LOCUS       sequence-485482-        6098 bp    DNA     circular SYN 08-SEP-2025
DEFINITION  Proximity-triggered Protein Trans-Splicing to generate the splice 
            product AML1-ETO, Figure 2 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 6098)
  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 6098)
  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-AMLn-CAGEn-NES (Nrdj1)
FEATURES             Location/Qualifiers
     source          1..6098
                     /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             1880..2200
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    2299..2921
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2498..2517)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2945..2965)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2950..3279
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3130..3265
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3317..3336)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3318..3336)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3350..3366)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3350..3366)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3363..3385)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3374..3390
                     /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(3398..3404,3405..3422,3423..3428))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3443..3464
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3581..3598)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3752..4340)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3832..3851)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4511..5302,5303..5371))
                     /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     5134..5153
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5372..5476)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5503..5958
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5590..5609)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5800..5821
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(6084..6098,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6098,1..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 acctcgaaat cgtactgaga agcactgtct ggtcggctct agcgaaatca tcacacgtaa
     1441 ttatgggaag acaaccatta aggaagtagt tgagatattt gacaatgata aaaatatcca
     1501 agttctagcg tttaacactc acactgacaa catcgagtgg gctcccatta aagctgcaca
     1561 acttactaga ccgaacgctg agttggttga actcgaaatt gataccctac atggtgtgaa
     1621 aacgatcagg tgtacccccg accacccagt ttacactaaa aacaggggct acgtgcgagc
     1681 ggatgagtta accgatgatg atgagcttgt tgtagccatc gagaacctgt atggcgatga
     1741 agtagatggc tattttcagg gcatggaagc ggagacgtat ataggaaagc ttaaatcccg
     1801 agcaattgta tcaaacgagg acacgtatga tattcagacg tcaactcata acttttttgc
     1861 gaatgataac agcgccggcg gagtgcaggt ggaaaccatc tccccaggag acgggcgcac
     1921 cttccccaag cgcggccaga cctgcgtggt gcactacacc gggatgcttg aagatggaaa
     1981 gaaatttgat tcctcccggg acagaaacaa gccctttaag tttatgctag gcaagcagga
     2041 ggtgatccga ggctgggaag aaggggttgc ccagatgagt gtgggtcaga gagccaaact
     2101 gactatatct ccagattatg cctatggtgc cactgggcac ccaggcatca tcccaccaca
     2161 tgccactctc gtcttcgatg tggagcttct aaaactggaa gaacagaaac tgattagtga
     2221 ggaagatgga ggaagtggag gtagtgctct ccaaaaaaag ttggaagagc ttgaactgga
     2281 cgaacagtag aggatcccgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc
     2341 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt
     2401 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag
     2461 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt
     2521 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg
     2581 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg
     2641 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct
     2701 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct
     2761 cccttccctg tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag
     2821 cataggctac ctggccatgc ccaaccggtg ggacatttga gttgcttgct tggcactgtc
     2881 ctctcatgcg ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcttgg
     2941 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3001 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3061 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3121 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3181 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     3241 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcctcga ggaactgaaa
     3301 aaccagaaag ttaattccct atagtgagtc gtattaaatt cgtaatcatg tcatagctgt
     3361 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     3421 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     3481 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3541 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3601 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3661 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3721 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3781 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3841 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3901 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3961 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4021 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4081 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4141 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4201 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4261 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4321 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4381 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4441 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4501 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4561 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4621 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4681 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4741 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4801 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4861 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4921 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4981 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5041 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5101 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5161 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5221 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5281 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5341 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5401 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5461 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg
     5521 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     5581 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5641 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5701 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5761 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5821 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5881 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5941 tattaacgct tacaatttcc attcgccatt caggctgcgc aactgttggg aagggcgatc
     6001 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
     6061 aagttgggta acgccagggt tttcccagtc acgacgtt
//
