LOCUS       sequence-439558-        7877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression vector of Flag-tagged CIP2A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7877)
  AUTHORS   Adam S, Rossi SE, Moatti N, De Marco Zompit M, Xue Y, Ng TF, 
            Alvarez-Quilon A, Desjardins J, Bhaskaran V, Martino G, Setiaputra 
            D, Noordermeer SM, Ohsumi TK, Hustedt N, Szilard RK, Chaudhary N, 
            Munro M, Veloso A, Melo H, Yin SY, Papp R, Young JTF, Zinda M, 
            Stucki M, Durocher D
  TITLE     The CIP2A-TOPBP1 axis safeguards chromosome stability and is a 
            synthetic lethal target for BRCA-mutated cancer.
  JOURNAL   Nat Cancer. 2021 Dec;2(12):1357-1371. doi: 
            10.1038/s43018-021-00266-w. Epub 2021 Nov 11.
  PUBMED    35121901
REFERENCE   2  (bases 1 to 7877)
  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/s43018-021-00266-w"; journalName: "Nat Cancer"; date: 
            "2021-12"; volume: "2"; issue: "12"; pages: "1357-1371"
COMMENT     Sequence Label: pcDNA5-FRT/T0-Flag-CIP2A
FEATURES             Location/Qualifiers
     source          1..7877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(67..171)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(246..265)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        437..816
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        817..1020
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     971..991
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    1022..1040
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1043..1061
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1071..1095
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1195..1218
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4031..4048)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4037..4261
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4394..4413)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4545..4592
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             4600..5622
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5752..5873
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5789..5808)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5843..5862
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5922..5938)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5922..5938)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5935..5957)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5946..5962
                     /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(5970..5976,5977..5994,5995..6000))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6015..6036
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6153..6170)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6324..6912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6404..6423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7083..7874,7875..7877,1..66))
                     /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     7706..7725
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
       61 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
      121 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
      181 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
      241 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
      301 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
      361 gacaattgca tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg
      421 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
      481 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      541 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      601 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      661 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      721 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      781 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      841 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      901 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      961 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
     1021 ctccctatca gtgatagaga tctccctatc agtgatagag atcgtcgacg agctcgttta
     1081 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
     1141 cgggaccgat ccagcctccg gactctagcg tttaaactta agcttggtac catggactac
     1201 aaagacgatg acgataaagc cgccagttta aacggcgcgc caatggactc cactgcctgc
     1261 ttgaagtcct tgctcctgac tgtcagtcag tacaaagccg tgaagtcaga ggcgaacgcc
     1321 actcagcttt tgcggcactt ggaggtaatt tctggacaga aactcacacg actatttaca
     1381 tcaaatcaga tattaacaag tgaatgcttg agttgccttg tagagctact tgaagacccc
     1441 aacataagtg cttcactgat cttaagtatt atcggtttgc tgtctcaact agcagtagac
     1501 attgaaacca gagattgtct tcagaataca tataatctga atagtgtgct ggcgggagtg
     1561 gtttgtcgga gcagccacac tgattcggtg tttttgcagt gcattcaact tctacagaag
     1621 ttaacatata atgtcaaaat tttctattct ggtgccaata tagatgaatt aattacgttc
     1681 ctgatagatc acattcaatc ttctgaagat gagttaaaaa tgccttgtct aggattattg
     1741 gcaaatcttt gtcggcacaa tctttctgtt caaacgcaca taaagacatt gagtaatgtg
     1801 aaatcttttt atcgaactct tatcaccttg ttggcccata gtagtttaac tgtggttgtg
     1861 tttgcacttt caatattatc cagtttgaca ttaaatgaag aggtggggga aaagctattc
     1921 catgctcgaa acattcatca gacttttcaa ctaatattta atattctcat aaacggtgat
     1981 ggcactctaa ctagaaagta ttcagttgac ctactgatgg atctccttaa gaatcctaaa
     2041 attgctgatt atctcaccag atatgagcac ttttcttcat gtcttcacca agtattaggt
     2101 cttcttaatg gaaaggatcc tgattcctct tcaaaggttt tagaattact tcttgccttc
     2161 tgttcagtga ctcagctgcg ccatatgctc actcagatga tgtttgaaca gtctccacct
     2221 ggcagcgcca ctctgggaag ccatactaaa tgtttagaac ctactgtggc tctactgcgc
     2281 tggttaagcc aacctttgga cggatcagaa aactgttctg ttttagcatt ggagttgttc
     2341 aaggaaatat ttgaggatgt catagatgct gctaactgtt cctcggctga tcgttttgtg
     2401 acccttctgc tgcctacaat ccttgatcaa cttcagttca cagaacaaaa tctagatgag
     2461 gctttaacaa gaaaaaaatg tgaaaggatt gccaaggcca ttgaagtttt gttaactctc
     2521 tgtggagatg atacactaaa aatgcatatt gcaaaaatct tgacaactgt caagtgtacc
     2581 actcttatag aacaacaatt tacatatggc aagattgacc tgggatttgg aacaaaggtt
     2641 gcagattctg aattatgcaa acttgctgct gatgtaattt tgaaaactct tgatttgatt
     2701 aacaaactta aaccattggt tcctggtatg gaagtaagct tctacaaaat acttcaggac
     2761 ccacgtttga ttactccttt ggcttttgct ttaacgtcag ataatagaga acaagtacag
     2821 tctggactga gaatattatt ggaggctgct ccactgccag attttcctgc tttagtactt
     2881 ggagaaagta tagcagcaaa caatgcctat agacaacagg aaacagaaca tatacccaga
     2941 aaaatgccct ggcaatcatc aaatcacagt tttccaacat caataaagtg tttaactcct
     3001 catttgaaag atggtgttcc tggattgaat attgaagaat taatagagaa acttcagtct
     3061 ggaatggtgg taaaggatca gatttgtgat gtgagaatat ctgacataat ggatgtatat
     3121 gaaatgaaac tatccacatt agcttccaaa gaaagcaggc tacaagatct tttggaaaca
     3181 aaagctctag cccttgcaca ggctgataga ctgattgctc agcatcgctg tcaaagaact
     3241 caagctgaaa cagaggcacg gacacttgct agtatgttga gagaagttga gagaaaaaat
     3301 gaagagctta gtgtgttgct gaaggcgcag caagttgaat cagaaagagc gcagagtgat
     3361 attgagcatc tctttcaaca taataggaag ttagagtctg tggctgaaga acatgaaata
     3421 ctgacaaaat cctacatgga acttcttcag agaaatgaaa gtactgaaaa gaagaataaa
     3481 gatttacaga tcacatgtga ttctctgaat aaacaaattg agacagtgaa aaagttgaat
     3541 gagtcactca aggaacaaaa tgaaaaaagt attgcccaat taatagagaa agaagaacag
     3601 agaaaagaag tacagaatca gctagtagac agagaacata agctagcaaa tttgcatcaa
     3661 aaaacaaaag tacaagaaga aaagattaaa accttacaaa aggaaaggga agataaggaa
     3721 gaaaccattg atatccttag aaaagaatta agcagaacag aacagataag aaaagagttg
     3781 agcattaagg cttcctccct agaggttcaa aaggcacaat tagaaggtcg tttggaagag
     3841 aaagagtcct tggtgaaact tcagcaagag gaattgaaca aacactccca catgatagca
     3901 atgatccaca gtttaagtgg tggaaaaata aatccagaaa ctgtgaatct cagtatataa
     3961 gaattctgca gatatccatc acactggcgg ccgctcgagt ctagagggcc cgtttaaacc
     4021 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     4081 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     4141 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     4201 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     4261 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     4321 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4381 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4441 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     4501 ctcgacccca aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga
     4561 agttcctatt ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg
     4621 acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc
     4681 tcggagggcg aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg
     4741 cgggtaaata gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca
     4801 tcggccgcgc tcccgattcc ggaagtgctt gacattgggg agttcagcga gagcctgacc
     4861 tattgcatct cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg
     4921 cccgctgttc tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc
     4981 cagacgagcg ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt
     5041 gatttcatat gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac
     5101 accgtcagtg cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc
     5161 cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat
     5221 ggccgcataa cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag
     5281 gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac
     5341 ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc
     5401 attggtcttg accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg
     5461 gcgcagggtc gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa
     5521 atcgcccgca gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt
     5581 ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc
     5641 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5701 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     5761 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5821 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5881 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     5941 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     6001 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     6061 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6121 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     6181 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     6241 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     6301 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6361 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6421 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6481 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6541 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6601 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6661 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6721 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6781 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6841 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6901 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6961 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     7021 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     7081 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     7141 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     7201 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     7261 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     7321 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     7381 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     7441 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     7501 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     7561 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     7621 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     7681 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     7741 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     7801 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     7861 agcgtttctg ggtgagc
//
