LOCUS       sequence-405352-        4056 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A plasmid that facilitates production of mRNA encoding V5 epitope 
            tagged catalytically dead I-PpoI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4056)
  AUTHORS   van Sluis M, McStay B
  TITLE     A localized nucleolar DNA damage response facilitates recruitment of
            the homology-directed repair machinery independent of cell cycle 
            stage.
  JOURNAL   Genes Dev. 2015 Jun 1;29(11):1151-63. doi: 10.1101/gad.260703.115. 
            Epub 2015 May 27.
  PUBMED    26019174
REFERENCE   2  (bases 1 to 4056)
  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"; doi: 
            "10.1101/gad.260703.115"; journalName: "Genes Dev"; date: 
            "2015-06-1- 1"; volume: "29"; issue: "11"; pages: "1151-63"
COMMENT     Sequence Label: pBstSK+IRES-V5-IPpoI (mutant)
FEATURES             Location/Qualifiers
     source          1..4056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     669..685
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    705..1271
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    721..1273
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(865..882)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1092..1111
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1288..1329
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1868..1888)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1868..1886)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1907..1923)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1907..1923)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1920..1942)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1931..1947
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1955..1961,1962..1979,1980..1985))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2000..2021
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2138..2155)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2309..2897)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2389..2408)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3068..3859,3860..3928))
                     /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     3691..3710
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3929..4033)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattccgcc ccccccccta
      721 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
      781 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
      841 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
      901 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
      961 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     1021 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     1081 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     1141 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     1201 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     1261 cacgatgata atatggccac aaccatgggt aagcctatcc ctaaccctct cctcggtctc
     1321 gattctacgc gcatggcgct caccaatgct caaatcttgg ctgtgattga cagctgggaa
     1381 gaaacagtcg gtcagtttcc agtgataacg caccatgtac cattaggtgg cggtctgcaa
     1441 ggaacgctcc attgttacga gatcccccta gcagctcctt atggggttgg ctttgctaag
     1501 aatgggccta cccgctggca atacaaacgg acaatcaatc aagtcgtcca cagatgggga
     1561 tcccacacag tcccttttct attagaaccg gataacatca acggcaaaac ctgcacagca
     1621 tcggcgctat gtcataatac tcgatgccac aatcccttgc acttgtgctg ggagtcacta
     1681 gacgacaaca aaggcagaaa ctggtgcccg ggtcccaacg ggggatgtgt ccatgcggtg
     1741 gtttgtttaa ggcagggtcc gttgtacggc ccgggggcga ctgtggcagg tcctcaacaa
     1801 aggggcagtc actttgtggt ataaatctag agcggccgcc accgcggtgg agctccagct
     1861 tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca tagctgtttc
     1921 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1981 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2041 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2101 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2161 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2221 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2281 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2341 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2401 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2461 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2521 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2581 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2641 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2701 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2761 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2821 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2881 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2941 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3001 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3061 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3121 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3181 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3241 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3301 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3361 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3421 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3481 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3541 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3601 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3661 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3721 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3781 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3841 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3901 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3961 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4021 taggggttcc gcgcacattt ccccgaaaag tgccac
//
