LOCUS       sequence-232079-        8252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CIP2A L533E V5 overexpression, CIP2A dimer defective mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8252)
  AUTHORS   Wang J, Okkeri J, Pavic K, Wang Z, Kauko O, Halonen T, Sarek G, 
            Ojala PM, Rao Z, Xu W, Westermarck J
  TITLE     Oncoprotein CIP2A is stabilized via interaction with tumor 
            suppressor PP2A/B56.
  JOURNAL   EMBO Rep. 2017 Mar;18(3):437-450. doi: 10.15252/embr.201642788. Epub
            2017 Feb 7.
  PUBMED    28174209
REFERENCE   2  (bases 1 to 8252)
  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: 
            "10.15252/embr.201642788"; journalName: "EMBO Rep"; date: "2017-03";
            volume: "18"; issue: "3"; pages: "437-450"
COMMENT     Sequence Label: pcDNA3.1/CIP2A(1-905) L533E V5 His
FEATURES             Location/Qualifiers
     source          1..8252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3516..3557
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3567..3584
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3607..3624)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3613..3837
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3883..4311
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3970..3989)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4180..4201
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4320..4340)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4325..4654
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4505..4640
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4567..4586
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4721..5515
                     /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(4775..4794)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5385..5404
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5691..5812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5728..5747)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5782..5801
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5861..5877)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5861..5877)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5874..5896)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5885..5901
                     /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(5909..5915,5916..5933,5934..5939))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5954..5975
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6092..6109)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6263..6851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6343..6362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7022..7813,7814..7882))
                     /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     7645..7664
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7883..7987)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8062..8081)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag cttggtaccg agctcggatc cactagtcca gtgtggtgga attgcccttg
      721 aattcaccat ggactccact gcctgcttga agtccttgct cctgactgtc agtcagtaca
      781 aagccgtgaa gtcagaggcg aacgccactc agcttttgcg gcacttggag gtaatttctg
      841 gacagaaact cacacgacta tttacatcaa atcagatatt aacaagtgaa tgcttgagtt
      901 gccttgtaga gctacttgaa gaccccaaca taagtgcttc actgatctta agtattatcg
      961 gtttgctgtc tcaactagca gtagacattg aaaccagaga ttgtcttcag aatacatata
     1021 atctgaatag tgtgctggcg ggagtggttt gtcggagcag ccacactgat tcggtgtttt
     1081 tgcagtgcat tcaacttcta cagaagttaa catataatgt caaaattttc tattctggtg
     1141 ccaatataga tgaattaatt acgttcctga tagatcacat tcaatcttct gaagatgagt
     1201 taaaaatgcc ttgtctagga ttattggcaa atctttgtcg gcacaatctt tctgttcaaa
     1261 cgcacataaa gacattgagt aatgtgaaat ctttttatcg aactcttatc accttgttgg
     1321 cccatagtag tttaactgtg gttgtgtttg cactttcaat attatccagt ttgacattaa
     1381 atgaagaggt gggggaaaag ctattccatg ctcgaaacat tcatcagact tttcaactaa
     1441 tatttaatat tctcataaac ggtgatggca ctctaactag aaagtattca gttgacctac
     1501 tgatggatct ccttaagaat cctaaaattg ctgattatct caccagatat gagcactttt
     1561 cttcatgtct tcaccaagta ttaggtcttc ttaatggaaa ggatcctgat tcctcttcaa
     1621 aggttttaga attacttctt gccttctgtt cagtgactca gctgcgccat atgctcactc
     1681 agatgatgtt tgaacagtct ccacctggca gcgccactct gggaagccat actaaatgtt
     1741 tagaacctac tgtggctcta ctgcgctggt taagccaacc tttggacgga tcagaaaact
     1801 gttctgtttt agcattggag ttgttcaagg aaatatttga ggatgtcata gatgctgcta
     1861 actgttcctc ggctgatcgt tttgtgaccc ttctgctgcc tacaatcctt gatcaacttc
     1921 agttcacaga acaaaatcta gatgaggctt taacaagaaa aaaatgtgaa aggattgcca
     1981 aggccattga agttttgtta actctctgtg gagatgatac actaaaaatg catattgcaa
     2041 aaatcttgac aactgtcaag tgtaccactc ttatagaaca acaatttaca tatggcaaga
     2101 ttgacctggg atttggaaca aaggttgcag attctgaatt atgcaaactt gctgctgatg
     2161 taattttgaa aactcttgat ttgattaaca aacttaaacc attggttcct ggtatggaag
     2221 taagcttcta caaaatactt caggacccac gtttgattac tcctttggct tttgctttaa
     2281 cgtcagataa tagagaacaa gtacagtctg gactgagaat attagaggag gctgctccac
     2341 tgccagattt tcctgcttta gtacttggag aaagtatagc agcaaacaat gcctatagac
     2401 aacaggaaac agaacatata cccagaaaaa tgccctggca atcatcaaat cacagttttc
     2461 caacatcaat aaagtgttta actcctcatt tgaaagatgg tgttcctgga ttgaatattg
     2521 aagaattaat agagaaactt cagtctggaa tggtggtaaa ggatcagatt tgtgatgtga
     2581 gaatatctga cataatggat gtatatgaaa tgaaactatc cacattagct tccaaagaaa
     2641 gcaggctaca agatcttttg gaaacaaaag ctctagccct tgcacaggct gatagactga
     2701 ttgctcagca tcgctgtcaa agaactcaag ctgaaacaga ggcacggaca cttgctagta
     2761 tgttgagaga agttgagaga aaaaatgaag agcttagtgt gttgctgaag gcgcagcaag
     2821 ttgaatcaga aagagcgcag agtgatattg agcatctctt tcaacataat aggaagttag
     2881 agtctgtggc tgaagaacat gaaatactga caaaatccta catggaactt cttcagagaa
     2941 atgaaagtac tgaaaagaag aataaagatt tacagatcac atgtgattct ctgaataaac
     3001 aaattgagac agtgaaaaag ttgaatgagt cactcaagga acaaaatgaa aaaagtattg
     3061 cccaattaat agagaaagaa gaacagagaa aagaagtaca gaatcagcta gtagacagag
     3121 aacataagct agcaaatttg catcaaaaaa caaaagtaca agaagaaaag attaaaacct
     3181 tacaaaagga aagggaagat aaggaagaaa ccattgatat ccttagaaaa gaattaagca
     3241 gaacagaaca gataagaaaa gagttgagca ttaaggcttc ctccctagag gttcaaaagg
     3301 cacaattaga aggtcgtttg gaagagaaag agtccttggt gaaacttcag caagaggaat
     3361 tgaacaaaca ctcccacatg atagcaatga tccacagttt aagtggtgga aaaataaatc
     3421 cagaaactgt gaatctcagt ataggtacca agggcaattc tgcagatatc cagcacagtg
     3481 gcggccgctc gagtctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     3541 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
     3601 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3661 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3721 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3781 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3841 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3901 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3961 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     4021 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4081 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4141 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4201 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     4261 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     4321 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4381 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4441 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4501 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4561 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4621 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4681 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4741 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4801 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4861 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4921 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4981 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     5041 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     5101 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     5161 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     5221 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     5281 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     5341 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5401 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5461 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5521 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5581 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5641 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5701 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5761 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5821 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5881 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5941 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     6001 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     6061 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     6121 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     6181 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     6241 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     6301 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6361 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6421 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     6481 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6541 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6601 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6661 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6721 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6781 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6841 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6901 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6961 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     7021 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     7081 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     7141 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     7201 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     7261 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     7321 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7381 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7441 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7501 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7561 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7621 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7681 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7741 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7801 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7861 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7921 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7981 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     8041 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     8101 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     8161 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     8221 gcttcgcgat gtacgggcca gatatacgcg tt
//
