LOCUS       sequence-315915-        7357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express HA-tagged POLE2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7357)
  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 7357)
  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: pMXS-IRES-Blast HA-POLE2
FEATURES             Location/Qualifiers
     source          1..7357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..417
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     347..369
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     385..401
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    427..801
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     739..755
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             867..956
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             complement(2287..2298)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     misc_feature    join(2616..3190,3191..3193,3194..3202)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2783..2800)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3010..3029
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3203..3598
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             3769..4238
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4368..4385)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4539..5127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4619..4638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5291..6082,6083..6151))
                     /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     5914..5933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6152..6256)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             6286..6879
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    6942..7299
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
       61 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
      121 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
      181 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
      241 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
      301 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
      361 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccccc
      421 atatggccat atgagatctt atatggggca cccccgcccc ttgtaaactt ccctgaccct
      481 gacatgacaa gagttactaa cagcccctct ctccaagctc acttacaggc tctctactta
      541 gtccagcacg aagtctggag acctctggcg gcagcctacc aagaacaact ggaccgaccg
      601 gtggtacctc acccttaccg agtcggcgac acagtgtggg tccgccgaca ccagactaag
      661 aacctagaac ctcgctggaa aggaccttac acagtcctgc tgaccacccc caccgccctc
      721 aaagtagacg gcatcgcagc ttggatacac gccgcccacg tgaaggctgc cgaccccggg
      781 ggtggaccat cctctagact gccggatcta gctagttaat taaggatccc agtgtggtgg
      841 tacgggaatt caccatggct gctgtatatc cttatgatgt ccctgattac gcgggatacc
      901 cgtacgatgt tcccgactat gcgggaagct acccttacga cgtaccggat tatgcccccg
      961 cggctggagg cggaggaggc ggtatggcgc ctgagagatt gagatctcgc gcgctcagtg
     1021 cgttcaaatt gcgaggcttg ctcttgaggg gcgaggcgat caagtatctt acagaggcac
     1081 tgcaatctat ttcagaactg gagctggagg acaaactcga gaagattatt aatgctgtgg
     1141 aaaaacagcc cttgagttct aatatgattg aaaggtcagt agtagaagcc gcagtacagg
     1201 agtgctccca gagcgtggat gaaacgatag agcacgtgtt caatattata ggagcatttg
     1261 acatcccgcg ctttgtatac aattctgagc ggaaaaaatt cctgccgctc cttatgacta
     1321 accatcccgc acctaacctg tttggcacac cccgcgataa ggcggaaatg tttcgagagc
     1381 gatacacaat tctccaccag cgaacgcata gacatgagct gtttacaccg cctgtcatcg
     1441 ggtcacaccc cgacgaaagt ggtagcaagt ttcaactgaa aactatagaa acgctgcttg
     1501 gctccactac aaagatcggc gatgctatag tgcttggaat gataacccaa ctgaaagaag
     1561 gaaaattttt tctcgaggat ccgactggta ctgtgcagct cgacctctcc aaggcgcagt
     1621 tccactctgg gctgtataca gaggcgtgct tcgtccttgc tgaaggatgg tttgaggatc
     1681 aggtcttcca cgtaaatgct tttgggttcc cgccaaccga acccagcagc acaactcggg
     1741 cgtattacgg aaatatcaac ttctttgggg gtcctagcaa cacaagcgtc aagacttccg
     1801 ctaaattgaa gcagctcgag gaggagaata aggacgccat gtttgtcttc ttgtctgatg
     1861 tttggctgga tcaggttgag gtactggaga aacttcgcat tatgttcgcg gggtattccc
     1921 cggcgccgcc cacatgtttc attctctgtg gtaatttttc atctgcccct tatggtaaaa
     1981 accaggtgca ggcgctcaag gattccctca agactcttgc cgatataatt tgtgaatatc
     2041 ccgatattca ccagagctcc agatttgtgt tcgtcccagg ccctgaagat ccgggatttg
     2101 ggtcaatcct gcctaggccg ccacttgcag aatccatcac aaatgagttc cgccaacgcg
     2161 tgcctttttc cgtgttcacg acgaatcctt gccggattca atactgcaca caggagatca
     2221 cagttttcag agaggatctc gtcaacaaga tgtgtagaaa ttgtgtgcgc ttcccgagct
     2281 ccaatcttgc aattcccaat cacttcgtca aaactatatt gtcccaaggc catcttacac
     2341 cgttgccttt gtatgtctgc ccggtgtatt gggcttacga ttatgctctt cgggtatacc
     2401 cggtaccgga cctccttgtg attgctgaca aatacgaccc tttcactaca accaatacgg
     2461 aatgcctctg cattaaccct ggtagctttc cacgcagtgg attcagtttt aaggtcttct
     2521 acccctctaa taaaactgtg gaggatagca aacttcaagg tttttgagtc gagggccggc
     2581 gcgccgcggc cgctacgtaa attccgcccc ccccccccct ctccctcccc cccccctaac
     2641 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     2701 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     2761 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     2821 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     2881 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     2941 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     3001 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     3061 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     3121 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     3181 cgatgataat atggccacaa ccatggttcc tttgtctcaa gaagaatcca ccctcattga
     3241 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     3301 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     3361 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     3421 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     3481 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     3541 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagt
     3601 cgacgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     3661 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
     3721 aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc
     3781 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
     3841 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     3901 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
     3961 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     4021 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     4081 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
     4141 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     4201 ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta
     4261 atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc
     4321 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     4381 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     4441 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     4501 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4561 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4621 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4681 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4741 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4801 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4861 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4921 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4981 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5041 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5101 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5161 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5221 ttcacctaga tccttttgcg gccggccgca aatcaatcta aagtatatat gagtaaactt
     5281 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5341 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5401 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5461 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5521 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5581 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5641 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5701 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5761 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5821 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5881 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5941 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6001 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6061 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6121 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6181 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6241 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcataatga aagaccccac
     6301 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     6361 actgagaata gaaaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     6421 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     6481 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     6541 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     6601 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     6661 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     6721 acccctcact cggcgcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     6781 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     6841 tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg
     6901 gagacccctg cccagggacc accgacccac caccgggagg taagctggcc agcaacttat
     6961 ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag
     7021 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt cggaacaccc
     7081 ggccgcaacc ctgggagacg tcccagggac ttcgggggcc gtttttgtgg cccgacctga
     7141 gtccaaaaat cccgatcgtt ttggactctt tggtgcaccc ccctaatagg agggatatgt
     7201 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
     7261 ggtttgggac cgaagccgcg ccgcgcgtct tgtctgctgc agcatcgttc tgtgttgtct
     7321 ctgtctgact gtgtttctgt atttgtctga aaattag
//
