LOCUS       sequence-485496-        9414 bp    DNA     circular SYN 08-SEP-2025
DEFINITION  Proximity-triggered Protein Trans-Splicing to generate the splice 
            product BCR-ABL, Figure 3, Extended Figure 6-7 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 9414)
  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 9414)
  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-CAGEc-ABL1 (Npu)
FEATURES             Location/Qualifiers
     source          1..9414
                     /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             1840..2115
                     /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             2299..2319
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2320..2340
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             5776..5802
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    6276..6397
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6313..6332)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6367..6386
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6431..6448)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6602..7190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6682..6701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7361..8152,8153..8221))
                     /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     7984..8003
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8222..8326)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8394..8412
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8450..8472)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8572..8591
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8658..9113)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8795..8816)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     9007..9026
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9239..9261
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9253..9270
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9254..9270
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9307..9327)
                     /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 gcgtttcatt
     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 ggggcagcag cctggaaaag tacttgggga ccaaagaagg
     1561 ccaagcttgc ctgccctgca ttttatcaaa ggagcaggga agaaggaatc atcgaggcat
     1621 gggggtccac actgcaatgt ttttgtggaa catgaagccc ttcagcggcc agtagcatct
     1681 gactttgagc ctcagggtct gagtgaagcc gctcgttgga actccaagga aaaccttctc
     1741 gctggaccca gtgaaggagg aagtggaggt agtggtggag gaggctctgg tggaggcggt
     1801 agtggttctg gttcaggaag tggtagcgga agagtagcga tcctctggca tgagatgtgg
     1861 catgaaggcc tggaagaggc atctcgtttg tactttgggg aaaggaacgt gaaaggcatg
     1921 tttgaggtgc tggagccctt gcatgctatg atggaacggg gcccccagac tctgaaggaa
     1981 acatccttta atcaggccta tggtcgagat ttaatggagg cccaagagtg gtgcaggaag
     2041 tacatgaaat cagggaatgt caaggacctc ctccaagcct gggacctcta ttatcatgtg
     2101 ttccgacgaa tctcaggaaa tggaaatgga ggagaacaag aggtgtttga gtattgtttg
     2161 gaagatggtt cattgattcg ggcaacaaaa gaccataagt ttatgactgt tgatggtcaa
     2221 atgttgccaa ttgatgaaat atttgaacgt gaattggatt tgatgcgggt tgataatttg
     2281 ccgaatggaa gtggaggaga gaacctgtat tttcagggcg agaacctgta ttttcagggc
     2341 ggaagtggag gaatcaaaat agccacacgt aaatatttag gcaaacaaaa tgtctatgac
     2401 attggagttg agcgcgacca taattttgca ctcaaaaatg gcttcatagc ttctaattgc
     2461 tttaatgccc ttcagcggcc agtagcatct gactttgagc ctcagggtct gagtgaagcc
     2521 gctcgttgga actccaagga aaaccttctc gctggaccca gtgaaaatga ccccaacctt
     2581 ttcgttgcac tgtatgattt tgtggccagt ggagataaca ctctaagcat aactaaaggt
     2641 gaaaagctcc gggtcttagg ctataatcac aatggggaat ggtgtgaagc ccaaaccaaa
     2701 aatggccaag gctgggtccc aagcaactac atcacgccag tcaacagtct ggagaaacac
     2761 tcctggtacc atgggcctgt gtcccgcaat gccgctgagt atctgctgag cagcgggatc
     2821 aatggcagct tcttggtgcg tgagagtgag agcagtcctg gccagaggtc catctcgctg
     2881 agatacgaag ggagggtgta ccattacagg atcaacactg cttctgatgg caagctctac
     2941 gtctcctccg agagccgctt caacaccctg gccgagttgg ttcatcatca ttcaacggtg
     3001 gccgacgggc tcatcaccac gctccattat ccagccccaa agcgcaacaa gcccactgtc
     3061 tatggtgtgt cccccaacta cgacaagtgg gagatggaac gcacggacat caccatgaag
     3121 cacaagctgg gcgggggcca gtacggggag gtgtacgagg gcgtgtggaa gaaatacagc
     3181 ctgacggtgg ccgtgaagac cttgaaggag gacaccatgg aggtggaaga gttcttgaaa
     3241 gaagctgcag tcatgaaaga gatcaaacac cctaacctgg tgcagctcct tggggtctgc
     3301 acccgggagc ccccgttcta tatcatcact gagttcatga cctacgggaa cctcctggac
     3361 tacctgaggg agtgcaaccg gcaggaggtg aacgccgtgg tgctgctgta catggccact
     3421 cagatctcgt cagccatgga gtacctggag aagaaaaact tcatccacag agatcttgct
     3481 gcccgaaact gcctggtagg ggagaaccac ttggtgaagg tagctgattt tggcctgagc
     3541 aggttgatga caggggacac ctacacagcc catgctggag ccaagttccc catcaaatgg
     3601 actgcacccg agagcctggc ctacaacaag ttctccatca agtccgacgt ctgggcattt
     3661 ggagtattgc tttgggaaat tgctacctat ggcatgtccc cttacccggg aattgacctg
     3721 tcccaggtgt atgagctgct agagaaggac taccgcatgg agcgcccaga aggctgccca
     3781 gagaaggtct atgaactcat gcgagcatgt tggcagtgga atccctctga ccggccctcc
     3841 tttgctgaaa tccaccaagc ctttgaaaca atgttccagg aatccagtat ctcagacgaa
     3901 gtggaaaagg agctggggaa acaaggcgtc cgtggggctg tgagtacctt gctgcaggcc
     3961 ccagagctgc ccaccaagac gaggacctcc aggagagctg cagagcacag agacaccact
     4021 gacgtgcctg agatgcctca ctccaagggc cagggagaga gcgatcctct ggaccatgag
     4081 cctgccgtgt ctccattgct ccctcgaaaa gagcgaggtc ccccggaggg cggcctgaat
     4141 gaagatgagc gccttctccc caaagacaaa aagaccaact tgttcagcgc cttgatcaag
     4201 aagaagaaga agacagcccc aacccctccc aaacgcagca gctccttccg ggagatggac
     4261 ggccagccgg agcgcagagg ggccggcgag gaagagggcc gagacatcag caacggggca
     4321 ctggctttca cccccttgga cacagctgac ccagccaagt ccccaaagcc cagcaatggg
     4381 gctggggtcc ccaatggagc cctccgggag tccgggggct caggcttccg gtctccccac
     4441 ctgtggaaga agtccagcac gctgaccagc agccgcctag ccaccggcga ggaggagggc
     4501 ggtggcagct ccagcaagcg cttcctgcgc tcttgctccg cctcctgcgt tccccatggg
     4561 gccaaggaca cggagtggag gtcagtcacg ctgcctcggg acttgcagtc cacgggaaga
     4621 cagtttgact cgtccacatt tggagggcac aaaagtgaga agccggctct gcctcggaag
     4681 agggcagggg agaacaggtc tgaccaggtg acccgaggca cagtaacgcc tccccccagg
     4741 ctggtgaaaa agaatgagga agctgctgat gaggtcttca aagacatcat ggagtccagc
     4801 ccgggctcca gcccgcccaa cctgactcca aaacccctcc ggcggcaggt caccgtggcc
     4861 cctgcctcgg gcctccccca caaggaagaa gctggaaagg gcagtgcctt agggacccct
     4921 gctgcagctg agccagtgac ccccaccagc aaagcaggct caggtgcacc agggggcacc
     4981 agcaagggcc ccgccgagga gtccagagtg aggaggcaca agcactcctc tgagtcgcca
     5041 gggagggaca aggggaaatt gtccaggctc aaacctgccc cgccgccccc accagcagcc
     5101 tctgcaggga aggctggagg aaagccctcg cagagcccga gccaggaggc ggccggggag
     5161 gcagtcctgg gcgcaaagac aaaagccacg agtctggttg atgctgtgaa cagtgacgct
     5221 gccaagccca gccagccggg agagggcctc aaaaagcccg tgctcccggc cactccaaag
     5281 ccacagtccg ccaagccgtc ggggaccccc atcagcccag cccccgttcc ctccacgttg
     5341 ccatcagcat cctcggccct ggcaggggac cagccgtctt ccaccgcctt catccctctc
     5401 atatcaaccc gagtgtctct tcggaaaacc cgccagcctc cagagcggat cgccagcggc
     5461 gccatcacca agggcgtggt cctggacagc accgaggcgc tgtgcctcgc catctctagg
     5521 aactccgagc agatggccag ccacagcgca gtgctggagg ccggcaaaaa cctctacacg
     5581 ttctgcgtga gctatgtgga ttcaatccag caaatgagga acaagtttgc cttccgagag
     5641 gccatcaaca aactggagaa taatctccgg gagcttcaga tctgcccggc gacagcaggc
     5701 agtggtccag cggccactca ggacttcagc aagctcctca gttcggtgaa ggaaatcagt
     5761 gacatagtgc agaggtaccc gtacgacgtt ccggactacg cttcttagca gcagtcaggg
     5821 gtcaggtgtc aggcccgtcg gagctgcctg cagcacatgc gggctcgccc atacccgtga
     5881 cagtggctga caagggacta gtgagtcagc accttggccc aggagctctg cgccaggcag
     5941 agctgagggc cctgtggagt ccagctctac tacctacgtt tgcaccgcct gccctcccgc
     6001 accttcctcc tccccgctcc gtctctgtcc tcgaatttta tctgtggagt tcctgctccg
     6061 tggactgcag tcggcatgcc aggacccgcc agccccgctc ccacctagtg ccccagactg
     6121 agctctccag gccaggtggg aacggctgat gtggactgtc tttttcattt ttttctctct
     6181 ggagcccctc ctcccccggc tgggcctcct tcttccactt ctccaagaat ggaagcctga
     6241 actgaggaat tcggatccag atcttattaa agcagaactt gtttattgca gcttataatg
     6301 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     6361 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggtcg actctagact
     6421 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6481 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6541 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6601 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6661 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6721 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6781 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6841 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6901 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6961 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7021 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7081 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7141 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7201 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7261 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7321 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7381 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7441 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7501 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7561 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7621 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7681 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7741 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7801 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7861 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7921 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7981 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8041 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8101 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8161 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8221 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8281 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8341 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     8401 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     8461 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     8521 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     8581 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     8641 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     8701 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     8761 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     8821 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     8881 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     8941 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     9001 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     9061 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     9121 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     9181 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     9241 cagtcacgac gttgtaaaac gacggccagt gaattgtcga cttctgaggc ggaaagaacc
     9301 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     9361 gtatgcaaag catgcatctc aattagtcag caaggaaagt ccccaggctc ccca
//
