LOCUS       sequence-149955-        6267 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To overexpress -MKRN1 1-280 in Mammalian Cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6267)
  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 6267)
  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 1-280
FEATURES             Location/Qualifiers
     source          1..6267
                     /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(1819..1837)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1820..1837)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1857..1874)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1863..2087
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2133..2561
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2220..2239)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2430..2451
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2570..2590)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2575..2905
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2756..2891
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2818..2837
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2972..3766
                     /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(3026..3045)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3636..3655
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3940..4061
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3977..3996)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4031..4050
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4110..4126)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4110..4126)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4123..4145)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4134..4150
                     /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(4158..4164,4165..4182,4183..4188))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4203..4224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4341..4358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4512..5100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4592..4611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5271..6062,6063..6131))
                     /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     5894..5913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6132..6236)
                     /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 acatggtgta actcgagcat
     1801 gcatctagag ggccctattc tatagtgtca cctaaatgct agagctcgct gatcagcctc
     1861 gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac
     1921 cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg
     1981 tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga
     2041 ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga
     2101 aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc
     2161 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc
     2221 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     2281 aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     2341 acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     2401 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     2461 caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt cggcctattg
     2521 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt
     2581 cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc aaagcatgca
     2641 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2701 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2761 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2821 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     2881 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     2941 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3001 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3061 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3121 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     3181 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3241 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3301 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3361 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3421 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3481 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3541 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     3601 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     3661 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     3721 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     3781 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     3841 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     3901 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     3961 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4021 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4081 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4141 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4201 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4261 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4321 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4381 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4441 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4501 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4561 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4621 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4681 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     4741 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4801 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4861 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4921 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     4981 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5041 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5101 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5161 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5221 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5281 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5341 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5401 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5461 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5521 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5581 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5641 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5701 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5761 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5821 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5881 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5941 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6001 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6061 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6121 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6181 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6241 tttccccgaa aagtgccacc tgacgtc
//
