LOCUS       sequence-481777-        6986 bp    DNA     circular SYN 04-AUG-2025
DEFINITION  Proximity-triggered Protein Trans-Splicing to generate the splice 
            product MLL-AF9, Figure 2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6986)
  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 6986)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 4, 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-NUP98n-CAGEn-NES (Nrdj1)
FEATURES             Location/Qualifiers
     source          1..6986
                     /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             2768..3088
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    3187..3809
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3386..3405)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3833..3853)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3838..4167
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4018..4153
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4205..4224)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4206..4224)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4238..4254)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4238..4254)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4251..4273)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4262..4278
                     /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(4286..4292,4293..4310,4311..4316))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4331..4352
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4469..4486)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4640..5228)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4720..4739)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5399..6190,6191..6259))
                     /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     6022..6041
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6260..6364)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6391..6846
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6478..6497)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6688..6709
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     join(6972..6986,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(6986,1..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 gttgtgatgt ttaacaaatc atttggaaca ccctttgggg gtggcacagg
      901 tggctttggc acaacttcaa catttggaca gaatactggc tttggcacta ctagtggagg
      961 ggcatttgga acatctgcat ttggttctag caacaatact ggaggcctct ttggaaattc
     1021 acagactaaa ccaggaggat tgtttggaac cagttcattt agccagccag ctacctccac
     1081 aagcactggc tttgggtttg gtacgtcaac aggaacagca aataccttgt ttggaactgc
     1141 aagcacaggg accagtctct tctcatccca aaacaatgcc tttgcacaaa ataaaccaac
     1201 tggctttggc aattttggaa ccagtactag cagtggagga ctctttggaa ccacaaatac
     1261 cacctctaat ccttttggca gcacatctgg ctccctcttt gggccaagta gttttacagc
     1321 tgctcctact gggactacta ttaaatttaa ccctccaact ggtacagata ctatggtcaa
     1381 agctggagtt agcactaaca taagtaccaa gcaccagtgt attactgcta tgaaagaata
     1441 tgaaagcaag tcactagagg aacttcgttt agaggattat caggctaaca ggaagggccc
     1501 acagaaccag gtgggagcag gtaccacaac tggcttgttt gggtcttctc cagccacttc
     1561 cagcgcaaca ggactcttca gctcctccac cactaattca ggctttgcat atggtcagaa
     1621 caaaactgcc tttggaacta gtacaactgg atttggaaca aatccaggtg gtctctttgg
     1681 ccaacagaat cagcagacta ccagcctctt cagcaaacca tttggccagg ctacaaccac
     1741 ccagaacact ggcttttcct ttggtaatac cagcaccata ggacagccaa gcaccaacac
     1801 catgggatta tttggagtaa cccaagcctc acagcctgga ggtctttttg ggacagctac
     1861 aaacaccagc actgggacag catttggaac aggaacaggt ctctttgggc agaccaatac
     1921 tggatttggt gctgttggtt cgaccctgtt tggcaataac aagcttacta catttggaag
     1981 cagcacaacc agtgcacctt catttggtac aaccagtggc gggctctttg gttttggcac
     2041 aaataccagt gggaatagta tttttggaag taaaccagca cctgggactc ttggaactgg
     2101 gcttggtgca ggatttggaa cagctcttgg tgctggacag gcatctttgt ttgggaacaa
     2161 ccaacctaag attggagggc ctcttggtac aggagccttt ggggcccctg gatttaatac
     2221 tacgacagcc actttgggct ttggagcccc ccaggcccca gtaagagaaa aacaacccag
     2281 cgaaggcgcc ttctgtctgg tcggctctag cgaaatcatc acacgtaatt atgggaagac
     2341 aaccattaag gaagtagttg agatatttga caatgataaa aatatccaag ttctagcgtt
     2401 taacactcac actgacaaca tcgagtgggc tcccattaaa gctgcacaac ttactagacc
     2461 gaacgctgag ttggttgaac tcgaaattga taccctacat ggtgtgaaaa cgatcaggtg
     2521 tacccccgac cacccagttt acactaaaaa caggggctac gtgcgagcgg atgagttaac
     2581 cgatgatgat gagcttgttg tagccatcga gaacctgtat ggcgatgaag tagatggcta
     2641 ttttcagggc atggaagcgg agacgtatat aggaaagctt aaatcccgag caattgtatc
     2701 aaacgaggac acgtatgata ttcagacgtc aactcataac ttttttgcga atgataacag
     2761 cgccggcgga gtgcaggtgg aaaccatctc cccaggagac gggcgcacct tccccaagcg
     2821 cggccagacc tgcgtggtgc actacaccgg gatgcttgaa gatggaaaga aatttgattc
     2881 ctcccgggac agaaacaagc cctttaagtt tatgctaggc aagcaggagg tgatccgagg
     2941 ctgggaagaa ggggttgccc agatgagtgt gggtcagaga gccaaactga ctatatctcc
     3001 agattatgcc tatggtgcca ctgggcaccc aggcatcatc ccaccacatg ccactctcgt
     3061 cttcgatgtg gagcttctaa aactggaaga acagaaactg attagtgagg aagatggagg
     3121 aagtggaggt agtgctctcc aaaaaaagtt ggaagagctt gaactggacg aacagtagag
     3181 gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     3241 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     3301 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     3361 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     3421 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     3481 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     3541 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     3601 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     3661 cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca taggctacct
     3721 ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt
     3781 gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg
     3841 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3901 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3961 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4021 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4081 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4141 ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt
     4201 aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa
     4261 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4321 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4381 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4441 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4501 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4561 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4621 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4681 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4741 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     4801 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     4861 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     4921 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     4981 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5041 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5101 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5161 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5221 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5281 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5341 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5401 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5461 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5521 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5581 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5641 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5701 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     5761 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     5821 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     5881 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     5941 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6001 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6061 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6121 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6181 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6241 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6301 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6361 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     6421 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     6481 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     6541 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     6601 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     6661 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     6721 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     6781 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     6841 caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
     6901 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
     6961 gccagggttt tcccagtcac gacgtt
//
