacga
     2341 atcaagaccc tcggcacggg ctccttcggg cgggtgatgc tggtgaaaca caaggagacc
     2401 gggaaccact atgccatgaa gatcctcgac aaacagaagg tggtgaaact gaaacagatc
     2461 gaacacaccc tgaatgaaaa gcgcatcctg caagctgtca actttccgtt cctcgtcaaa
     2521 ctcgagttct ccttcaagga caactcaaac ttatacatgg tcatggagta cgtgcccggc
     2581 ggggagatgt tctcacacct acggcggatc ggaaggttca gtgagcccca tgcccgtttc
     2641 tacgcggccc agatcgtcct gacctttgag tatctgcact cgctggatct catctacagg
     2701 gacctgaagc cggagaatct gctcattgac cagcagggct acattcaggt gacagacttc
     2761 ggtttcgcca agcgcgtgaa gggccgcact tggaccttgt gcggcacccc tgagtacctg
     2821 gcccctgaga ttatcctgag caaaggctac aacaaggccg tggactggtg ggccctgggg
     2881 gttcttatct atgaaatggc cgctggctac ccgcccttct tcgcagacca gcccatccag
     2941 atctatgaga agatcgtctc tgggaaggtg cgcttccctt cccacttcag ctctgacttg
     3001 aaggacctgc tgcggaacct cctgcaggta gatctcacca agcgctttgg gaacctcaag
     3061 aatggggtca acgatatcaa gaaccacaag tggtttgcca caactgactg gattgccatc
     3121 taccagagga aggtggaagc tcccttcata ccaaagttta aaggccctgg ggatacgagt
     3181 aactttgacg actatgagga agaagaaatc cgggtctcca tcaatgagaa gtgtggcaag
     3241 gagttttctg agttttaccc gtacgacgtt ccggactacg cttcttagca gcagtcaggg
     3301 gtcaggtgtc aggcccgtcg gagctgcctg cagcacatgc gggctcgccc atacccgtga
     3361 cagtggctga caagggacta gtgagtcagc accttggccc aggagctctg cgccaggcag
     3421 agctgagggc cctgtggagt ccagctctac tacctacgtt tgcaccgcct gccctcccgc
     3481 accttcctcc tccccgctcc gtctctgtcc tcgaatttta tctgtggagt tcctgctccg
     3541 tggactgcag tcggcatgcc aggacccgcc agccccgctc ccacctagtg ccccagactg
     3601 agctctccag gccaggtggg aacggctgat gtggactgtc tttttcattt ttttctctct
     3661 ggagcccctc ctcccccggc tgggcctcct tcttccactt ctccaagaat ggaagcctga
     3721 actgaggaat tcggatccag atcttattaa agcagaactt gtttattgca gcttataatg
     3781 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3841 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggtcg actctagact
     3901 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3961 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4021 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4081 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4141 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4201 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4261 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4321 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4381 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4441 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4501 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4561 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4621 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4681 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4741 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4801 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4861 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4921 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4981 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5041 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5101 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5161 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5221 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5281 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5341 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5401 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5461 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5521 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5581 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5641 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5701 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5761 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5821 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5881 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     5941 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     6001 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     6061 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     6121 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     6181 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     6241 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     6301 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     6361 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     6421 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     6481 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     6541 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     6601 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     6661 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     6721 cagtcacgac gttgtaaaac gacggccagt gaattgtcga cttctgaggc ggaaagaacc
     6781 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     6841 gtatgcaaag catgcatctc aattagtcag caaggaaagt ccccaggctc ccca
//
