LOCUS       sequence-322925-        9056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express POLD1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9056)
  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 9056)
  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 POLD1
FEATURES             Location/Qualifiers
     source          1..9056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             42..458
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     388..410
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     426..442
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    468..842
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     780..796
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             891..956
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(1770..1781)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     misc_feature    join(4356..4930,4931..4933,4934..4942)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4523..4540)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4750..4769
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4943..5338
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             5509..5978
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6108..6125)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6279..6867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6359..6378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7031..7822,7823..7891))
                     /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     7654..7673
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7892..7996)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             8026..8619
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    8682..9039
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
       61 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
      121 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
      181 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
      241 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
      301 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
      361 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
      421 cctcccttta tccagccctc actccttctc taggcgcccc catatggcca tatgagatct
      481 tatatggggc acccccgccc cttgtaaact tccctgaccc tgacatgaca agagttacta
      541 acagcccctc tctccaagct cacttacagg ctctctactt agtccagcac gaagtctgga
      601 gacctctggc ggcagcctac caagaacaac tggaccgacc ggtggtacct cacccttacc
      661 gagtcggcga cacagtgtgg gtccgccgac accagactaa gaacctagaa cctcgctgga
      721 aaggacctta cacagtcctg ctgaccaccc ccaccgccct caaagtagac ggcatcgcag
      781 cttggataca cgccgcccac gtgaaggctg ccgaccccgg gggtggacca tcctctagac
      841 tgccggatct agctagttaa ttaaggatcc ctaaatcgca attgaccatg gactacaagg
      901 accacgacgg tgactacaag gaccacgaca tcgactacaa ggacgacgac gacaagggag
      961 gaggcggagg tggaatggat ggcaagcggc ggccaggccc agggcccggg gtgcccccaa
     1021 agcgggcccg tgggggcctc tgggatgatg atgatgcacc tcggccatcc caattcgagg
     1081 aggacctggc actgatggag gagatggagg cagaacacag gctgcaggag caggaggagg
     1141 aggagctgca gtcagtcctg gagggggttg cagacgggca ggtcccacca tcagccatag
     1201 atcctcgctg gcttcggccc acaccaccag cgctggaccc ccagacagag cccctcatct
     1261 tccaacagtt ggagattgac cattatgtgg gcccagcaca gcctgtgcct ggggggcccc
     1321 caccatccca cggctccgtg cctgtgctcc gcgccttcgg ggtcaccgat gaggggttct
     1381 ctgtctgctg ccacatccac ggcttcgctc cctacttcta caccccagcg ccccctggtt
     1441 tcgggcccga gcacatgggt gacctgcaac gggagctgaa cttggccatc agccgggaca
     1501 gtcgcggggg gagggagctg actgggccgg ccgtgctggc tgtggaactg tgctcccgag
     1561 agagcatgtt tgggtaccac gggcacggcc cctccccgtt cctgcgcatc accgtggcgc
     1621 tgccgcgcct cgtggccccg gcccgccgtc tcctggaaca gggcatccgt gtggcaggcc
     1681 tgggcacgcc cagcttcgcg ccctacgagg ccaacgtcga ctttgagatc cggttcatgg
     1741 tggacacgga catcgtcggc tgcaactggc tggagctccc agctgggaaa tacgccctga
     1801 ggctgaagga gaaggctacg cagtgccagc tggaggcgga cgtgctgtgg tctgacgtgg
     1861 tcagtcaccc accggaaggg ccatggcagc gcattgcgcc cttgcgcgtg ctcagcttcg
     1921 atatcgagtg cgccggccgc aaaggcatct tccctgagcc tgagcgggac cctgtcatcc
     1981 agatctgctc gctgggcctg cgctgggggg agccggagcc cttcctacgc ctggcgctca
     2041 ccctgcggcc ctgtgccccc atcctgggtg ccaaggtgca gagctacgag aaggaggagg
     2101 acctgctgca ggcctggtcc accttcatcc gtatcatgga ccccgacgtg atcaccggtt
     2161 acaacatcca gaacttcgac cttccgtacc tcatctctcg ggcccagacc ctcaaggtac
     2221 aaacattccc tttcctgggc cgtgtggccg gcctttgctc caacatccgg gactcttcat
     2281 tccagtccaa gcagacgggc cggcgggaca ccaaggttgt cagcatggtg ggccgcgtgc
     2341 agatggacat gctgcaggtg ctgctgcggg agtacaagct ccgctcctac acgctcaatg
     2401 ccgtgagctt ccacttcctg ggcgagcaga aggaggacgt gcagcacagc atcatcaccg
     2461 acctgcagaa tgggaacgac cagacccgcc gccgcctggc tgtgtactgc ctgaaggatg
     2521 cctacctgcc actgcggctg ctggagcggc tcatggtgct ggtgaacgcc gtggagatgg
     2581 cgagggtcac tggcgtgccc ctcagctacc tgctcagtcg tggccagcag gtcaaggtcg
     2641 tatcccagct gttgcggcag gccatgcacg aggggctgct gatgcccgtg gtgaagtcag
     2701 agggcggcga ggactacacg ggagccactg tcatcgagcc cctcaaaggg tactacgacg
     2761 tccccatcgc caccctggac ttctcctcgc tgtacccgtc catcatgatg gcccacaacc
     2821 tgtgttacac cacgctcctt cggcccggga ctgcacagaa actgggcctg actgaggatc
     2881 agttcatcag gacccccacc ggggacgagt ttgtgaagac ctcagtgcgg aaggggctgc
     2941 tgccccagat cctggagaac ctgctcagtg cccggaagag ggccaaggcc gagctggcca
     3001 aggagacaga ccccctccgg cgccaggtcc tggatggacg gcagctggcg ctgaaggtga
     3061 gcgccaactc cgtatacggc ttcactggcg cccaggtggg caagttgccg tgcctggaga
     3121 tctcacagag cgtcacgggg ttcggacgtc agatgatcga gaaaaccaag cagctggtgg
     3181 agtctaagta cacagtggag aatggctaca gcaccagtgc caaggtggtg tatggtgaca
     3241 ctgactccgt catgtgccga ttcggcgtgt cctcggtggc tgaggcgatg gccctggggc
     3301 gggaggccgc ggactgggtg tcaggtcact tcccgtcgcc catccggctg gagtttgaga
     3361 aggtctactt cccatacctg cttatcagca agaagcgcta cgcgggcctg ctcttctcct
     3421 cccggcccga cgcccacgac cgcatggact gcaagggcct ggaggccgtg cgcagggaca
     3481 actgccccct cgtggccaac ctggtcactg cctcactgcg ccgcctgctc atcgaccgag
     3541 accctgaggg cgcggtggct cacgcacagg acgtcatctc ggacctgctg tgcaaccgca
     3601 tcgatatcag ccagctcgtg attactaaag agctcaccag agccgcttca gattacgcgg
     3661 gaaaacaagc ccatgtcgaa ctggcagaac ggatgagaaa aagagatcca gggagcgctc
     3721 cctcactcgg ggatagagtc ccctatgtaa tcatttcagc agccaagggg gtagccgcct
     3781 acatgaaatc agaggaccct ttgttcgtgc tggaacacag tcttcccatt gatacacagt
     3841 actacttgga acagcagttg gctaaaccgc tgctgcggat ctttgaacca attttgggcg
     3901 aaggtagggc agaggcagta ttgctccggg gtgatcatac gcgctgcaaa acggtactca
     3961 ctggcaaagt tggtggcctg ctcgcgttcg caaagagacg aaactgctgt atcgggtgta
     4021 ggacagtgct cagccatcag ggagcagtct gtgaattttg ccagcccagg gagagcgagc
     4081 tttatcagaa agaggtctct cacctcaacg ctctggagga gagattttcc agactgtgga
     4141 ctcagtgtca gcggtgtcaa ggaagcttgc acgaggatgt aatttgtact tctcgcgatt
     4201 gccccatctt ttatatgaga aaaaaggttc ggaaggatct cgaagatcaa gagcagctcc
     4261 tcagacgatt tggccctcct ggacccgaag cctggctcga gctaaatcgg tttaaacgcg
     4321 ctaatgcggc cgctacgtaa attccgcccc ccccccccct ctccctcccc cccccctaac
     4381 gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat gttattttcc
     4441 accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt cttcttgacg
     4501 agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt gaatgtcgtg
     4561 aaggaagcag ttcctctgga agcttcttga agacaaacaa cgtctgtagc gaccctttgc
     4621 aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc acgtgtataa
     4681 gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat agttgtggaa
     4741 agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc ccagaaggta
     4801 ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg tgtttagtcg
     4861 aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca
     4921 cgatgataat atggccacaa ccatggttcc tttgtctcaa gaagaatcca ccctcattga
     4981 aagagcaacg gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc
     5041 agctctctct agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg
     5101 accttgtgca gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac
     5161 ttgtatcgtc gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg
     5221 acaggtgctt ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca
     5281 gccgacggca gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaagt
     5341 cgacgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     5401 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaaatacat
     5461 aactgagaat agagaagttc agatcaaggt caggaacaga tggaacagct gaatatgggc
     5521 caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga acagatggaa
     5581 cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc ggctcagggc
     5641 caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag aaccatcaga
     5701 tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa ctaaccaatc
     5761 agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa aagagcccac
     5821 aacccctcac tcggggcgcc agtcctccga ttgactgagt cgcccgggta cccgtgtatc
     5881 caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg ggagggtctc
     5941 ctctgagtga ttgactaccc gtcagcgggg gtctttcaca tgcagcatgt atcaaaatta
     6001 atttggtttt ttttcttaag tatttacatt aaatggccat agttgcatta atgaatcggc
     6061 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     6121 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     6181 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     6241 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6301 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6361 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6421 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6481 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6541 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6601 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6661 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6721 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6781 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     6841 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     6901 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     6961 ttcacctaga tccttttgcg gccggccgca aatcaatcta aagtatatat gagtaaactt
     7021 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7081 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7141 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7201 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7261 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7321 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7381 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7441 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7501 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7561 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7621 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7681 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7741 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7801 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7861 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7921 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7981 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgcataatga aagaccccac
     8041 ctgtaggttt ggcaagctag cttaagtaac gccattttgc aaggcatgga aaaatacata
     8101 actgagaata gaaaagttca gatcaaggtc aggaacagat ggaacagctg aatatgggcc
     8161 aaacaggata tctgtggtaa gcagttcctg ccccggctca gggccaagaa cagatggaac
     8221 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     8281 aagaacagat ggtccccaga tgcggtccag ccctcagcag tttctagaga accatcagat
     8341 gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac taaccaatca
     8401 gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa agagcccaca
     8461 acccctcact cggcgcgcca gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc
     8521 aataaaccct cttgcagttg catccgactt gtggtctcgc tgttccttgg gagggtctcc
     8581 tctgagtgat tgactacccg tcagcggggg tctttcattt gggggctcgt ccgggatcgg
     8641 gagacccctg cccagggacc accgacccac caccgggagg taagctggcc agcaacttat
     8701 ctgtgtctgt ccgattgtct agtgtctatg actgatttta tgcgcctgcg tcggtactag
     8761 ttagctaact agctctgtat ctggcggacc cgtggtggaa ctgacgagtt cggaacaccc
     8821 ggccgcaacc ctgggagacg tcccagggac ttcgggggcc gtttttgtgg cccgacctga
     8881 gtccaaaaat cccgatcgtt ttggactctt tggtgcaccc ccctaatagg agggatatgt
     8941 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
     9001 ggtttgggac cgaagccgcg ccgcgcgtct tgtctgctgc agcatcgttc tgtgtt
//
