LOCUS       sequence-403675-        7861 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of RUNX4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7861)
  AUTHORS   Oei V, Chuang LSH, Matsuo J, Srivastava S, Teh M, Ito Y
  TITLE     RUNX3 inactivates oncogenic MYC through disruption of MYC/MAX 
            complex and subsequent recruitment of GSK3beta-FBXW7 cascade.
  JOURNAL   Commun Biol. 2023 Jul 3;6(1):689. doi: 10.1038/s42003-023-05037-0.
  PUBMED    37400551
REFERENCE   2  (bases 1 to 7861)
  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: "Commun 
            Biol."; date: "2023-07-3"; volume: "6(1):689. doi"; pages: " 
            10.1038/s42003-023-05037-0"
COMMENT     Sequence Label: pRetroX-Tight-puro-3xFLAG-RUNX3 R122C
FEATURES             Location/Qualifiers
     source          1..7861
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     327..350
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     regulatory      604..613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1659..1724)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        complement(1747..2061)
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     primer_bind     complement(1750..1774)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    1821..1839
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1857..1875
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1892..1910
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1928..1946
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1964..1982
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1999..2017
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2035..2053
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             complement(2097..2513)
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     complement(2113..2129)
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(2145..2167)
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     misc_feature    2714..2913
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     LTR             2976..3151
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     promoter        complement(3152..3355)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     enhancer        3356..3735
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3835..3853)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3921..4025
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4026..4094,4095..4886)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4244..4263)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5057..5645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5546..5565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5799..5816
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(5897..5916)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        complement(5938..6267)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5952..6087
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6252..6272
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     6408..6430
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     LTR             6533..6958
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     CDS             complement(7139..7738)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(7222..7242)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7719..7738
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(join(7759..7861,1..385))
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7807..7826)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
ORIGIN
        1 cccctgagcc nncccccgga cccacccctt cccagcctct gagcccagaa agcgaaggag
       61 caaagctgct attggccgct gccccaaagg cctacccgct tccattgctc agcggtgctg
      121 tccatctgca cgagactagt gagacgtgct acttccattt gtcacgtcct gcacgacgcg
      181 agctgcgggg cgggggggaa cttcctgact aggggaggag tagaaggtgg cgcgaagggg
      241 ccaccaaaga acggagccgg ttggcgccta ccggtggatg tggaatgtgt gcgaggccag
      301 aggccacttg tgtagcgcca agtgcccagc ggggctgcta aagcgcatgc tccagactgc
      361 cttgggaaaa gcgcctcccc tacccggtag aattctcagt agggccgcca cacggcctca
      421 tccatgcggc ctggcgtgct cagggccgtg ggtgagttgc tgtggctgcc gtcggcctcc
      481 acgccatcac tctggccgcc caggctgggg ttcatgaggt tgccggcggc gacagaggca
      541 gcgctgctgg tgcaagaggc cagcatgcgg gtaggtgagc ggtcgccccc actgctgctg
      601 ccggccacca tggagaactg gtaggagcca gaggatgtcc cgtagtagag gtggtagggg
      661 gacgggttgg cctggaaggg cccgctctgg ttctgcgggg cccccgggta gggtggcggg
      721 aggtaggtat ggtggaagcg gctggtggcc ggcatgcccg ccacgctgag gctgctgatg
      781 ctcgtgcccg agggcgtggc gctgtagggg aaggcagctg acatggcccc gggataatgc
      841 atcctggggt ctgggaagcg gctctccgtg agggttggca gcgtggggaa ggagcggtca
      901 aactggcggg ggtcggagaa cggatttaat tccgaggtgc cttggattgg ggtctggggc
      961 tggctgctga agtggcttgt ggtgctgagt gagcctcggg ggctgggtgt gctcggtgtc
     1021 acccgcatgc gcagccgttc caggtcccca aagcggtcag ggaacggctt ggtctggtcc
     1081 tccagcttct gccggtgccg tctgggctcc cggggtccgt ccacggtcac cttgatggct
     1141 cggtggtagg tcgccacttg ggtggggttg gtgaacacag tgatggtcag ggtgaaactc
     1201 ttccctcgcc cactgcggcc cacgaagcga aggtcgttga acctggccac ctggttcttc
     1261 atgacggccg aggcattgca cagctcagcg gagtagttct cgtcattgcc tgccatcaca
     1321 gtcaccaccg taccatccgg cacgtccccc aatgccacca ccttgaaggc gacgggcagc
     1381 gtcttgttgc agcgccagtg cgagggcagc acggagcaga ggaagttggg gctgtcggtg
     1441 cgcacgagct cgcctgcgtg gtccgccagc acgtccacca tcgagcgcac ctcgggccgg
     1501 gcgcgccctc cgggccccac ggccgcctgc gcgctcagcg cgccgctgtt ctcgcccatc
     1561 ttgccgccgc cgccgccgca ggggaaggcc ggggagggag gtgtgaagcg gcggctggtg
     1621 cttgggtcta cgggaatacg catcgcggcc gcggatccct tgtcatcgtc atccttgtaa
     1681 tcgatgtcat gatctttata atcaccgtca tggtctttgt agtccatgtc gacagatcct
     1741 tctccaggcg atctgacggt tcactaaacg agctctgctt atataggcct cccaccgtac
     1801 acgcctacct cgacatacgt tctctatcac tgatagggag taaactcgac atacgttctc
     1861 tatcactgat agggataaac tcgacatacg ttctctatca ctgataggga gtaaactcga
     1921 catacgttct ctatcactga tagggagtaa actcgacata cgttctctat cactgatagg
     1981 gagtaaactc gacatcgttc tctatcactg atagggagta aactcgacat acgttctcta
     2041 tcactgatag ggagtaaact cgagtgaaga cgaaagggcc tcagatcttc aattccggcg
     2101 cctagagaag gagtgagggc tggataaagg gaggatcgag gcggggtcga acgaggaggt
     2161 tcaaggggga gagacggggc ggatggagga agaggaggcg gaggcttagg gtgtacaaag
     2221 ggcttgaccc agggaggggg gtcaaaagcc aaggcttccc aggtcacgat gtaggggacc
     2281 tggtctgggt gtccatgcgg gccaggtgaa aagaccttga tcttaacctg ggtgatgagg
     2341 tctcggttaa aggtgccgtc tcgcggccat ccgacgttaa aggttggcca ttctgcagag
     2401 cagaaggtaa cccaacgtct cttcttgaca tctaccgact ggttgtgagc gatccgctcg
     2461 acatctttcc agtgacctaa ggtcaaactt aagggagtgg taacagtctg gccctaattt
     2521 tcagacaaat acagaaacac agtcagacag agacaacaca gaacgatgct gcagcgctgc
     2581 agcagacaag acgcgcggct tcggttccaa accgaaagca aaaattcaga cggaggcggg
     2641 aactgtttta ggttctcgtc tcctaccaga accacatatc ctgacggggt cggattccac
     2701 atcgactccc ttcctcaggt cgggccacaa aaacggcccc caaagtccct gggacgtctc
     2761 ccagggttgc ggccgggtgt tcagaactcg tcagttccac cacgggtccg ccagatacag
     2821 agctagttag ctaactagta cagacgcagg cgcataacat caaacataga cactagacaa
     2881 tcggacagac acagataagt tgctggccag cttacctccc ggtggtgggt cggtggtccc
     2941 tgggcagggg tctccaaatc ccggacgagc ccccaaatga aagacccccg tcgtgggtag
     3001 tcaatcactc agaggagacc ctcccaagga acagcgagac cacgattcgg atgcaaacag
     3061 caagaggctt tattgggaat acgggtaccc gggcgacgca gtctatcgga agactggcgc
     3121 gccgagtgag gggttgtggg ctcttttatt gagctctgct tatatagacc tcccaccgta
     3181 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     3241 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     3301 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     3361 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     3421 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     3481 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     3541 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     3601 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     3661 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     3721 tcaataatca atgtcaacat ggcggtaatg ttggacatga gccaatataa atgtacatat
     3781 tatgatatgg atacaacgta tgcaatggcc aagctaattc ttgaagacga aagggcctcg
     3841 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3901 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3961 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4021 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4081 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4141 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4201 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4261 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4321 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4381 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4441 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4501 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4561 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4621 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4681 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4741 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4801 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4861 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4921 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4981 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5041 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5101 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5161 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     5221 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     5281 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5341 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5401 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5461 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5521 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5581 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5641 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5701 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     5761 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     5821 cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     5881 tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag taagcttttt
     5941 gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     6001 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     6061 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga
     6121 ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca
     6181 cctggttgct gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg
     6241 ggactttcca caccctaact gacacacatt ccacagccgg atctacactc cgctatcgct
     6301 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     6361 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     6421 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagccagctt acctcccggt
     6481 ggtgggtcgg tggtccctgg gcaggggtct cccgatcccg gacgagcccc caaatgaaag
     6541 acccccgctg acgggtagtc aatcactcag aggagaccct cccaaggaac agcgagacca
     6601 caagtcggat gcaactgcaa gagggtttat tggatacacg ggtacccggg cgactcagtc
     6661 aatcggagga ctggcgcccc gagtgagggg ttgtgggctc ttttattgag ctcggggagc
     6721 agaagcgcgc gaacagaagc gagaagcgaa ctgattggtt agttcaaata aggcacaggg
     6781 tcatttcagg tccttggggc accctggaaa catctgatgg ttctctaggg tcgaccctgt
     6841 tccatctgtt cctgaccttg atctgaactt ctctattctc agttatgtat ttttccatgc
     6901 cttgcaaaat ggcgttactt aagctagctt gccaaaccta caggtggggt ctttcattcc
     6961 cccctttttc tggagactaa ataaaatctt ttattttatc tatggctcgt actctatagg
     7021 cttcagctgg tgatattgtt gagtcaaaac tagagcctgg accactgata tctcgagacg
     7081 ccgatgcatg gggtcgtgcg ctcctttcgg tcgggcgctg cgggtcgtgg ggcgggcgtc
     7141 aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg
     7201 cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca
     7261 ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc
     7321 ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct
     7381 ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac
     7441 cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct
     7501 ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt
     7561 cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt
     7621 ccggatcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta
     7681 cggccctggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg
     7741 taagcttggg ctgcaggtcg aaaggcccgg agatgaggaa gaggagaaca gcgcggcaga
     7801 cgtgcgcttt tgaagcgtgc agaatgccgg gcctccggag gaccttcggg cgcccgcccc
     7861 g
//
