LOCUS       sequence-482396-        8443 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Over-express Sc rad24.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8443)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 8443)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Sc [rad24 + RFC5]-pLANT/RIL
FEATURES             Location/Qualifiers
     source          1..8443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        231..249
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    250..274
                     /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)."
     RBS             289..311
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     terminator      1490..1537
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      complement(1751..1798)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     RBS             3905..3927
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3942..3966
                     /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(3967..3985)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     tRNA            4414..4490
                     /gene="argU"
                     /product="E. coli tRNA-Arg recognizing AGA and AGG"
                     /label=argU
     CDS             complement(4944..5759)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4944..5759)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5667..5686
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5751..5770
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6241..6786
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     6687..6706
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6903..6920
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
       61 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
      121 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
      181 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat taatacgact
      241 cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt tgtttaactt
      301 taagaaggag atatacatat gtcattgtgg gtagataaat acagacctaa gtccttgaat
      361 gctctttcac ataatgaaga gttgacaaat tttctaaaat cgttatctga tcagcctcgt
      421 gatttacctc atcttttact gtatggacca aatggtacag gtaagaaaac gcgttgtatg
      481 gcattattgg agtccatatt tggacctgga gtctatagat tgaaaattga tgtcagacaa
      541 tttgtcactg cttcgaacag aaaactagaa ctgaatgtgg tcagctcgcc ataccattta
      601 gagatcacgc caagtgatat gggtaacaat gatagaattg tcatccaaga actattgaaa
      661 gaagtggctc aaatggaaca agtggacttt caagattcta aggatggact tgcccataga
      721 tataagtgtg ttattatcaa cgaggcgaac tcgttaacaa aagatgctca agctgcttta
      781 agacgtacca tggaaaaata ctccaaaaac attaggttga taatggtctg cgattcgatg
      841 tcgcctataa ttgctcctat caaatcccgt tgtctgttga ttcgttgtcc tgcaccaagc
      901 gatagcgaaa tttcaactat cttgtctgat gtggtgacaa atgaaagaat acaactagaa
      961 acaaaggata ttttaaaaag aattgctcag gcatcgaatg gaaacttgcg agtctcccta
     1021 ttaatgcttg aatctatggc actgaacaac gaattagcat tgaaaagcag tagccctata
     1081 ataaaaccag attggattat agtgatccat aaattaacga ggaaaatcgt taaagagaga
     1141 tctgtcaatt ctttaatcga atgcagagct gtcctatacg atttactagc tcattgtata
     1201 cctgccaata tcatcttaaa ggaactaacg ttttctttgt tggatgtgga aaccctgaat
     1261 accacgaata aatcgtccat aattgaatac tcaagtgttt ttgacgaaag attatcactt
     1321 ggaaacaaag caatattcca tttggaaggg ttcatagcaa aagttatgtg ctgtctagat
     1381 taatgtaaga tatgtcataa atactgtata agtcacacaa aaagcggatc cggctgctaa
     1441 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     1501 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     1561 atcgataagc tttaatgcgg tagtttatca cagttaaatt gctaacgcag tcaggcaccg
     1621 tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct ggatgctgta
     1681 ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatccg gatatagttc
     1741 ctcctttcag caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctagtt
     1801 attgctcagc ggtggcagca gccaactcag cttcctttcg ggctttgtta gcagccggat
     1861 catagatttg tgtggatatt tcctggggtt ttctcgtcaa atttaaagag taaaaagtta
     1921 gagtatttcc agatctgaat ctgaaaggga ctcactgata actggcgctt tacgcggctg
     1981 tctctgggac agaatttcac atagagactc gtcttcgtcg cttctaccaa atgtttcatt
     2041 gccaatatta ttgctgttat cgctatcgct atcgacaata gggtcatcat taaaagaagt
     2101 ttcttcgtct tcaagatcgt cgttagccat catcacattt ctttcatatt ggtcaatcaa
     2161 catgcgaagc cttctgtccc tttccttttt ttgatcctct aaatggtcac aattattgct
     2221 atcattatcc atcaatggtg ctagtccatc ctccacagat agtgcttcga taggcccgcc
     2281 tatccgatcc acaacgtcga ttccgttctc aactttcatt atatcactaa atttgtccat
     2341 ggtttgtttg ggccccgagg agccactcgg aagattcttc aaataataga gaatgtattt
     2401 ttttttataa ctctgacact ttctaattag aggtgcgtag tagccaaatt ctagagttat
     2461 attcctgaaa gaatgtaccg cgttgtactt ataaagacta acatttaacc aatcttcagc
     2521 ttgaacttta aatgaatttt gtaattttct tactttccat tctcttggaa aataaaccgt
     2581 tccatgatta tggccttgtt tactgatgtt acgaaaggtt ttgcgcactt ctcgtaaacc
     2641 atactcattg gattctggta gaccattcat attatcacac tcgctcaggc aatccacaat
     2701 tgatgatgca tcagaaatgc tgaattcgcc tttattaaat gtgttatagt tctctaatat
     2761 tcctaatttg aaatcctctt tcgataacaa attgttcgaa ttttcgaaga ggttattaat
     2821 catttcgtta tcgttattcg tgctatggga accatgtatc accttcccaa tggcatgaaa
     2881 gtatgatatg gtggattctc gggttgaaat cggcaaactt ccacttgatg tcgcccaaaa
     2941 ttgaagggtc gtaatggccg acctaatatc accagtctct tgcgcaatat aatctatgac
     3001 ttcctgtctt ttattccatt tatttttctc cttcaacatt ttcatattct gtacacaaat
     3061 aaatttcaag tgctttttta ataaagtgct gttaattgga ttaaacttaa ttcttttcaa
     3121 ccttggatgc atcaatattt ctttgttcat tatggtttct gcactaaatg tataatcaat
     3181 accaaatttg cgataattat tatcgttctc tggaatttca cattcagtta tacatataac
     3241 aaggggaggt aatagaggct ccgaactata tagccactgt aatataagtt gttgaaatcg
     3301 acgtctggta tctatatgga agacgttggg aaggtcctcg ataagtatta atgacaggtt
     3361 ggacatcaca agataccgtg cgccttttaa gaactcacta aagctttcca tctgaggaag
     3421 atcgttgact atacaatcac ctctaaactc ggtcacttta tgctcgttcg gggtgcttcg
     3481 aaaggacgtt ccgttgctgt tttgtctgta tttaggaact aagatttttg agagttcttt
     3541 tatgaccgta cttttagagc atccactggg gccagacagt agtaggatcc tatgcttggc
     3601 gttaggtaaa aacatggcat ctaaagcttc ttgtacatct ttaagttttc ttttatgtat
     3661 ggccacttgc tccaaacaat tgggcttgaa cttttcgtac cactgttcac cgccgtcgtc
     3721 gttgatactt gggaggatac tagacgtatc ttgcttggaa agaaagtcat tttcagtgct
     3781 tctcgcctta cgaactggcg aagtcggtct agatgagctc cattttgtta tttgcgagcc
     3841 caatgaactg agactatatt gtaataaggg ccgtttgttc aaattcgtac tatccatatg
     3901 tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac
     3961 aattccccta tagtgagtcg tattaatttc gcgggatcga gatctcgatc ctctacgccg
     4021 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
     4081 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
     4141 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgatcccgc
     4201 aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg
     4261 ccgaggatgc tgtacaattg acgacgacaa gggcccgggc aaactagtaa tcagacgcgg
     4321 tcgttcactt gttcagcaac cagatcaaaa gccattgact cagcaagggt tgaccgtata
     4381 attcacgcga ttacaccgca ttgcggtatc aacgcgccct tagctcagtt ggatagagca
     4441 acgaccttct aagtcgtggg ccgcaggttc gaatcctgca gggcgcgcca ttacaattca
     4501 atcagttacg ccttctttat atcctccagc catggccttg aaatggcgtt agtcatgaaa
     4561 tatagaccgc catcgagtac cccttgtacc cttaactctt cctgatacgt aaataatgat
     4621 ttggtggccc ttgctggact tgaaccagcg accaagcgat tatgagtcgc ctgctctaac
     4681 cactgagcta aagggccttg agtgtgcaat aacaatactt ataaaccacg caataaacat
     4741 gatgatctag agaatcccgt cgtagccacc atcttttttt gcgggagtgg cgaaattggt
     4801 agacgcacca gatttaggtt ctggcgccgc taggtgtgcg agttcaagtc tcgcctcccg
     4861 caccattcac cagaaagcgt tgatcggatg ccctcgagtc gggcagcgtt gggtcctggc
     4921 cacgggtgcg catgatcgaa ttcttagaaa aactcatcga gcatcaaatg aaactgcaat
     4981 ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga
     5041 gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg
     5101 actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt
     5161 gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct
     5221 ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc
     5281 aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa
     5341 ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca
     5401 atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc
     5461 gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga
     5521 ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg
     5581 ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag
     5641 attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca
     5701 tccatgttgg aatttaatcg cggcctcgag caagacgttt cccgttgaat atggctcata
     5761 acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga ccaaaatccc
     5821 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5881 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5941 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6001 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     6061 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cacgatgata
     6121 agctgtcaaa catgagaatt acaacttata tcgtatgggg ctgacttcag gtgctacatt
     6181 tgaagagata aattgcactg aaatctagaa atattttatc tgattaataa gatgatcttc
     6241 ttgagatcgt tttggtctgc gcgtaatctc ttgctctgaa aacgaaaaaa ccgccttgca
     6301 gggcggtttt tcgaaggttc tctgagctac caactctttg aaccgaggta actggcttgg
     6361 aggagcgcag tcaccaaaac ttgtcctttc agtttagcct taaccggcgc atgacttcaa
     6421 gactaactcc tctaaatcaa ttaccagtgg ctgctgccag tggtgctttt gcatgtcttt
     6481 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcggac tgaacggggg
     6541 gttcgtgcat acagtccagc ttggagcgaa ctgcctaccc ggaactgagt gtcaggcgtg
     6601 gaatgagaca aacgcggcca taacagcgga atgacaccgg taaaccgaaa ggcaggaaca
     6661 ggagagcgca cgagggagcc gccaggggga aacgcctggt atctttatag tcctgtcggg
     6721 tttcgccacc actgatttga gcgtcagatt tcgtgatgct tgtcaggggg gcggagccta
     6781 tggaaaaacg gctttgccgc ggccctctca cttccctgtt aagtatcttc ctggcatctt
     6841 ccaggaaatc tccgccccgt tcgtaagcca tttccgctcg ccgcagtcga acgaccgagc
     6901 gtagcgagtc agtgagcgag gaagcggaat atatcctgta tcacatattc tgctgacgca
     6961 ccggtgcagc cttttttctc ctgccacatg aagcgattca cagatgtctg cctgttcatc
     7021 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     7081 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     7141 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     7201 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     7261 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     7321 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     7381 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     7441 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     7501 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     7561 cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct
     7621 gctggagatg gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt
     7681 tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg
     7741 ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg
     7801 cagacaaggt atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag
     7861 gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc
     7921 gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc
     7981 tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc
     8041 cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg
     8101 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     8161 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     8221 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     8281 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     8341 ctgactgggt tgaaggctct caagggcatc ggtcgacgct ctcccttatg cgactcctgc
     8401 attaggaagc agcccagtag taggttgagg ccgttgagca ccg
//
