LOCUS       sequence-485488-        6465 bp    DNA     circular SYN 08-SEP-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 6465)
  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 6465)
  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: pcDNA3.1-NES-CAGEc-HOXA9c (Nrdj1)
FEATURES             Location/Qualifiers
     source          1..6465
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             988..1011
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1021..1044
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1054..1077
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1114..1389
                     /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             2005..2031
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2056..2073)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2062..2286
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2332..2760
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2419..2438)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2629..2650
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2769..2789)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2774..3103
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2954..3089
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3016..3035
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3170..3964
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3224..3243)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3834..3853
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4138..4259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4175..4194)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4229..4248
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4308..4324)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4308..4324)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4321..4343)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4332..4348
                     /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(4356..4362,4363..4380,4381..4386))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4401..4422
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4539..4556)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4710..5298)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4790..4809)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5469..6260,6261..6329))
                     /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     6092..6111
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6330..6434)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccatggctc tccaaaaaaa gttggaagag cttgaactgg acgaacaggg aggaagtgga
      961 ggtagtctcg agggtaccat gggttcagac tataaagatg atgatgataa aggtggctcc
     1021 gactacaagg atgacgatga caagggaggt agtgactaca aagacgatga cgacaagggc
     1081 ggaggaggaa gtggaggtag tggaagagta gcgatcctct ggcatgagat gtggcatgaa
     1141 ggcctggaag aggcatctcg tttgtacttt ggggaaagga acgtgaaagg catgtttgag
     1201 gtgctggagc ccttgcatgc tatgatggaa cggggccccc agactctgaa ggaaacatcc
     1261 tttaatcagg cctatggtcg agatttaatg gaggcccaag agtggtgcag gaagtacatg
     1321 aaatcaggga atgtcaagga cctcctccaa gcctgggacc tctattatca tgtgttccga
     1381 cgaatctcag gaaatggaaa tggaaacgct gagttggttg aactcgaaat taagacccta
     1441 catggtgtga aaacgatcag gtgtaccccc gaccacccag tttacactaa aaacaggggc
     1501 tacgtgcgag cggatgagtt aaccgatgat gatgagcttg ttgtagccat cgagaacctg
     1561 tatggcgatg aagtagatgg ctattttcag ggcatggaag cgaagacgta tataggaaag
     1621 cttaaatccc gaaaaattgt atcaaacgag gacacgtatg atattcagac gtcaactcat
     1681 aacttttttg cgaatgatat acttgttcat aattctgaaa acaatgctga gaatgagagc
     1741 ggcggagaca agccccccat cgatcccaat aacccagcag ccaactggct tcatgcgcgc
     1801 tccactcgga aaaagcggtg cccctataca aaacaccaga ccctggaact ggagaaagag
     1861 tttctgttca acatgtacct caccagggac cgcaggtacg aggtggctcg actgctcaac
     1921 ctcaccgaga ggcaggtcaa gatctggttc cagaaccgca ggatgaaaat gaagaaaatc
     1981 aacaaagacc gagcaaaaga cgagtacccg tacgacgttc cggactacgc ttcttgagtt
     2041 taaaccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     2101 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     2161 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     2221 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     2281 ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct
     2341 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     2401 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     2461 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     2521 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2581 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2641 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2701 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2761 aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     2821 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     2881 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     2941 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3001 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     3061 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     3121 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3181 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3241 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3301 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     3361 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3421 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3481 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3541 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3601 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     3661 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     3721 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     3781 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     3841 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     3901 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     3961 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4021 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4081 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     4141 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4201 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4261 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     4321 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4381 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4441 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4501 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4561 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4621 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4681 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4741 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4801 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4861 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4921 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4981 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5041 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5101 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5161 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5221 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5281 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5341 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5401 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5461 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5521 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5581 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5641 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5701 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5761 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5821 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5881 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5941 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6001 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6061 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6121 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6181 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6241 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6301 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6361 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6421 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
