LOCUS       sequence-482357-        7106 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Over-express FLAG-rad24 in yeast (integrated).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7106)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 7106)
  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 FLAG-rad24-pRS404/GAL-L
FEATURES             Location/Qualifiers
     source          1..7106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(347..367)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(347..365)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(386..402)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(386..402)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(399..421)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    410..426
                     /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(434..440,441..458,459..464))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    479..500
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(617..634)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(788..1376)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(868..887)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1547..2338,2339..2407))
                     /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     2170..2189
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2408..2512)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2580..2598
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2636..2658)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2758..2777
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2798..3079
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             3080..3754
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     rep_origin      complement(3857..4312)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3994..4015)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4206..4225
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4438..4460
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4452..4469
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4453..4469
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4479..4498
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4479..4497
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     4522..4538
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4523..4539
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        4567..5231
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    4783..4900
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5195..5218
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             5244..5309
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
ORIGIN
        1 cttgaagacg aagaaacttc ttttaatgat gaccctattg tcgatagcga tagcgataac
       61 agcaataata ttggcaatga aacatttggt agaagcgacg aagacgagtc tctatgtgaa
      121 attctgtccc agagacagcc gcgtaaagcg ccagttatca gtgagtccct ttcagattca
      181 gatctggaaa tactctaact ttttactctt taaatttgac gagaaaaccc caggaaatat
      241 ccacacaaat ctatgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
      301 ccaccgctga gcggccgcca ccgcggtgga gctccagctt ttgttccctt tagtgagggt
      361 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
      421 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
      481 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
      541 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
      601 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
      661 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
      721 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
      781 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      841 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      901 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      961 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1021 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1081 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1141 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1201 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1261 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1321 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1381 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1441 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     1501 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     1561 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     1621 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     1681 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1741 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1801 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     1861 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     1921 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     1981 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     2041 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     2101 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     2161 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     2221 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     2281 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     2341 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     2401 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     2461 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     2521 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     2581 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     2641 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     2701 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     2761 catcagagca gattgtactg agagtgcacc atagatcaac gacattacta tatatataat
     2821 ataggaagca tttaatagaa cagcatcgta atatatgtgt actttgcagt tatgacgcca
     2881 gatggcagta gtggaagata ttctttattg aaaaatagct tgtcacctta cgtacaatct
     2941 tgatccggag cttttctttt tttgccgatt aagaattaat tcggtcgaaa aaagaaaagg
     3001 agagggccaa gagggagggc attggtgact attgagcacg tgagtatacg tgattaagca
     3061 cacaaaggca gcttggagta tgtctgttat taatttcaca ggtagttctg gtccattggt
     3121 gaaagtttgc ggcttgcaga gcacagaggc cgcagaatgt gctctagatt ccgatgctga
     3181 cttgctgggt attatatgtg tgcccaatag aaagagaaca attgacccgg ttattgcaag
     3241 gaaaatttca agtcttgtaa aagcatataa aaatagttca ggcactccga aatacttggt
     3301 tggcgtgttt cgtaatcaac ctaaggagga tgttttggct ctggtcaatg attacggcat
     3361 tgatatcgtc caactgcatg gagatgagtc gtggcaagaa taccaagagt tcctcggttt
     3421 gccagttatt aaaagactcg tatttccaaa agactgcaac atactactca gtgcagcttc
     3481 acagaaacct cattcgttta ttcccttgtt tgattcagaa gcaggtggga caggtgaact
     3541 tttggattgg aactcgattt ctgactgggt tggaaggcaa gagagccccg aaagcttaca
     3601 ttttatgtta gctggtggac tgacgccaga aaatgttggt gatgcgctta gattaaatgg
     3661 cgttattggt gttgatgtaa gcggaggtgt ggagacaaat ggtgtaaaag actctaacaa
     3721 aatagcaaat ttcgtcaaaa atgctaagaa ataggttatt actgagtagt atttatttaa
     3781 gtattgtttg tgcacttgcc gatctatgcg gtgtgaaata ccgcacagat gcgtaaggag
     3841 aaaataccgc atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3901 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3961 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     4021 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     4081 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     4141 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     4201 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     4261 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc
     4321 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     4381 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     4441 agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg
     4501 aattgggtac cgggcccccc ctcgaggtcg acggtatcga taagcttgat atcgaattcc
     4561 ttgaattttc aaaaattctt actttttttt tggatggacg caaagaagtt taataatcat
     4621 attacatggc attaccacca tatacatatc catatacata tccatatcta atcttactta
     4681 tatgttgtgg aaatgtaaag agccccatta tcttagccta aaaaaacctt ctctttggaa
     4741 ctttcagtaa tacgcttaac tgctcattgc tatattgaag tacggattag aagccgccga
     4801 gcgggtgaca gccctccgaa ggaagactct cctccgtgcg tcctcgtctt caccggtcgc
     4861 gttcctgaaa cgcagatgtg cctcgcgccg cactgctccg aacaataaag attctacaat
     4921 actagctttt atggttatga agaggaaaaa ttggcagtaa cctggcccca caaaccttca
     4981 aatgaacgaa tcaaattaac aaccatagga tgataatgcg attagttttt tagccttatt
     5041 tctggggtaa ttaatcagcg aagcgatgat ttttgatcta ttaacagata tataaatgca
     5101 aaaactgcat aaccacttta actaatactt tcaacatttt cggtttgtat tacttcttat
     5161 tcaaatgtaa taaaagtatc aacaaaaaat tgttaatata cctctatact ttaacgtcaa
     5221 ggagaaaaaa ccccggatcc atggactaca aagaccatga cggtgattat aaagatcatg
     5281 acatcgacta caaggatgac gatgacaagc gatccactag tcatatggat agtacgaatt
     5341 tgaacaaacg gcccttatta caatatagtc tcagttcatt gggctcgcaa ataacaaaat
     5401 ggagctcatc tagaccgact tcgccagttc gtaaggcgag aagcactgaa aatgactttc
     5461 tttccaagca agatacgtct agtatcctcc caagtatcaa cgacgacggc ggtgaacagt
     5521 ggtacgaaaa gttcaagccc aattgtttgg agcaagtggc catacataaa agaaaactta
     5581 aagatgtaca agaagcttta gatgccatgt ttttacctaa cgccaagcat aggatcctac
     5641 tactgtctgg ccccagtgga tgctctaaaa gtacggtcat aaaagaactc tcaaaaatct
     5701 tagttcctaa atacagacaa aacagcaacg gaacgtcctt tcgaagcacc ccgaacgagc
     5761 ataaagtgac cgagtttaga ggtgattgta tagtcaacga tcttcctcag atggaaagct
     5821 ttagtgagtt cttaaaaggc gcacggtatc ttgtgatgtc caacctgtca ttaatactta
     5881 tcgaggacct tcccaacgtc ttccatatag ataccagacg tcgatttcaa caacttatat
     5941 tacagtggct atatagttcg gagcctctat tacctcccct tgttatatgt ataactgaat
     6001 gtgaaattcc agagaacgat aataattatc gcaaatttgg tattgattat acatttagtg
     6061 cagaaaccat aatgaacaaa gaaatattga tgcatccaag gttgaaaaga attaagttta
     6121 atccaattaa cagcacttta ttaaaaaagc acttgaaatt tatttgtgta cagaatatga
     6181 aaatgttgaa ggagaaaaat aaatggaata aaagacagga agtcatagat tatattgcgc
     6241 aagagactgg tgatattagg tcggccatta cgacccttca attttgggcg acatcaagtg
     6301 gaagtttgcc gatttcaacc cgagaatcca ccatatcata ctttcatgcc attgggaagg
     6361 tgatacatgg ttcccatagc acgaataacg ataacgaaat gattaataac ctcttcgaaa
     6421 attcgaacaa tttgttatcg aaagaggatt tcaaattagg aatattagag aactataaca
     6481 catttaataa aggcgaattc agcatttctg atgcatcatc aattgtggat tgcctgagcg
     6541 agtgtgataa tatgaatggt ctaccagaat ccaatgagta tggtttacga gaagtgcgca
     6601 aaacctttcg taacatcagt aaacaaggcc ataatcatgg aacggtttat tttccaagag
     6661 aatggaaagt aagaaaatta caaaattcat ttaaagttca agctgaagat tggttaaatg
     6721 ttagtcttta taagtacaac gcggtacatt ctttcaggaa tataactcta gaatttggct
     6781 actacgcacc tctaattaga aagtgtcaga gttataaaaa aaaatacatt ctctattatt
     6841 tgaagaatct tccgagtggc tcctcggggc ccaaacaaac catggacaaa tttagtgata
     6901 taatgaaagt tgagaacgga atcgacgttg tggatcggat aggcgggcct atcgaagcac
     6961 tatctgtgga ggatggacta gcaccattga tggataatga tagcaataat tgtgaccatt
     7021 tagaggatca aaaaaaggaa agggacagaa ggcttcgcat gttgattgac caatatgaaa
     7081 gaaatgtgat gatggctaac gacgat
//
