LOCUS       sequence-403674-        6612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of RUNX3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6612)
  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 6612)
  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: pcFLAG RUNX3 (R122C)
FEATURES             Location/Qualifiers
     source          1..6612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(151..169)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(152..169)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(189..206)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    195..419
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      465..893
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(552..571)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     762..783
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(902..922)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        946..1156
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        968..1164
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1015..1150
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1077..1096
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             1231..2025
                     /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(1285..1304)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1895..1914
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2199..2320
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2236..2255)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2290..2309
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2369..2385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2369..2385)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2382..2404)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2393..2409
                     /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(2417..2423,2424..2441,2442..2447))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2462..2483
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2600..2617)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2771..3359)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2851..2870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3530..4321,4322..4390))
                     /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     4153..4172
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4391..4495)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4570..4589)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4761..5140
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5141..5344
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5295..5315
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5389..5408
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5389..5407
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5448..5471
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      6511..6520
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 accccagcct gggcggccag agtgatggcg tggaggccga cggcagccac agcaactcac
       61 ccacggccct gagcacgcca ggccgcatgg atgaggccgt gtggcggccc tactgatcta
      121 gagcggccgc tcgagtctag agggccctat tctatagtgt cacctaaatg ctagagctcg
      181 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
      241 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
      301 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
      361 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
      421 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
      481 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
      541 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
      601 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
      661 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
      721 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
      781 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
      841 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
      901 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
      961 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     1021 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     1081 tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt
     1141 tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct
     1201 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     1261 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     1321 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     1381 accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg
     1441 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     1501 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     1561 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     1621 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     1681 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     1741 ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     1801 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     1861 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     1921 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     1981 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     2041 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     2101 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     2161 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     2221 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2281 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     2341 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2401 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2461 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2521 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2581 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2641 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2701 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2761 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2821 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2881 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2941 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3001 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3061 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3121 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3181 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3241 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3301 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3361 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3421 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3481 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3541 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3601 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3661 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3721 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3781 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3841 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3901 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3961 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4021 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4081 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4141 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4201 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4261 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4321 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4381 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4441 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4501 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     4561 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     4621 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     4681 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     4741 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac
     4801 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     4861 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     4921 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     4981 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     5041 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     5101 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     5161 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     5221 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     5281 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     5341 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
     5401 ctatagggag acccaagctt ggtaccgagc tcggatccac cgccatggac tacaaggacg
     5461 atgatgacaa gggcgaattc gggcgtattc ccgtagaccc aagcaccagc cgccgcttca
     5521 cacctccctc cccggccttc ccctgcggcg gcggcggcgg caagatgggc gagaacagcg
     5581 gcgcgctgag cgcgcaggcg gccgtggggc ccggagggcg cgcccggccc gaggtgcgct
     5641 cgatggtgga cgtgctggcg gaccacgcag gcgagctcgt gcgcaccgac agccccaact
     5701 tcctctgctc cgtgctgccc tcgcactggc gctgcaacaa gacgctgccc gtcgccttca
     5761 aggtggtggc attgggggac gtgccggatg gtacggtggt gactgtgatg gcaggcaatg
     5821 acgagaacta ctccgctgag ctgtgcaatg cctcggccgt catgaagaac caggtggcca
     5881 ggttcaacga ccttcgcttc gtgggccgca gtgggcgagg gaagagtttc accctgacca
     5941 tcactgtgtt caccaacccc acccaagtgg cgacctacca ccgagccatc aaggtgaccg
     6001 tggacggacc ccgggagccc agacggcacc ggcagaagct ggaggaccag accaagccgt
     6061 tccctgaccg ctttggggac ctggaacggc tgcgcatgcg ggtgacaccg agcacaccca
     6121 gcccccgagg ctcactcagc accacaagcc acttcagcag ccagccccag accccaatcc
     6181 aaggcacctc ggaactgaac ccattctccg acccccgcca gtttgaccgc tccttcccca
     6241 cgctgccaac cctcacggag agccgcttcc cagaccccag gatgcattat cccggggcca
     6301 tgtcagctgc cttcccctac agcgccacgc cctcgggcac gagcatcagc agcctcagcg
     6361 tggcgggcat gccggccacc agccgcttcc accataccta cctcccgcca ccctacccgg
     6421 gggccccgca gaaccagagc gggcccttcc aggccaaccc gtccccctac cacctctact
     6481 acgggacatc ctctggctcc taccagttct ccatggtggc cggcagcagc agtgggggcg
     6541 accgctcacc tacccgcatg ctggcctctt gcaccagcag cgctgcctct gtcgccgccg
     6601 gcaacctcat ga
//
