LOCUS       sequence-315907-       10054 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Express ISC mutant POLA1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10054)
  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 10054)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 POLA1 ISCmut
FEATURES             Location/Qualifiers
     source          1..10054
                     /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"
     misc_feature    join(5313..5887,5888..5890,5891..5899)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5480..5497)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5707..5726
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5900..6295
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             6466..6935
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7065..7082)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7236..7824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7316..7335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7988..8779,8780..8848))
                     /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     8611..8630
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8849..8953)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             8983..9576
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    9639..9996
                     /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 cctgcaggcc tcgaggctag catggctcct gtacacgggg atgattgtga
      901 aatcggagct tctgctttgt ccgatagtgg gtcatttgta tctagcaggg ccagaagaga
      961 aaaaaaaagc aaaaaaggcc ggcaagaggc tctggagcgg cttaagaagg ccaaagccgg
     1021 agaaaaatat aaatacgaag tggaggattt tactggtgtc tacgaggaag tggacgagga
     1081 gcagtactct aaattggtgc aagctcgaca ggacgatgac tggatcgtgg atgatgatgg
     1141 cattggctat gtcgaagatg gaagggagat cttcgatgat gatttggagg acgatgcact
     1201 tgacgcggat gaaaagggca aggatggaaa agctcggaac aaggataaaa gaaacgtgaa
     1261 gaagctggcc gtcactaagc ccaataatat taagagtatg tttatcgcct gtgcaggcaa
     1321 aaaaaccgct gataaggcag tggatttgag caaggatggc ctcttggggg acatactgca
     1381 ggacttgaac accgaaactc cccaaatcac tcctccgcct gttatgattc tcaaaaagaa
     1441 gcgaagcatc ggcgccagcc ctaatccttt cagcgttcat actgccactg cagtgccttc
     1501 cgggaagatc gcttcccccg tgagcagaaa ggaaccaccc ttgactccag tcccattgaa
     1561 gagagctgag tttgccggcg acgatgtaca ggttgagagt actgaggagg aacaggaatc
     1621 tggggcaatg gaatttgagg acggcgattt tgatgagcct atggaagtgg aagaggttga
     1681 tttggagcct atggccgcta aagcctggga caaggaaagc gaaccagccg aggaggttaa
     1741 acaagaagcc gacagtggca aagggacagt ttcttacctg ggctcattct tgcctgacgt
     1801 gagttgctgg gatattgacc aagagggcga ttcatcattt tcagtgcaag aagtgcaagt
     1861 ggacagctct cacctgcccc tcgttaaagg tgctgatgag gagcaggtct ttcatttcta
     1921 ttggctggat gcttatgagg atcagtacaa tcagcctggc gtggtattcc tctttggcaa
     1981 agtgtggatc gagagcgctg aaacccacgt gtcctgctgt gtgatggtga aaaatataga
     2041 gcgaactctc tacttcctcc caagagagat gaagattgat ctgaatactg ggaaggaaac
     2101 gggcaccccg atcagtatga aggacgtgta cgaagaattt gacgaaaaaa ttgccaccaa
     2161 gtacaagatc atgaagttta agtccaagcc cgttgagaag aattatgcat tcgagattcc
     2221 cgacgtacct gagaagtctg aatatctgga ggtgaagtat tccgctgaga tgcctcagct
     2281 cccacaggat ctgaaaggcg agactttttc acacgtcttt ggaactaata caagttccct
     2341 ggagctgttt cttatgaaca ggaagattaa ggggccctgt tggttggagg tcaagtctcc
     2401 ccagctcttg aaccaacccg tctcctggtg taaagtggag gcaatggccc tgaagccaga
     2461 tcttgtaaac gtaatcaagg acgtgagtcc cccgccactc gttgtcatgg ctttctctat
     2521 gaagaccatg caaaatgcta agaaccacca aaatgaaatt attgccatgg ccgctctcgt
     2581 gcatcattcc tttgccctgg ataaagcagc ccccaaaccc cccttccagt cccatttttg
     2641 tgtggtcagc aaaccaaaag actgtatttt cccttatgcc ttcaaagaag ttatagagaa
     2701 aaagaacgtg aaagtcgaag tggcggctac cgaacgcact ttgctgggct ttttcttggc
     2761 caaagtgcat aaaatcgacc ctgatattat tgtgggacat aatatctacg gtttcgaact
     2821 cgaagtcctc cttcagcgca tcaatgtgtg taaggccccg cattggtcta agattggcag
     2881 gctgaaacgg tcaaatatgc caaagctggg tgggcgatct ggatttggag agaggaacgc
     2941 cacgtgtggc cgcatgattt gcgatgtgga aatatccgcc aaagaattga ttagatgcaa
     3001 atcataccat cttagcgaac tcgtgcagca gattctcaaa acagagagag tcgtgatacc
     3061 tatggaaaac attcagaaca tgtacagtga gtcctctcag ctgctttacc tcctggaaca
     3121 cacttggaaa gacgctaagt tcattttgca gataatgtgc gagttgaatg tcctgccttt
     3181 ggccttgcaa atcaccaaca tagccggcaa tattatgagc aggacgctga tggggggtag
     3241 gtcagagcga aacgagttcc tgctgctgca cgctttttac gagaacaatt atattgtgcc
     3301 agacaagcag atattccgaa agccacagca gaagttgggc gacgaagacg aggagattga
     3361 tggggacacg aacaagtaca agaagggacg caagaaggca gcctacgcag gcggactcgt
     3421 gctggatcca aaggtcggat tctacgataa attcattctg ctgctcgact ttaactctct
     3481 ctatccttct atcatccaag agtttaacat atgctttaca actgtccaac gagtggcctc
     3541 tgaggcgcag aaggtgacag aggacggcga acaagagcag atccctgaac tgcccgatcc
     3601 atcactggag atggggatat tgcctaggga aattcggaaa cttgtggaaa gaagaaagca
     3661 agttaaacag ctgatgaaac agcaagacct caacccagat ctgatcttgc aatatgatat
     3721 tagacagaag gctctgaagc tcacagcgaa ctcaatgtac ggatgcctcg gtttttccta
     3781 cagcagattt tatgcaaaac ctttggcagc cctcgtaacc tataaaggta gggaaatact
     3841 catgcacacc aaggagatgg tacagaaaat gaatcttgag gtgatttacg gggataccga
     3901 tagtattatg atcaatacca actccacgaa tttggaggag gtttttaagc tcggtaacaa
     3961 agtcaaaagc gaggtaaaca aactgtacaa actgctggag atcgacatag acggagtatt
     4021 caagtccctg cttctgctga agaagaagaa atatgctgcc ctcgtggtcg aaccaacctc
     4081 cgatggcaac tatgtgacca agcaagagct caaaggtctg gacattgtgc gaagagactg
     4141 gtgcgacctg gccaaggata ccgggaattt tgtaatcggt cagatactga gcgatcagtc
     4201 tcgggatact atcgtggaaa acattcaaaa gaggctgatc gaaattggag agaatgtgct
     4261 gaatggctcc gtgccagtgt cacaattcga gattaacaag gcattgacaa aggacccaca
     4321 ggattaccca gataaaaaga gcctccctca tgtgcacgtc gccctctgga tcaatagcca
     4381 aggagggagg aaagttaaag ccggggacac cgtgtcatat gtgatctgcc aggacgggtc
     4441 taacctcacg gcatcccaga gggcctatgc tcctgagcag ctccagaagc aggacaacct
     4501 cacgatagac acgcagtact atctggctca gcagatccac cctgtagtgg ccagaatctg
     4561 tgaaccaatt gatggaatag acgcggttct tattgcaact tggctcgggc tcgatcctac
     4621 acagttccgg gtgcatcatt accataagga cgaagaaaat gatgccttgc tgggggggcc
     4681 tgcccaactt acagacgagg agaaataccg agattgtgag agatttaagt gtccctgtcc
     4741 cacctgcggt accgaaaaca tctatgacaa tgtgtttgat ggctccggaa cagacatgga
     4801 gcccagcctg tatagatgtt ctaatatcga ttgtaaagca tcacctttga cctttaccgt
     4861 gcaacttagc aataagctga ttatggacat aagaagattc ataaaaaaat actatgatgg
     4921 atggttgatt tcagaagaac cgacctcacg caaccgcacg agacacctcc cgcttcagtt
     4981 ctcacggacg ggaccgttgt caccagcgtc aatgaaggcc acgctgcagc ccgagtactc
     5041 agacaagagc ctttatacac agctctgttt ttatagatat atttttgacg cggagtgtgc
     5101 gctggagaag ctgaccaccg atcacgaaaa ggacaagttg aaaaaacagt ttttcactcc
     5161 aaaggtgctg caggactacc gcaagctcaa aaacacggcg gagcagttcc tgtcacgctc
     5221 cgggtactct gaagtgaacc tctcaaaact gtttgcgggg tgcgcagtaa aaagctaaag
     5281 atctgcggcc gctacgtaaa ttccnncccc cccccctctc cctccccccc ccctaacgtt
     5341 actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc
     5401 atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc
     5461 attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag
     5521 gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg
     5581 cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat
     5641 acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga
     5701 gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc
     5761 cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg
     5821 ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga
     5881 tgataatatg gccacaacca tggttccttt gtctcaagaa gaatccaccc tcattgaaag
     5941 agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg ccagcgcagc
     6001 tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta ctgggggacc
     6061 ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca acctgacttg
     6121 tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac ggtgccgaca
     6181 ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg atggacagcc
     6241 gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg gctaagtcga
     6301 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     6361 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     6421 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
     6481 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     6541 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     6601 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     6661 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     6721 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     6781 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     6841 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     6901 tgagtgattg actacccgtc agcgggggtc tttcacatgc agcatgtatc aaaattaatt
     6961 tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg aatcggccaa
     7021 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     7081 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     7141 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     7201 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7261 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7321 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7381 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7441 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     7501 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     7561 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     7621 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     7681 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     7741 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     7801 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     7861 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     7921 acctagatcc ttttgcggcc ggccgcaaat caatctaaag tatatatgag taaacttggt
     7981 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     8041 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     8101 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     8161 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     8221 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     8281 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     8341 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     8401 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8461 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8521 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8581 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8641 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8701 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8761 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8821 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8881 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8941 taggggttcc gcgcacattt ccccgaaaag tgccacctgc ataatgaaag accccacctg
     9001 taggtttggc aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact
     9061 gagaatagaa aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa
     9121 caggatatct gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc
     9181 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     9241 aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt
     9301 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
     9361 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
     9421 cctcactcgg cgcgccagtc ctccgattga ctgagtcgcc cgggtacccg tgtatccaat
     9481 aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt tccttgggag ggtctcctct
     9541 gagtgattga ctacccgtca gcgggggtct ttcatttggg ggctcgtccg ggatcgggag
     9601 acccctgccc agggaccacc gacccaccac cgggaggtaa gctggccagc aacttatctg
     9661 tgtctgtccg attgtctagt gtctatgact gattttatgc gcctgcgtcg gtactagtta
     9721 gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttcgg aacacccggc
     9781 cgcaaccctg ggagacgtcc cagggacttc gggggccgtt tttgtggccc gacctgagtc
     9841 caaaaatccc gatcgttttg gactctttgg tgcacccccc taataggagg gatatgtggt
     9901 tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt
     9961 ttgggaccga agccgcgccg cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg
    10021 tctgactgtg tttctgtatt tgtctgaaaa ttag
//
