LOCUS       sequence-413183-        6252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammailan expression of WDR5, SH2, and HRas separated by P2A..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6252)
  AUTHORS   Sekhon H, Ha JH, Presti MF, Procopio SB, Jarvis AR, Mirsky PO, John 
            AM, Loh SN
  TITLE     Adaptable, turn-on maturation (ATOM) fluorescent biosensors for 
            multiplexed detection in cells.
  JOURNAL   Nat Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w.
  PUBMED    37945909
REFERENCE   2  (bases 1 to 6252)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w."
COMMENT     Sequence Label: pCMV-WDR5-P2A-SH2-P2A-Ras-CAAX
FEATURES             Location/Qualifiers
     source          1..6252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             612..635
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1749..1805
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             2211..2267
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     polyA_signal    3040..3161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3077..3096)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3131..3150
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3168..3623)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3305..3326)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3517..3536
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3650..3754
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3756..4113
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3756..3947
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3779..3799)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3964..4099
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4148..4942
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4148..4942
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4202..4221)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4812..4831
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5130..5149)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5174..5221
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5550..6138
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6039..6058
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccgccaccat gttagaagtg ctgtttcaag gtccaggcat ggtgcccatc ggagccgtgc
      661 acggcggcca tcccggcgta gtgcatccgc cacagcaacc actgcccacg gcgcccagcg
      721 gcccaaactc gctgcagccg aactcggtgg gccagccggg ggccaccacc tcctcgaaca
      781 gcagcgcctc caacaagagc tcgctatccg tcaagcccaa ctacacgctc aagttcacgc
      841 tggccgggca caccaaggcg gtgtcggcgg tcaagttcag tccgaatggc gagtggctgg
      901 ccagctcctc cgctgataaa ctaatcaaaa tctggggagc atacgatggc aagttcgaga
      961 agaccatttc gggccacaag ctgggcatca gcgatgtggc ctggagctca gactcgcgac
     1021 tcctcgtgag cggcagtgat gacaagacgc tcaaggtctg ggagctgagc accgggaaga
     1081 gcttgaaaac tctgaagggc cacagcaact atgtgttctg ctgcaacttt aatccgcagt
     1141 ccaatctgat cgtctccggc agcttcgacg agagcgttcg catatgggat gtgcgcaccg
     1201 gcaagtgtct gaagactcta cccgcccatt ccgatcccgt ttcggcggta catttcaatc
     1261 gcgacggatc gctgatcgtg agcagcagct acgacggcct ctgtcgcata tgggacacgg
     1321 ccagtggaca gtgcttgaaa accctgatcg acgacgacaa tccgcccgtc agctttgtaa
     1381 agttctcgcc caatggcaag tacattttgg ccgccacgct ggataatacg ctcaagttgt
     1441 gggactactc gaagggcaag tgcctgaaga cgtatacggg tcacaagaat gagaagtact
     1501 gcatattcgc caacttctcg gtgacgggag gaaagtggat cgtgagtggc agcgaggaca
     1561 acatggtcta catttggaat ctgcagagca aggaggtggt gcaaaagctg cagggacaca
     1621 ccgataccgt tctgtgcacc gcctgccatc ccacggagaa catcattgct tccgcggcgc
     1681 tcgagaacga caagaccatc aagctgtgga agtcggatac aggactcaga tctcgagctg
     1741 gtagcggtgc tactaacttc agcctgctga agcaggctgg agacgtggag gagaaccctg
     1801 gacctggtag cggtccgaat tcggccacca tgccaagcaa ctacatcacg ccagtcaaca
     1861 gtctggagaa acactcctgg taccatgggc ctgtgtcccg caatgccgct gagtatctgc
     1921 tgagcagcgg gatcaatggc agcttcttgg tgcgtgagag tgagagcagt cctggccaga
     1981 ggtccatctc gctgagatac gaagggaggg tgtaccatta caggatcaac actgcttctg
     2041 atggcaagct ctacgtctcc tccgagagcc gcttcaacac cctggccgag ttggttcatc
     2101 atcattcaac ggtggccgac gggctcatca ccacgctcca ttatccagcc ccaaagcgca
     2161 acaagcccac tgtctatggt gtgtccccca actacaagct tggtagcggt gctactaact
     2221 tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctggt agcggtggat
     2281 ccatgacaga atacaagtta gtagtcgtcg gtgctgttgg tgttggtaaa tccgctttga
     2341 caatacaatt aatacaaaat cacttcgtag atgaatacga tccaactatc gaagattcat
     2401 acagaaagca agttgttatt gatggtgaaa cttgtttgtt ggatatcttg gatacagctg
     2461 gtcaagaaga atattctgca atgagagatc aatacatgag aactggtgaa ggtttcttgt
     2521 gtgtttttgc tattaataac acaaaatcat ttgaagatat tcatcaatat agagaacaaa
     2581 ttaaaagagt taaagattct gatgatgttc caatggtttt ggttggtaat aagtgtgatt
     2641 tggctgcaag aactgttgaa tctagacaag ctcaagattt ggcaagatct tatggtattc
     2701 catacattga aacttctgct aaaacaagac aaggtgttga agatgcattc tacacattgg
     2761 ttagagaaat cagacaacat ggtggaacta gtggttctgg cggtgctggt gcatcaggcg
     2821 gtagtggtac tggtggaggc gctagcaaat tgagaaagct gaaccctcct gatgagagtg
     2881 gccccggctg catgagctgc aagtgtgtgc tctcctaagc ggccgcgact ctagatcata
     2941 atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc
     3001 ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat
     3061 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     3121 cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt
     3181 taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata
     3241 ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt
     3301 tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg
     3361 aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt
     3421 ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc
     3481 ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg
     3541 cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct
     3601 taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3661 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3721 ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg
     3781 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3841 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     3901 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     3961 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     4021 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     4081 ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc
     4141 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     4201 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     4261 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     4321 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     4381 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     4441 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     4501 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     4561 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     4621 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     4681 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     4741 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     4801 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     4861 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     4921 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     4981 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     5041 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     5101 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     5161 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     5221 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     5281 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     5341 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     5401 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5461 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5521 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5581 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5641 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     5701 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5761 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5821 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     5881 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     5941 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6001 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6061 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6121 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6181 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6241 accgccatgc at
//
