LOCUS       sequence-229316-        7080 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7080)
  AUTHORS   Matsuoka S, Ballif BA, Smogorzewska A, McDonald ER 3rd, Hurov KE, 
            Luo J, Bakalarski CE, Zhao Z, Solimini N, Lerenthal Y, Shiloh Y, 
            Gygi SP, Elledge SJ
  TITLE     ATM and ATR substrate analysis reveals extensive protein networks 
            responsive to DNA damage.
  JOURNAL   Science. 2007 May 25;316(5828):1160-6.
  PUBMED    17525332
REFERENCE   2  (bases 1 to 7080)
  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: "Science."; 
            date: "2007-05-25"; volume: "316(5828)"; pages: "1160-6"
COMMENT     Sequence Label: pBABE HA Chk2 KD, dominant negative chk2
FEATURES             Location/Qualifiers
     source          1..7080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             9..478
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    543..742
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             943..1359
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1289..1311
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1327..1343
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1400..1426
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1406..1432
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     primer_bind     complement(3411..3431)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3416..3745
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3596..3731
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3755..4354
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3755..3774)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4251..4271
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4575..5044
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5174..5191)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5345..5933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5425..5444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6093..6884,6885..6953))
                     /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     6716..6735
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6954..7058)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctgcagcct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
       61 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      121 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca
      181 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
      241 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
      301 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
      361 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
      421 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcatt
      481 tgggggctcg tccgggatcg ggagacccct gcccagggac caccgaccca ccaccgggag
      541 gcaagctggc cagcaactta tctgtgtctg tccgattgtc tagtgtctat gactgatttt
      601 atgcgcctgc gtcggtacta gttagctaac tagctctgta tctggcggac ccgtggtgga
      661 actgacgagt tctgaacacc cggccgcaac cctgggagac gtcccaggga ctttgggggc
      721 cgtttttgtg gcccgacctg aggaagggag tcgatgtgga atccgacccc gtcaggatat
      781 gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga atttttgctt
      841 tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat cgttctgtgt
      901 tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac tgttaccact
      961 cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca caaccagtcg
     1021 gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc aacctttaac
     1081 gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt taagatcaag
     1141 gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt gacctgggaa
     1201 gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa gcctccgcct
     1261 cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac cccgcctcga
     1321 tcctcccttt atccagccct cactccttct ctaggcgccg gccggatccc agtgtggtgg
     1381 tacgtaggaa ttcatggctt acccatacga tgttccagat tacgctagct tgggtggtca
     1441 tatggccatg gaggccccat ccatcacact ggcggccaat tccatgtctc gggagtcgga
     1501 tgttgaggct cagcagtctc atggcagcag tgcctgttca cagccccatg gcagcgttac
     1561 ccagtcccaa ggctcctcct cacagtccca gggcatatcc agctcctcta ccagcacgat
     1621 gccaaactcc agccagtcct ctcactccag ctctgggaca ctgagctcct tagagacagt
     1681 gtccactcag gaactctatt ctattcctga ggaccaagaa cctgaggacc aagaacctga
     1741 ggagcctacc cctgccccct gggctcgatt atgggccctt caggatggat ttgccaatct
     1801 tgaatgtgtg aatgacaact actggtttgg gagggacaaa agctgtgaat attgctttga
     1861 tgaaccactg ctgaaaagaa cagataaata ccgaacatac agcaagaaac actttcggat
     1921 tttcagggaa gtgggtccta aaaactctta cattgcatac atagaagatc acagtggcaa
     1981 tggaaccttt gtaaatacag agcttgtagg gaaaggaaaa cgccgtcctt tgaataacaa
     2041 ttctgaaatt gcactgtcac taagcagaaa taaagttttt gtcttttttg atctgactgt
     2101 agatgatcag tcagtttatc ctaaggcatt aagagatgaa tacatcatgt caaaaactct
     2161 tggaagtggt gcctgtggag aggtaaagct ggctttcgag aggaaaacat gtaagaaagt
     2221 agccataaag atcatcagca aaaggaagtt tgctattggt tcagcaagag aggcagaccc
     2281 agctctcaat gttgaaacag aaatagaaat tttgaaaaag ctaaatcatc cttgcatcat
     2341 caagattaaa aacttttttg atgcagaaga ttattatatt gttttggaat tgatggaagg
     2401 gggagagctg tttgacaaag tggtggggaa taaacgcctg aaagaagcta cctgcaagct
     2461 ctatttttac cagatgctct tggctgtgca gtaccttcat gaaaacggta ttatacaccg
     2521 tgccttaaag ccagagaatg ttttactgtc atctcaagaa gaggactgtc ttataaagat
     2581 tactgatttt gggcactcca agattttggg agagacctct ctcatgagaa ccttatgtgg
     2641 aacccccacc tacttggcgc ctgaagttct tgtttctgtt gggactgctg ggtataaccg
     2701 tgctgtggac tgctggagtt taggagttat tctttttatc tgccttagtg ggtatccacc
     2761 tttctctgag cataggactc aagtgtcact gaaggatcag atcaccagtg gaaaatacaa
     2821 cttcattcct gaagtctggg cagaagtctc agagaaagct ctggaccttg tcaagaagtt
     2881 gttggtagtg gatccaaagg cacgttttac gacagaagaa gccttaagac acccgtggct
     2941 tcaggatgaa gacatgaaga gaaagtttca agatcttctg tctgaggaaa atgaatccac
     3001 agctctaccc caggttctag cccagccttc tactagtcga aagcggcccc gtgaagggga
     3061 agccgagggt gccgagacca caaagcgccc agctgtgtgt gctgctgtgt tgtgaactcc
     3121 gtggtttgaa cacgaaagaa atgtaccttc tttcactctg tcatctttct tttctttgag
     3181 tctgtttttt tatagtttgt attttaatta tgggaataat tgctttttca cagtcactga
     3241 tgtacaatta aaaacctgat ggaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3301 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     3361 aaaaaaaaaa aaaaaaaaaa aggccttcgt ggcctcgagg tcgaccctgt ggaatgtgtg
     3421 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3481 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3541 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3601 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3661 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3721 ttttggaggc ctaggctttt gcaaaaagct taccatgacc gagtacaagc ccacggtgcg
     3781 cctcgccacc cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga
     3841 ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct
     3901 gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga
     3961 cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc
     4021 cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat
     4081 ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg
     4141 cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga
     4201 ggcggccgag cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc
     4261 cttctacgag cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg
     4321 cacctggtgc atgacccgca agcccggtgc ctgacgcccg ccccacgacc cgcagcgccc
     4381 gaccgaaagg agcgcacgac cccatgcatc gataaaataa aagattttat ttagtctcca
     4441 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
     4501 attttgcaag gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg
     4561 aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc
     4621 cggctcaggg ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt
     4681 aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc
     4741 tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc
     4801 ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc
     4861 tccccgagct caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga
     4921 ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg
     4981 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct
     5041 ttcacatgca gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat
     5101 ggccatagtt gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     5161 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     5221 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5281 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5341 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5401 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5461 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5521 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5581 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5641 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5701 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5761 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5821 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5881 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5941 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6001 tggtcatgag attatcaaaa aggatcttca cctagatcct tttgcggccg caaatcaatc
     6061 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6121 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6181 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6241 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6301 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6361 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6421 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6481 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6541 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6601 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6661 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     6721 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     6781 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     6841 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     6901 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     6961 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7021 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
//
