LOCUS       sequence-485476-        6288 bp    DNA     circular SYN 08-SEP-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 6288)
  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 6288)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 8, 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-DNAJB1n_CAGEn (Nrdj1)
FEATURES             Location/Qualifiers
     source          1..6288
                     /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             2272..2592
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3150..3271
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3187..3206)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3241..3260
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3305..3322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3476..4064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3556..3575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4235..5026,5027..5095))
                     /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     4858..4877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5096..5200)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5268..5286
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5324..5346)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5446..5465
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5532..5987)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5669..5690)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5881..5900
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6113..6135
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6127..6144
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6128..6144
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6181..6201)
                     /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 gggtaaagac tactaccaga cgttgggcct ggcccgcggc
     1561 gcgtcggacg aggagatcaa gcgggcctac cgccgccagg cgctgcgcta ccacccggac
     1621 aagaacaagg agcccggcgc cgaggagaag ttcaaggaga tcgctgaggc ctacgacgtg
     1681 ctcagcgacc cgcgcaagcg cgagatcttc gaccgctacg gggaggaagt gaaagaattc
     1741 ttagccaaag ccaaagaaga ttttcttaaa aaatgggaaa gtcccgctca gaacacatgt
     1801 ctggtcggct ctagcgaaat catcacacgt aattatggga agacaaccat taaggaagta
     1861 gttgagatat ttgacaatga taaaaatatc caagttctag cgtttaacac tcacactgac
     1921 aacatcgagt gggctcccat taaagctgca caacttacta gaccgaacgc tgagttggtt
     1981 gaactcgaaa ttgataccct acatggtgtg aaaacgatca ggtgtacccc cgaccaccca
     2041 gtttacacta aaaacagggg ctacgtgcga gcggatgagt taaccgatga tgatgagctt
     2101 gttgtagcca tcgagaacct gtatggcgat gaagtagatg gctattttca gggcatggaa
     2161 gcggagacgt atataggaaa gcttaaatcc cgagcaattg tatcaaacga ggacacgtat
     2221 gatattcaga cgtcaactca taactttttt gcgaatgata acagcgccgg cggagtgcag
     2281 gtggaaacca tctccccagg agacgggcgc accttcccca agcgcggcca gacctgcgtg
     2341 gtgcactaca ccgggatgct tgaagatgga aagaaatttg attcctcccg ggacagaaac
     2401 aagcccttta agtttatgct aggcaagcag gaggtgatcc gaggctggga agaaggggtt
     2461 gcccagatga gtgtgggtca gagagccaaa ctgactatat ctccagatta tgcctatggt
     2521 gccactgggc acccaggcat catcccacca catgccactc tcgtcttcga tgtggagctt
     2581 ctaaaactgg aagaacagaa actgattagt gaggaagatg gaggaagtgg aggtagtgct
     2641 ctccaaaaaa agttggaaga gcttgaactg gacgaacagt agcagcagtc aggggtcagg
     2701 tgtcaggccc gtcggagctg cctgcagcac atgcgggctc gcccataccc gtgacagtgg
     2761 ctgacaaggg actagtgagt cagcaccttg gcccaggagc tctgcgccag gcagagctga
     2821 gggccctgtg gagtccagct ctactaccta cgtttgcacc gcctgccctc ccgcaccttc
     2881 ctcctccccg ctccgtctct gtcctcgaat tttatctgtg gagttcctgc tccgtggact
     2941 gcagtcggca tgccaggacc cgccagcccc gctcccacct agtgccccag actgagctct
     3001 ccaggccagg tgggaacggc tgatgtggac tgtctttttc atttttttct ctctggagcc
     3061 cctcctcccc cggctgggcc tccttcttcc acttctccaa gaatggaagc ctgaactgag
     3121 gaattcggat ccagatctta ttaaagcaga acttgtttat tgcagcttat aatggttaca
     3181 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3241 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gtcgactcta gactcttccg
     3301 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3361 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3421 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3481 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3541 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3601 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3661 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3721 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3781 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3841 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3901 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3961 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4021 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4081 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4141 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4201 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4261 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4321 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4381 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4441 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4501 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4561 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4621 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4681 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4741 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4801 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4861 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4921 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4981 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5041 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5101 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5161 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5221 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     5281 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     5341 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     5401 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     5461 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     5521 accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     5581 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     5641 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     5701 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     5761 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     5821 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     5881 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     5941 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc
     6001 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     6061 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     6121 cgacgttgta aaacgacggc cagtgaattg tcgacttctg aggcggaaag aaccagctgt
     6181 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6241 aaagcatgca tctcaattag tcagcaagga aagtccccag gctcccca
//
