LOCUS       sequence-322394-        9077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express ISC mutant POLD1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9077)
  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 9077)
  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 ISCmut
FEATURES             Location/Qualifiers
     source          1..9077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    117..474
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             533..949
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     879..901
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     917..933
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    959..1333
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1271..1287
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1382..1447
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(2261..2272)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     misc_feature    join(4868..5442,5443..5445,5446..5454)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5035..5052)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5262..5281
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5455..5850
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             6021..6490
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6620..6637)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6791..7379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6871..6890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7543..8334,8335..8403))
                     /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     8166..8185
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8404..8508)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             join(8538..9077,1..54)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt catttggggg
       61 ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
      121 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga ttttatgcgc
      181 ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
      241 gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg gggccgtttt
      301 tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg caccccccta
      361 ataggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
      421 tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct gctgcagcat
      481 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac
      541 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
      601 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
      661 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
      721 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
      781 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
      841 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
      901 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccc ccatatggcc
      961 atatgagatc ttatatgggg cacccccgcc ccttgtaaac ttccctgacc ctgacatgac
     1021 aagagttact aacagcccct ctctccaagc tcacttacag gctctctact tagtccagca
     1081 cgaagtctgg agacctctgg cggcagccta ccaagaacaa ctggaccgac cggtggtacc
     1141 tcacccttac cgagtcggcg acacagtgtg ggtccgccga caccagacta agaacctaga
     1201 acctcgctgg aaaggacctt acacagtcct gctgaccacc cccaccgccc tcaaagtaga
     1261 cggcatcgca gcttggatac acgccgccca cgtgaaggct gccgaccccg ggggtggacc
     1321 atcctctaga ctgccggatc tagctagtta attaaggatc cctaaatcgc aattgaccat
     1381 ggactacaag gaccacgacg gtgactacaa ggaccacgac atcgactaca aggacgacga
     1441 cgacaaggga ggaggcggag gtggaatgga tggcaagcgg cggccaggcc cagggcccgg
     1501 ggtgccccca aagcgggccc gtgggggcct ctgggatgat gatgatgcac ctcggccatc
     1561 ccaattcgag gaggacctgg cactgatgga ggagatggag gcagaacaca ggctgcagga
     1621 gcaggaggag gaggagctgc agtcagtcct ggagggggtt gcagacgggc aggtcccacc
     1681 atcagccata gatcctcgct ggcttcggcc cacaccacca gcgctggacc cccagacaga
     1741 gcccctcatc ttccaacagt tggagattga ccattatgtg ggcccagcac agcctgtgcc
     1801 tggggggccc ccaccatccc acggctccgt gcctgtgctc cgcgccttcg gggtcaccga
     1861 tgaggggttc tctgtctgct gccacatcca cggcttcgct ccctacttct acaccccagc
     1921 gccccctggt ttcgggcccg agcacatggg tgacctgcaa cgggagctga acttggccat
     1981 cagccgggac agtcgcgggg ggagggagct gactgggccg gccgtgctgg ctgtggaact
     2041 gtgctcccga gagagcatgt ttgggtacca cgggcacggc ccctccccgt tcctgcgcat
     2101 caccgtggcg ctgccgcgcc tcgtggcccc ggcccgccgt ctcctggaac agggcatccg
     2161 tgtggcaggc ctgggcacgc ccagcttcgc gccctacgag gccaacgtcg actttgagat
     2221 ccggttcatg gtggacacgg acatcgtcgg ctgcaactgg ctggagctcc cagctgggaa
     2281 atacgccctg aggctgaagg agaaggctac gcagtgccag ctggaggcgg acgtgctgtg
     2341 gtctgacgtg gtcagtcacc caccggaagg gccatggcag cgcattgcgc ccttgcgcgt
     2401 gctcagcttc gatatcgagt gcgccggccg caaaggcatc ttccctgagc ctgagcggga
     2461 ccctgtcatc cagatctgct cgctgggcct gcgctggggg gagccggagc ccttcctacg
     2521 cctggcgctc accctgcggc cctgtgcccc catcctgggt gccaaggtgc agagctacga
     2581 gaaggaggag gacctgctgc aggcctggtc caccttcatc cgtatcatgg accccgacgt
     2641 gatcaccggt tacaacatcc agaacttcga ccttccgtac ctcatctctc gggcccagac
     2701 cctcaaggta caaacattcc ctttcctggg ccgtgtggcc ggcctttgct ccaacatccg
     2761 ggactcttca ttccagtcca agcagacggg ccggcgggac accaaggttg tcagcatggt
     2821 gggccgcgtg cagatggaca tgctgcaggt gctgctgcgg gagtacaagc tccgctccta
     2881 cacgctcaat gccgtgagct tccacttcct gggcgagcag aaggaggacg tgcagcacag
     2941 catcatcacc gacctgcaga atgggaacga ccagacccgc cgccgcctgg ctgtgtactg
     3001 cctgaaggat gcctacctgc cactgcggct gctggagcgg ctcatggtgc tggtgaacgc
     3061 cgtggagatg gcgagggtca ctggcgtgcc cctcagctac ctgctcagtc gtggccagca
     3121 ggtcaaggtc gtatcccagc tgttgcggca ggccatgcac gaggggctgc tgatgcccgt
     3181 ggtgaagtca gagggcggcg aggactacac gggagccact gtcatcgagc ccctcaaagg
     3241 gtactacgac gtccccatcg ccaccctgga cttctcctcg ctgtacccgt ccatcatgat
     3301 ggcccacaac ctgtgttaca ccacgctcct tcggcccggg actgcacaga aactgggcct
     3361 gactgaggat cagttcatca ggacccccac cggggacgag tttgtgaaga cctcagtgcg
     3421 gaaggggctg ctgccccaga tcctggagaa cctgctcagt gcccggaaga gggccaaggc
     3481 cgagctggcc aaggagacag accccctccg gcgccaggtc ctggatggac ggcagctggc
     3541 gctgaaggtg agcgccaact ccgtatacgg cttcactggc gcccaggtgg gcaagttgcc
     3601 gtgcctggag atctcacaga gcgtcacggg gttcggacgt cagatgatcg agaaaaccaa
     3661 gcagctggtg gagtctaagt acacagtgga gaatggctac agcaccagtg ccaaggtggt
     3721 gtatggtgac actgactccg tcatgtgccg attcggcgtg tcctcggtgg ctgaggcgat
     3781 ggccctgggg cgggaggccg cggactgggt gtcaggtcac ttcccgtcgc ccatccggct
     3841 ggagtttgag aaggtctact tcccatacct gcttatcagc aagaagcgct acgcgggcct
     3901 gctcttctcc tcccggcccg acgcccacga ccgcatggac tgcaagggcc tggaggccgt
     3961 gcgcagggac aactgccccc tcgtggccaa cctggtcact gcctcactgc gccgcctgct
     4021 catcgaccga gaccctgagg gcgcggtggc tcacgcacag gacgtcatct cggacctgct
     4081 gtgcaaccgc atcgatatca gccagctcgt gattactaaa gagctcacca gagccgcttc
     4141 agattacgcg ggaaaacaag cccatgtcga actggcagaa cggatgagaa aaagagatcc
     4201 agggagcgct ccctcactcg gggatagagt cccctatgta atcatttcag cagccaaggg
     4261 ggtagccgcc tacatgaaat cagaggaccc tttgttcgtg ctggaacaca gtcttcccat
     4321 tgatacacag tactacttgg aacagcagtt ggctaaaccg ctgctgcgga tctttgaacc
     4381 aattttgggc gaaggtaggg cagaggcagt attgctccgg ggtgatcata cgcgctgcaa
     4441 aacggtactc actggcaaag ttggtggcct gctcgcgttc gcaaagagac gaaactgctg
     4501 tatcgggtgt aggacagtgc tcagccatca gggagcagtc tgtgaatttt gccagcccag
     4561 ggagagcgag ctttatcaga aagaggtctc tcacctcaac gctctggagg agagattttc
     4621 cagactgtgg actcagagcc agcggagcca aggaagcttg cacgaggatg taattagcac
     4681 ttctcgcgat agccccatct tttatatgag aaaaaaggtt cggaaggatc tcgaagatca
     4741 agagcagctc ctcagacgat ttggccctcc tggacccgaa gcctggctcg agctaaatcg
     4801 gtttaaacgc gctaatgcgg ccgctacgta aattccgccc cccccccccc tctcccyccc
     4861 cccccccccc ctctccctcc ccccccccta acgttactgg ccgaagccgc ttggaataag
     4921 gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt ggcaatgtga
     4981 gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt tcccctctcg
     5041 ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt
     5101 gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca cctggcgaca
     5161 ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc
     5221 agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc tcaagcgtat
     5281 tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct gatctggggc
     5341 ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga
     5401 accacgggga cgtggttttc ctttgaaaaa cacgatgata atatggccac aaccatggtt
     5461 cctttgtctc aagaagaatc caccctcatt gaaagagcaa cggctacaat caacagcatc
     5521 cccatctctg aagactacag cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc
     5581 actggtgtca atgtatatca ttttactggg ggaccttgtg cagaactcgt ggtgctgggc
     5641 actgctgctg ctgcggcagc tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac
     5701 aggggcatct tgagcccctg cggacggtgc cgacaggtgc ttctcgatct gcatcctggg
     5761 atcaaagcca tagtgaagga cagtgatgga cagccgacgg cagttgggat tcgtgaattg
     5821 ctgccctctg gttatgtgtg ggagggctaa gtcgacgata aaataaaaga ttttatttag
     5881 tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc tagcttaagt
     5941 aacgccattt tgcaaggcat ggaaaaatac ataactgaga atagagaagt tcagatcaag
     6001 gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc
     6061 ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa acaggatatc
     6121 tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc
     6181 cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa
     6241 atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc
     6301 ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc
     6361 gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag ttgcatccga
     6421 cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac ccgtcagcgg
     6481 gggtctttca catgcagcat gtatcaaaat taatttggtt ttttttctta agtatttaca
     6541 ttaaatggcc atagttgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6601 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6661 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6721 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6781 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6841 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6901 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6961 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7021 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7081 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7141 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7201 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     7261 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7321 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7381 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7441 ggattttggt catgagatta tcaaaaagga tcttcaccta gatccttttg cggccggccg
     7501 caaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7561 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7621 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7681 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7741 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7801 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7861 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7921 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7981 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     8041 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     8101 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     8161 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     8221 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     8281 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     8341 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     8401 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     8461 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     8521 aaaagtgcca cctgcataat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     8581 acgccatttt gcaaggcatg gaaaaataca taactgagaa tagaaaagtt cagatcaagg
     8641 tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     8701 tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct
     8761 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     8821 agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     8881 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     8941 tctgctcccc gagctcaata aaagagccca caacccctca ctcggcgcgc cagtcctccg
     9001 attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     9061 ttgtggtctc gctgttc
//
