LOCUS       sequence-485432-        6185 bp    DNA     circular SYN 05-SEP-2025
DEFINITION  Proximity-triggered Protein Trans-Splicing to generate the splice 
            product AML1-ETO, Extended Figure 3 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 6185)
  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 6185)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 5, 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-AMLn-CAGEn-NES (Npu)
FEATURES             Location/Qualifiers
     source          1..6185
                     /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             1706..1726
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1727..1747
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1967..2287
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    2386..3008
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2585..2604)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3032..3052)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3037..3366
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3217..3352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3404..3423)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3405..3423)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3437..3453)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3437..3453)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3450..3472)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3461..3477
                     /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(3485..3491,3492..3509,3510..3515))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3530..3551
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3668..3685)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3839..4427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3919..3938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4598..5389,5390..5458))
                     /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     5221..5240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5459..5563)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5590..6045
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5677..5696)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5887..5908
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(6171..6185,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6185,1..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 gttgtgatgc gtatccccgt agatgccagc acgagccgcc gcttcacgcc
      901 gccttccacc gcgctgagcc caggcaagat gagcgaggcg ttgccgctgg gcgccccgga
      961 cgccggcgct gccctggccg gcaagctgag gagcggcgac cgcagcatgg tggaggtgct
     1021 ggccgaccac ccgggcgagc tggtgcgcac cgacagcccc aacttcctct gctccgtgct
     1081 gcctacgcac tggcgctgca acaagaccct gcccatcgct ttcaaggtgg tggccctagg
     1141 ggatgttcca gatggcactc tggtcactgt gatggctggc aatgatgaaa actactcggc
     1201 tgagctgaga aatgctaccg cagccatgaa gaaccaggtt gcaagattta atgacctcag
     1261 gtttgtcggt cgaagtggaa gagggaaaag cttcactctg accatcactg tcttcacaaa
     1321 cccaccgcaa gtcgccacct accacagagc catcaaaatc acagtggatg ggccccgaga
     1381 acctcgatgt ttaagctatg aaacggaaat attgacagta gaatatggat tattaccgat
     1441 tggtaaaatt gtagaaaagc gcatcgaatg tactgtttat agcgttgata ataatggaaa
     1501 tatttataca caacctgtag cacaatggca cgatcgcgga aagcaaaagg tgtttgagta
     1561 ttgtttggag gatggttcat tgattcgggc aacaaaagac cataagttta tgactgttga
     1621 tggacaaatg ttgccaatta aggaaatatt tcgtcgtaag ttggatttga tgcgggttga
     1681 taatttgccg aatggaagcg gaggagaaaa tctatacttc caaggtgaga acctgtattt
     1741 tcagggcggg agtggtggga tcgaaatagc cacagaaaaa tatttaggcg aacaaaatgt
     1801 ctatgacatt ggagttgagc gcgaccataa ttttgcactc aaaaatggcg gctattttca
     1861 gggcatcgaa atagccacag aaaaatattt aggcgaacaa aatgtctatg acattggagt
     1921 tgagcgcgac cataattttg cactcaaaaa tggcaacagc gccggcggag tgcaggtgga
     1981 aaccatctcc ccaggagacg ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca
     2041 ctacaccggg atgcttgaag atggaaagaa atttgattcc tcccgggaca gaaacaagcc
     2101 ctttaagttt atgctaggca agcaggaggt gatccgaggc tgggaagaag gggttgccca
     2161 gatgagtgtg ggtcagagag ccaaactgac tatatctcca gattatgcct atggtgccac
     2221 tgggcaccca ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg agcttctaaa
     2281 actggaagaa cagaaactga ttagtgagga agatggagga agtggaggta gtgctctcca
     2341 aaaaaagttg gaagagcttg aactggacga acagtagagg atcccgggtg gcatccctgt
     2401 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     2461 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     2521 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     2581 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     2641 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     2701 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     2761 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     2821 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt aaaataacta
     2881 taccagcagg aggacgtcca gacacagcat aggctacctg gccatgccca accggtggga
     2941 catttgagtt gcttgcttgg cactgtcctc tcatgcgttg ggtccactca gtagatgcct
     3001 gttgaattgg gtacgcggcc agcttggctg tggaatgtgt gtcagttagg gtgtggaaag
     3061 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3121 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3181 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3241 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3301 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3361 tgcaaaaagc tcctcgagga actgaaaaac cagaaagtta attccctata gtgagtcgta
     3421 ttaaattcgt aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3481 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3541 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3601 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3661 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3721 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     3781 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     3841 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     3901 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     3961 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4021 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4081 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4141 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4201 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4261 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4321 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4381 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4441 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4501 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4561 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4621 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4681 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4741 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     4801 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     4861 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4921 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4981 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5041 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5101 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5161 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5221 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5281 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5341 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5401 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5461 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5521 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5581 tgccacctga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     5641 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     5701 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     5761 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     5821 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     5881 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     5941 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     6001 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttccatt cgccattcag
     6061 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     6121 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     6181 acgtt
//
