LOCUS       sequence-320626-        7322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Suppress CDC25A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7322)
  AUTHORS   Sviderskiy VO, Blumenberg L, Gorodetsky E, Karakousi TR, Hirsh N, 
            Alvarez SW, Terzi EM, Kaparos E, Whiten GC, Ssebyala S, Tonzi P, Mir
            H, Neel BG, Huang TT, Adams S, Ruggles KV, Possemato R
  TITLE     Hyperactive CDK2 Activity in Basal-like Breast Cancer Imposes a 
            Genome Integrity Liability that Can Be Exploited by Targeting DNA 
            Polymerase epsilon.
  JOURNAL   Mol Cell. 2020 Oct 28. pii: S1097-2765(20)30723-1. doi: 
            10.1016/j.molcel.2020.10.016.
  PUBMED    33152268
REFERENCE   2  (bases 1 to 7322)
  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: "Mol Cell. 
            2020 Oct 28. pii: S1097-2765(20)30723-1. doi: 
            10.1016/j.molcel.2020.10.016."
COMMENT     Sequence Label: pLKO.1B CDC25A_2
FEATURES             Location/Qualifiers
     source          1..7322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    26..259
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             444..488
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     promoter        666..914
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     666..686
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     837..856
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    1011..1128
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1177..1687
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     1606..1624
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             1699..2094
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     misc_feature    2128..2716
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2181..2201)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2599..2610)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2788..3021
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    3099..3220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3136..3155)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3190..3209
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3260..3395
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3415..3434)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3416..3434)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3444..3461)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3444..3460)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3453..3475)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3602..4057
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3689..3708)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3899..3920
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4083..4187
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4188..4256,4257..5048)
                     /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     complement(4406..4425)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5219..5807
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5708..5727
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5961..5978
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6095..6116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6131..6136,6137..6154,6155..6161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6169..6185
                     /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)."
     primer_bind     6174..6196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6193..6209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6193..6209
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6228..6248
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6230..6248
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6276..6502
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6503..6683
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6730..6855
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
ORIGIN
        1 agagagaaaa aagagcagtg ggaataggag ctttgttcct tgggttcttg ggagcagcag
       61 gaagcactat gggcgcagcg tcaatgacgc tgacggtaca ggccagacaa ttattgtctg
      121 gtatagtgca gcagcagaac aatttgctga gggctattga ggcgcaacag catctgttgc
      181 aactcacagt ctggggcatc aagcagctcc aggcaagaat cctggctgtg gaaagatacc
      241 taaaggatca acagctcctg gggatttggg gttgctctgg aaaactcatt tgcaccactg
      301 ctgtgccttg gaatgctagt tggagtaata aatctctgga acagatttgg aatcacacga
      361 cctggatgga gtgggacaga gaaattaaca attacacaag cttaatacac tccttaattg
      421 aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt attggaatta gataaatggg
      481 caagtttgtg gaattggttt aacataacaa attggctgtg gtatataaaa ttattcataa
      541 tgatagtagg aggcttggta ggtttaagaa tagtttttgc tgtactttct atagtgaata
      601 gagttaggca gggatattca ccattatcgt ttcagaccca cctcccaacc ccgaggggac
      661 ccagagaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta
      721 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac
      781 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact
      841 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga
      901 aaggacgagg taccggagat cgcctgggta atgaatactc gagtattcat tacccaggcg
      961 atcttttttg aattctagat cttgagacaa atggcagtat tcatccacaa ttttaaaaga
     1021 aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac
     1081 atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac
     1141 agggacagca gagatccact ttggcgccgg ctcgaggggg cccgggttgc gccttttcca
     1201 aggcagccct gggtttgcgc agggacgcgg ctgctctggg cgtggttccg ggaaacgcag
     1261 cggcgccgac cctgggtctc gcacattctt cacgtccgtt cgcagcgtca cccggatctt
     1321 cgccgctacc cttgtgggcc ccccggcgac gcttcctgct ccgcccctaa gtcgggaagg
     1381 ttccttgcgg ttcgcggcgt gccggacgtg acaaacggaa gccgcacgtc tcactagtac
     1441 cctcgcagac ggacagcgcc agggagcaat ggcagcgcgc cgaccgcgat gggctgtggc
     1501 caatagcggc tgctcagcag ggcgcgccga gagcagcggc cgggaagggg cggtgcggga
     1561 ggcggggtgt ggggcggtag tgtgggccct gttcctgccc gcgcggtgtt ccgcattctg
     1621 caagcctccg gagcgcacgt cggcagtcgg ctccctcgtt gaccgaatca ccgacctctc
     1681 tccccagggg gatccaccat ggccaagcct ttgtctcaag aagaatccac cctcattgaa
     1741 agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca
     1801 gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga
     1861 ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact
     1921 tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga
     1981 caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag
     2041 ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggcctgcag
     2101 ctgcagtagt aaacgcgtta agtcgacaat caacctctgg attacaaaat ttgtgaaaga
     2161 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
     2221 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
     2281 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
     2341 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
     2401 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
     2461 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
     2521 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
     2581 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
     2641 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
     2701 tgggccgcct ccccgcgtcg actttaagac caatgactta caaggcagct gtagatctta
     2761 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag
     2821 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     2881 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     2941 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     3001 cagtgtggaa aatctctagc agtacgtata gtagttcatg tcatcttatt attcagtatt
     3061 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     3121 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3181 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     3241 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3301 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     3361 attccagaag tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat
     3421 agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     3481 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     3541 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     3601 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3661 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3721 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     3781 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     3841 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     3901 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     3961 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     4021 aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg
     4081 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4141 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     4201 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     4261 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     4321 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     4381 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     4441 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     4501 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4561 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     4621 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4681 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4741 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4801 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4861 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4921 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4981 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     5041 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     5101 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5161 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5221 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5281 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5341 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5401 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5461 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5521 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5581 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5641 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5701 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5761 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5821 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5881 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     5941 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     6001 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     6061 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     6121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     6181 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
     6241 actaaaggga acaaaagctg gagctgcaag cttaatgtag tcttatgcaa tactcttgta
     6301 gtcttgcaac atggtaacga tgagttagca acatgcctta caaggagaga aaaagcaccg
     6361 tgcatgccga ttggtggaag taaggtggta cgatcgtgcc ttattaggaa ggcaacagac
     6421 gggtctgaca tggattggac gaaccactga attgccgcat tgcagagata ttgtatttaa
     6481 gtgcctagct cgatacataa acgggtctct ctggttagac cagatctgag cctgggagct
     6541 ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt gagtgcttca
     6601 agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca gaccctttta
     6661 gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc gaaagggaaa
     6721 ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc aagaggcgag
     6781 gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa ggagagagat
     6841 gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga aaaaattcgg
     6901 ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc aagcagggag
     6961 ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg tagacaaata
     7021 ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc attatataat
     7081 acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac caaggaagct
     7141 ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca agcggccgct
     7201 gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat tatataaata
     7261 taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga gaagagtggt
     7321 gc
//
