LOCUS       sequence-149934-        6876 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To overexpress MKRN1 S109A  in Mammalian Cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6876)
  AUTHORS   Lee MS, Jeong MH, Lee HW, Han HJ, Ko A, Hewitt SM, Kim JH, Chun KH, 
            Chung JY, Lee C, Cho H, Song J
  TITLE     PI3K/AKT activation induces PTEN ubiquitination and destabilization 
            accelerating tumourigenesis.
  JOURNAL   Nat Commun. 2015 Jul 17;6:7769. doi: 10.1038/ncomms8769.
  PUBMED    26183061
REFERENCE   2  (bases 1 to 6876)
  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 
            Commun."; date: "2015-07-17"; volume: "6:7769. doi"; pages: " 
            10.1038/ncomms8769"
COMMENT     Sequence Label: pcDNA3-FLAG MKRN1 S109A
FEATURES             Location/Qualifiers
     source          1..6876
                     /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"
     regulatory      898..907
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             907..930
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2428..2446)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2429..2446)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2466..2483)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2472..2696
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2742..3170
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2829..2848)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3039..3060
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3179..3199)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3184..3514
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3365..3500
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3427..3446
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3581..4375
                     /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(3635..3654)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4245..4264
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4549..4670
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4586..4605)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4640..4659
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4719..4735)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4719..4735)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4732..4754)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4743..4759
                     /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(4767..4773,4774..4791,4792..4797))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4812..4833
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4950..4967)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5121..5709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5201..5220)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5880..6671,6672..6740))
                     /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     6503..6522
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6741..6845)
                     /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 gcttgccgcc
      901 accatggatt acaaggatga cgacgataag agcgcgggcg gatccgaatt catggcggag
      961 gctgcaactc ccggaacaac agccacaaca tcaggagcag gagcggcagc ggcgacggcg
     1021 gcagcagcct cccccacccc gatccccaca gtcaccgccc cgtccctggg ggctggcgga
     1081 gggggcggcg gcagcgacgg cagcggcggc ggctggacta aacaggtcac ctgcaggtat
     1141 tttatgcatg gggtttgtaa ggaaggagac aactgtcgct actcgcatga cctctctgac
     1201 agtccgtata gtgtagtgtg caagtatttt cagcgagggt actgtattta tggagaccgc
     1261 tgcagatatg aacatgccaa accattgaaa caggaagaag caactgctac agagctaact
     1321 acaaagtcat cccttgctgc ttcctcaagt ctctcatcga tagttggacc acttgttgaa
     1381 atgaatacag gcgaagctga gtcaagaaat tcaaactttg caactgtagg agcaggttca
     1441 gaggactggg tgaatgctat tgagtttgtt cctgggcaac cctactgtgg ccgtactgcg
     1501 ccttcctgca ctgaagcacc cctgcagggc tcagtgacca aggaagaatc agagaaagag
     1561 caaaccgccg tggagacaaa gaagcagctg tgcccctatg ctgcagtggg agagtgccga
     1621 tacggggaga actgtgtgta tctccacgga gattcttgtg acatgtgtgg gctgcaggtc
     1681 ctgcatccaa tggatgctgc ccagagatcg cagcatatca aatcgtgcat tgaggcccat
     1741 gagaaggaca tggagctctc atttgccgtg cagcgcagca aggacatggt gtgtgggatc
     1801 tgcatggagg tggtctatga gaaagccaac cccagtgagc gccgcttcgg gatcctctcc
     1861 aactgcaacc acacctactg tctcaagtgc attcgcaagt ggaggagtgc taagcaattt
     1921 gagagcaaga tcataaagtc ctgcccagaa tgccggatca catctaactt tgtcattcca
     1981 agtgagtact gggtggagga gaaagaagag aagcagaaac tcattctgaa atacaaggag
     2041 gcaatgagca acaaggcgtg caggtatttt gatgaaggac gtgggagctg cccatttgga
     2101 gggaactgtt tttacaagca tgcgtaccct gatggccgta gagaggagcc acagagacag
     2161 aaagtgggaa catcaagcag ataccgggcc caacgaagga accacttctg ggaactcatt
     2221 gaggaaagag agaacagcaa cccctttgac aacgatgaag aagaggttgt cacctttgag
     2281 ctgggcgaga tgttgcttat gcttttggct gcaggtgggg acgacgaact aacagactct
     2341 gaagatgagt gggacttgtt tcatgatgag ctggaagatt tttatgactt ggatctatag
     2401 ctcgagcatg catctagagg gccctattct atagtgtcac ctaaatgcta gagctcgctg
     2461 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     2521 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     2581 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     2641 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     2701 tgaggcggaa agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     2761 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     2821 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     2881 tcaagctcta aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga
     2941 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3001 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3061 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tggggatttc
     3121 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     3181 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc caggcaggca gaagtatgca
     3241 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3301 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3361 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3421 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3481 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3541 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3601 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3661 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3721 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3781 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3841 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3901 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3961 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4021 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4081 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4141 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4201 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4261 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4321 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4381 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4441 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4501 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4561 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4621 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     4681 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     4741 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4801 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4861 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4921 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4981 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5041 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5101 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5161 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5221 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5281 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     5341 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5401 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5461 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5521 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     5581 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5641 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5701 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5761 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5821 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5881 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5941 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6001 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6061 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6121 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6181 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6241 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6301 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6361 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6421 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6481 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6541 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6601 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6661 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6721 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6781 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6841 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
