LOCUS       sequence-485467-        8546 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 8546)
  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 8546)
  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: pCMV5-BCRn-CAGEn (Npu)
FEATURES             Location/Qualifiers
     source          1..8546
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        178..557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        558..761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     712..732
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     833..849
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     833..849
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             3959..3979
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3980..4000
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             4220..4540
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             4619..4639
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4706..4732
                     /codon_start=1
                     /product="nuclear localization signal of human c-Myc 
                     proto-oncogene (Dang and Lee, 1988)"
                     /label=c-myc NLS
                     /translation="PAAKRVKLD"
     polyA_signal    4747..5369
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4946..4965)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(5393..5413)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5398..5727
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5578..5713
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5765..5784)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5766..5784)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5798..5814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5798..5814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5811..5833)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5822..5838
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5846..5852,5853..5870,5871..5876))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5891..5912
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6029..6046)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6200..6788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6280..6299)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6959..7750,7751..7819))
                     /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     7582..7601
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7820..7924)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7951..8406
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8038..8057)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8248..8269
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(8532..8546,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(8546,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg caattagcca
       61 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
      121 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
      181 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      421 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      481 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
      541 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
      601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      661 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
      721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
      781 attcagatct ggtaccacgc gtatcgataa gcttgcatgc ctgcaggtcg actctagaac
      841 tagtggatcc gttgtgatgg tggacccggt gggcttcgcg gaggcgtgga aggcgcagtt
      901 cccggactca gagcccccgc gcatggagct gcgctcagtg ggcgacatcg agcaggagct
      961 ggagcgctgc aaggcctcca ttcggcgcct ggagcaggag gtgaaccagg agcgcttccg
     1021 catgatctac ctgcagacgt tgctggccaa ggaaaagaag agctatgacc ggcagcgatg
     1081 gggcttccgg cgcgcggcgc aggcccccga cggcgcctcc gagccccgag cgtccgcgtc
     1141 gcgcccgcag ccagcgcccg ccgacggagc cgacccgccg cccgccgagg agcccgaggc
     1201 ccggcccgac ggcgagggtt ctccgggtaa ggccaggccc gggaccgccc gcaggcccgg
     1261 ggcagccgcg tcgggggaac gggacgaccg gggacccccc gccagcgtgg cggcgctcag
     1321 gtccaacttc gagcggatac gcaagggcca tggccagccc ggggcggacg ccgagaagcc
     1381 cttctacgtg aacgtcgagt ttcaccacga gcgcggcctg gtgaaggtca acgacaaaga
     1441 ggtgtcggac cgcatcagct ccctgggcag ccaggccatg cagatggagc gcaaaaagtc
     1501 ccagcacggc gcgggctcga gcgtggggga tgcatccagg cccccttacc ggggacgctc
     1561 ctcggagagc agctgcggcg tcgacggcga ctacgaggac gccgagttga acccccgctt
     1621 cctgaaggac aacctgatcg acgccaatgg cggtagcagg cccccttggc cgcccctgga
     1681 gtaccagccc taccagagca tctacgtcgg gggcatgatg gaaggggagg gcaagggccc
     1741 gctcctgcgc agccagagca cctctgagca ggagaagcgc cttacctggc cccgcaggtc
     1801 ctactccccc cggagttttg aggattgcgg aggcggctat accccggact gcagctccaa
     1861 tgagaacctc acctccagcg aggaggactt ctcctctggc cagtccagcc gcgtgtcccc
     1921 aagccccacc acctaccgca tgttccggga caaaagccgc tctccctcgc agaactcgca
     1981 acagtccttc gacagcagca gtccccccac gccgcagtgc cataagcggc accggcactg
     2041 cccggttgtc gtgtccgagg ccaccatcgt gggcgtccgc aagaccgggc agatctggcc
     2101 caacgatggc gagggcgcct tccatggaga cgcagatggc tcgttcggaa caccacctgg
     2161 atacggctgc gctgcagacc gggcagagga gcagcgccgg caccaagatg ggctgcccta
     2221 cattgatgac tcgccctcct catcgcccca cctcagcagc aagggcaggg gcagccggga
     2281 tgcgctggtc tcgggagccc tggagtccac taaagcgagt gagctggact tggaaaaggg
     2341 cttggagatg agaaaatggg tcctgtcggg aatcctggct agcgaggaga cttacctgag
     2401 ccacctggag gcactgctgc tgcccatgaa gcctttgaaa gccgctgcca ccacctctca
     2461 gccggtgctg acgagtcagc agatcgagac catcttcttc aaagtgcctg agctctacga
     2521 gatccacaag gagttctatg atgggctctt cccccgcgtg cagcagtgga gccaccagca
     2581 gcgggtgggc gacctcttcc agaagctggc cagccagctg ggtgtgtacc gggccttcgt
     2641 ggacaactac ggagttgcca tggaaatggc tgagaagtgc tgtcaggcca atgctcagtt
     2701 tgcagaaatc tccgagaacc tgagagccag aagcaacaaa gatgccaagg atccaacgac
     2761 caagaactct ctggaaactc tgctctacaa gcctgtggac cgtgtgacga ggagcacgct
     2821 ggtcctccat gacttgctga agcacactcc tgccagccac cctgaccacc ccttgctgca
     2881 ggacgccctc cgcatctcac agaacttcct gtccagcatc aatgaggaga tcacaccccg
     2941 acggcagtcc atgacggtga agaagggaga gcaccggcag ctgctgaagg acagcttcat
     3001 ggtggagctg gtggaggggg cccgcaagct gcgccacgtc ttcctgttca ccgacctgct
     3061 tctctgcacc aagctcaaga agcagagcgg aggcaaaacg cagcagtatg actgcaaatg
     3121 gtacattccg ctcacggatc tcagcttcca gatggtggat gaactggagg cagtgcccaa
     3181 catccccctg gtgcccgatg aggagctgga cgctttgaag atcaagatct cccagatcaa
     3241 gagtgacatc cagagagaga agagggcgaa caagggcagc aaggctacgg agaggctgaa
     3301 gaagaagctg tcggagcagg agtcactgct gctgcttatg tctcccagca tggccttcag
     3361 ggtgcacagc cgcaacggca agagttacac gttcctgatc tcctctgact atgagcgtgc
     3421 agagtggagg gagaacatcc gggagcagca gaagaagtgt ttcagaagct tctccctgac
     3481 atccgtggag ctgcagatgc tgaccaactc gtgtgtgaaa ctccagactg tccacagcat
     3541 tccgctgacc atcaataagg aagatgatga gtctccgggg ctctatgggt ttctgaatgt
     3601 catcgtccac tcagccactg gatttaagca gagttcaaaa tgtttaagct atgaaacgga
     3661 aatattgaca gtagaatatg gattattacc gattggtaaa attgtagaaa agcgcatcga
     3721 atgtactgtt tatagcgttg ataataatgg aaatatttat acacaacctg tagcacaatg
     3781 gcacgatcgc ggaaagcaaa aggtgtttga gtattgtttg gaggatggtt cattgattcg
     3841 ggcaacaaaa gaccataagt ttatgactgt tgatggacaa atgttgccaa ttaaggaaat
     3901 atttcgtcgt aagttggatt tgatgcgggt tgataatttg ccgaatggaa gcggaggaga
     3961 aaatctatac ttccaaggtg agaacctgta ttttcagggc gggagtggtg ggatcgaaat
     4021 agccacagaa aaatatttag gcgaacaaaa tgtctatgac attggagttg agcgcgacca
     4081 taattttgca ctcaaaaatg gcggctattt tcagggcatc gaaatagcca cagaaaaata
     4141 tttaggcgaa caaaatgtct atgacattgg agttgagcgc gaccataatt ttgcactcaa
     4201 aaatggcaac agcgccggcg gagtgcaggt ggaaaccatc tccccaggag acgggcgcac
     4261 cttccccaag cgcggccaga cctgcgtggt gcactacacc gggatgcttg aagatggaaa
     4321 gaaatttgat tcctcccggg acagaaacaa gccctttaag tttatgctag gcaagcagga
     4381 ggtgatccga ggctgggaag aaggggttgc ccagatgagt gtgggtcaga gagccaaact
     4441 gactatatct ccagattatg cctatggtgc cactgggcac ccaggcatca tcccaccaca
     4501 tgccactctc gtcttcgatg tggagcttct aaaactggaa gaacagaaac tgattagtga
     4561 ggaagatgga ggaagtggag gtagtgcggc cgctccaatt gccggatcca aggctagccc
     4621 gaaaaagaaa cgcaaagttg ggcgcggttc tggttcagct gctgccaaga agaagaagtt
     4681 ggactcaggg tcaggttctt caggtcctgc tgccaagagg gtcaagttgg acgcctagag
     4741 gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     4801 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     4861 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     4921 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     4981 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     5041 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     5101 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     5161 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     5221 cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca taggctacct
     5281 ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt
     5341 gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg
     5401 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     5461 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5521 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5581 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5641 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5701 ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt
     5761 aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa
     5821 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     5881 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     5941 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     6001 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6061 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     6361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     6421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     6481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     6541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     6601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     6661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     6721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     6781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     6841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     6901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     6961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     7021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     7081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     7141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     7201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     7261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     7321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     7381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     7441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     7501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     7561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     7621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     7681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     7741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     7801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     7861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     7921 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     7981 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     8041 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     8101 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     8161 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     8221 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     8281 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     8341 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     8401 caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     8461 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     8521 gccagggttt tcccagtcac gacgtt
//
