LOCUS       sequence-322383-       10068 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express POLA1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10068)
  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 10068)
  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 POLA1
FEATURES             Location/Qualifiers
     source          1..10068
                     /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    5333..5900
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5350..5902
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5494..5511)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5721..5740
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5914..6309
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MVPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTGV
                     NVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGIK
                     AIVKDSDGQPTAVGIRELLPSGYVWEG"
     LTR             6480..6949
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7079..7096)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7250..7838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7330..7349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8002..8793,8794..8862))
                     /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     8625..8644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8863..8967)
                     /gene="bla"
                     /label=AmpR promoter
     LTR             8997..9590
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    9653..10010
                     /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 ttgtaaagct agtcccctga ccttcaccgt
     4861 gcagctgtcc aataaactga taatggacat ccgcagattt atcaagaaat actatgacgg
     4921 ctggcttata tgtgaagaac caacgtgcag gaaccgcact aggcacctgc ctcttcagtt
     4981 cagcaggacc ggacctctgt gtcctgcatg tatgaaagca actctgcagc ccgagtacag
     5041 cgataagtca ctgtataccc aactttgttt ctataggtac atttttgatg cagaatgtgc
     5101 tttggaaaaa ctgacaaccg atcatgaaaa ggacaagttg aagaagcagt tttttacacc
     5161 aaaggttctc caagattacc ggaagctgaa gaatactgct gaacagtttc tgtctaggtc
     5221 cggctacagc gaagtcaacc tgtccaaatt gtttgccggg tgcgcagtta agagttaaga
     5281 attcggccgg cgcgccgcgg ccgctacgta aattccgccc ccccccccnn tctccctccc
     5341 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     5401 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     5461 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     5521 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     5581 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     5641 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     5701 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     5761 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     5821 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     5881 tttgaaaaac acgatgataa tatggccaca accatggttc ctttgtctca agaagaatcc
     5941 accctcattg aaagagcaac ggctacaatc aacagcatcc ccatctctga agactacagc
     6001 gtcgccagcg cagctctctc tagcgacggc cgcatcttca ctggtgtcaa tgtatatcat
     6061 tttactgggg gaccttgtgc agaactcgtg gtgctgggca ctgctgctgc tgcggcagct
     6121 ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca ggggcatctt gagcccctgc
     6181 ggacggtgcc gacaggtgct tctcgatctg catcctggga tcaaagccat agtgaaggac
     6241 agtgatggac agccgacggc agttgggatt cgtgaattgc tgccctctgg ttatgtgtgg
     6301 gagggctaag tcgacgataa aataaaagat tttatttagt ctccagaaaa aggggggaat
     6361 gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt gcaaggcatg
     6421 gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag atggaacagc
     6481 tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
     6541 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     6601 cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc agtttctaga
     6661 gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg ccttatttga
     6721 actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc gagctcaata
     6781 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     6841 acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt
     6901 gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac atgcagcatg
     6961 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
     7021 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7081 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7141 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7201 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7261 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     7321 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     7381 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     7441 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     7501 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     7561 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     7621 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     7681 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     7741 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     7801 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     7861 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     7921 caaaaaggat cttcacctag atccttttgc ggccggccgc aaatcaatct aaagtatata
     7981 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     8041 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     8101 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     8161 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     8221 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     8281 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     8341 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     8401 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     8461 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     8521 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     8581 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     8641 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     8701 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     8761 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     8821 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     8881 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     8941 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcataatg
     9001 aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg
     9061 aaaaatacat aactgagaat agaaaagttc agatcaaggt caggaacaga tggaacagct
     9121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
     9181 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     9241 ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca gtttctagag
     9301 aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc cttatttgaa
     9361 ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg agctcaataa
     9421 aagagcccac aacccctcac tcggcgcgcc agtcctccga ttgactgagt cgcccgggta
     9481 cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg ctgttccttg
     9541 ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt tgggggctcg
     9601 tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag gtaagctggc
     9661 cagcaactta tctgtgtctg tccgattgtc tagtgtctat gactgatttt atgcgcctgc
     9721 gtcggtacta gttagctaac tagctctgta tctggcggac ccgtggtgga actgacgagt
     9781 tcggaacacc cggccgcaac cctgggagac gtcccaggga cttcgggggc cgtttttgtg
     9841 gcccgacctg agtccaaaaa tcccgatcgt tttggactct ttggtgcacc cccctaatag
     9901 gagggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa
     9961 tttttgcttt cggtttggga ccgaagccgc gccgcgcgtc ttgtctgctg cagcatcgtt
    10021 ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg aaaattag
//
