LOCUS       sequence-149958-        6873 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To overexpress MKRN1 H307E in Mammalian Cell.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6873)
  AUTHORS   Lee EW, Lee MS, Camus S, Ghim J, Yang MR, Oh W, Ha NC, Lane DP, Song
            J
  TITLE     Differential regulation of p53 and p21 by MKRN1 E3 ligase controls 
            cell cycle arrest and apoptosis.
  JOURNAL   EMBO J. 2009 Jul 22;28(14):2100-13. doi: 10.1038/emboj.2009.164. 
            Epub 2009 Jun 18.
  PUBMED    19536131
REFERENCE   2  (bases 1 to 6873)
  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"; doi: 
            "10.1038/emboj.2009.164"; journalName: "EMBO J"; date: "2009-07-22- 
            22"; volume: "28"; issue: "14"; pages: "2100-13"
COMMENT     Sequence Label: pcDNA3-HA-MKRN1 H307E
FEATURES             Location/Qualifiers
     source          1..6873
                     /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             910..936
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(2425..2443)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2426..2443)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2463..2480)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2469..2693
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2739..3167
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2826..2845)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3036..3057
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3176..3196)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3181..3511
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3362..3497
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3424..3443
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3578..4372
                     /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(3632..3651)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4242..4261
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4546..4667
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4583..4602)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4637..4656
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4716..4732)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4716..4732)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4729..4751)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4740..4756
                     /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(4764..4770,4771..4788,4789..4794))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4809..4830
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4947..4964)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5118..5706)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5198..5217)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5877..6668,6669..6737))
                     /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     6500..6519
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6738..6842)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 atggcctcct acccttatga tgtgccagat tatgcctctc ccgaattcat ggcggaggct
      961 gcaactcccg gaacaacagc cacaacatca ggagcaggag cggcagcggc gacggcggca
     1021 gcagcctccc ccaccccgat ccccacagtc accgccccgt ccctgggggc gggcggaggg
     1081 ggcggcggca gcgacggcag cggcggcggc tggactaaac aggtcacctg caggtatttt
     1141 atgcatgggg tttgtaagga aggagacaac tgtcgctact cgcatgacct ctctgacagt
     1201 ccgtatagtg tagtgtgcaa gtattttcag cgagggtact gtatttatgg agaccgctgc
     1261 agatatgaac atagcaaacc attgaaacag gaagaagcaa ctgctacaga gctaactaca
     1321 aagtcatccc ttgctgcttc ctcaagtctc tcatcgatag ttggaccact tgttgaaatg
     1381 aatacaggcg aagctgagtc aagaaattca aactttgcaa ctgtaggagc aggttcagag
     1441 gactgggtga atgctattga gtttgttcct gggcaaccct actgtggccg tactgcgcct
     1501 tcctgcactg aagcacccct gcagggctca gtgaccaagg aagaatcaga gaaagagcaa
     1561 accgccgtgg agacaaagaa gcagctgtgc ccctatgctg cagtgggaga gtgccgatac
     1621 ggggagaact gtgtgtatct ccacggagat tcttgtgaca tgtgtgggct gcaggtcctg
     1681 catccaatgg atgctgccca gagatcgcag catatcaaat cgtgcattga ggcccatgag
     1741 aaggacatgg agctctcatt tgccgtgcag cgcagcaagg acatggtgtg tgggatctgc
     1801 atggaggtgg tctatgagaa agccaacccc agtgagcgcc gcttcgggat cctctccaac
     1861 tgcaacgaaa cctactgtct caagtgcatt cgcaagtgga ggagtgctaa gcaatttgag
     1921 agcaagatca taaagtcctg cccagaatgc cggatcacat ctaactttgt cattccaagt
     1981 gagtactggg tggaggagaa agaagagaag cagaaactca ttctgaaata caaggaggca
     2041 atgagcaaca aggcgtgcag gtattttgat gaaggacgtg ggagctgccc atttggaggg
     2101 aactgttttt acaagcatgc gtaccctgat ggccgtagag aggagccaca gagacagaaa
     2161 gtgggaacat caagcagata ccgggcccaa cgaaggaacc acttctggga actcattgag
     2221 gaaagagaga acagcaaccc ctttgacaac gatgaagaag aggttgtcac ctttgagctg
     2281 ggcgagatgt tgcttatgct tttggctgca ggtggggacg acgaactaac agactctgaa
     2341 gatgagtggg acttgtttca tgatgagctg gaagattttt atgacttgga tctatagctc
     2401 gagcatgcat ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     2461 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     2521 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     2581 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     2641 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     2701 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     2761 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2821 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2881 agctctaaat cggggcatcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2941 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3001 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3061 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgg ggatttcggc
     3121 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     3181 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag gcaggcagaa gtatgcaaag
     3241 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     3301 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3361 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     3421 ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg
     3481 aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt
     3541 cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3601 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3661 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3721 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc
     3781 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3841 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3901 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3961 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4021 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4081 cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca
     4141 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4201 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4261 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4321 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4381 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4441 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4501 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     4561 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4621 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     4681 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4741 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4801 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4861 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4921 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4981 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5041 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5101 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5161 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5221 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5281 tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg
     5341 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5401 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5461 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5521 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5581 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5641 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5701 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5761 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5821 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5881 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5941 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     6001 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     6061 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     6121 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     6181 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6241 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6301 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6361 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6421 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6481 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6541 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6601 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6661 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6721 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6781 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6841 cgcacatttc cccgaaaagt gccacctgac gtc
//
