LOCUS       sequence-482352-        7148 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Over-express FLAG-S6-rad24 in yeast (integrated).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7148)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 7148)
  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-S6-rad24- pRS404/GAL-L
FEATURES             Location/Qualifiers
     source          1..7148
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(49..840,841..909))
                     /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     672..691
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(910..1014)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1082..1100
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1138..1160)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1260..1279
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1300..1581
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             1582..2256
                     /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(2359..2814)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2496..2517)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2708..2727
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2940..2962
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2954..2971
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2955..2971
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2981..3000
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2981..2999
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3024..3040
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     3025..3041
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3069..3733
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    3285..3402
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3697..3720
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             3746..3811
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(5997..6017)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5997..6015)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6036..6052)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6036..6052)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6049..6071)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6060..6076
                     /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(6084..6090,6091..6108,6109..6114))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6129..6150
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6267..6284)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6438..7026)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6518..6537)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
       61 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      121 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      181 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      241 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      301 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      361 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      421 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      481 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
      541 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
      601 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
      661 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
      721 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
      781 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
      841 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
      901 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
      961 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1021 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     1081 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
     1141 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
     1201 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
     1261 ggcatcagag cagattgtac tgagagtgca ccatagatca acgacattac tatatatata
     1321 atataggaag catttaatag aacagcatcg taatatatgt gtactttgca gttatgacgc
     1381 cagatggcag tagtggaaga tattctttat tgaaaaatag cttgtcacct tacgtacaat
     1441 cttgatccgg agcttttctt tttttgccga ttaagaatta attcggtcga aaaaagaaaa
     1501 ggagagggcc aagagggagg gcattggtga ctattgagca cgtgagtata cgtgattaag
     1561 cacacaaagg cagcttggag tatgtctgtt attaatttca caggtagttc tggtccattg
     1621 gtgaaagttt gcggcttgca gagcacagag gccgcagaat gtgctctaga ttccgatgct
     1681 gacttgctgg gtattatatg tgtgcccaat agaaagagaa caattgaccc ggttattgca
     1741 aggaaaattt caagtcttgt aaaagcatat aaaaatagtt caggcactcc gaaatacttg
     1801 gttggcgtgt ttcgtaatca acctaaggag gatgttttgg ctctggtcaa tgattacggc
     1861 attgatatcg tccaactgca tggagatgag tcgtggcaag aataccaaga gttcctcggt
     1921 ttgccagtta ttaaaagact cgtatttcca aaagactgca acatactact cagtgcagct
     1981 tcacagaaac ctcattcgtt tattcccttg tttgattcag aagcaggtgg gacaggtgaa
     2041 cttttggatt ggaactcgat ttctgactgg gttggaaggc aagagagccc cgaaagctta
     2101 cattttatgt tagctggtgg actgacgcca gaaaatgttg gtgatgcgct tagattaaat
     2161 ggcgttattg gtgttgatgt aagcggaggt gtggagacaa atggtgtaaa agactctaac
     2221 aaaatagcaa atttcgtcaa aaatgctaag aaataggtta ttactgagta gtatttattt
     2281 aagtattgtt tgtgcacttg ccgatctatg cggtgtgaaa taccgcacag atgcgtaagg
     2341 agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2401 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2461 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2521 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2581 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2641 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2701 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2761 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc
     2821 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     2881 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     2941 ccagtcacga cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg
     3001 cgaattgggt accgggcccc ccctcgaggt cgacggtatc gataagcttg atatcgaatt
     3061 ccttgaattt tcaaaaattc ttactttttt tttggatgga cgcaaagaag tttaataatc
     3121 atattacatg gcattaccac catatacata tccatataca tatccatatc taatcttact
     3181 tatatgttgt ggaaatgtaa agagccccat tatcttagcc taaaaaaacc ttctctttgg
     3241 aactttcagt aatacgctta actgctcatt gctatattga agtacggatt agaagccgcc
     3301 gagcgggtga cagccctccg aaggaagact ctcctccgtg cgtcctcgtc ttcaccggtc
     3361 gcgttcctga aacgcagatg tgcctcgcgc cgcactgctc cgaacaataa agattctaca
     3421 atactagctt ttatggttat gaagaggaaa aattggcagt aacctggccc cacaaacctt
     3481 caaatgaacg aatcaaatta acaaccatag gatgataatg cgattagttt tttagcctta
     3541 tttctggggt aattaatcag cgaagcgatg atttttgatc tattaacaga tatataaatg
     3601 caaaaactgc ataaccactt taactaatac tttcaacatt ttcggtttgt attacttctt
     3661 attcaaatgt aataaaagta tcaacaaaaa attgttaata tacctctata ctttaacgtc
     3721 aaggagaaaa aaccccggat ccatggacta caaagaccat gacggtgatt ataaagatca
     3781 tgacatcgac tacaaggatg acgatgacaa gcgatccact agtcatatcg gcgatagcct
     3841 gagctggctg ctgcgcctgc tgaaccatat ggatagtacg aatttgaaca aacggccctt
     3901 attacaatat agtctcagtt cattgggctc gcaaataaca aaatggagct catctagacc
     3961 gacttcgcca gttcgtaagg cgagaagcac tgaaaatgac tttctttcca agcaagatac
     4021 gtctagtatc ctcccaagta tcaacgacga cggcggtgaa cagtggtacg aaaagttcaa
     4081 gcccaattgt ttggagcaag tggccataca taaaagaaaa cttaaagatg tacaagaagc
     4141 tttagatgcc atgtttttac ctaacgccaa gcataggatc ctactactgt ctggccccag
     4201 tggatgctct aaaagtacgg tcataaaaga actctcaaaa atcttagttc ctaaatacag
     4261 acaaaacagc aacggaacgt cctttcgaag caccccgaac gagcataaag tgaccgagtt
     4321 tagaggtgat tgtatagtca acgatcttcc tcagatggaa agctttagtg agttcttaaa
     4381 aggcgcacgg tatcttgtga tgtccaacct gtcattaata cttatcgagg accttcccaa
     4441 cgtcttccat atagatacca gacgtcgatt tcaacaactt atattacagt ggctatatag
     4501 ttcggagcct ctattacctc cccttgttat atgtataact gaatgtgaaa ttccagagaa
     4561 cgataataat tatcgcaaat ttggtattga ttatacattt agtgcagaaa ccataatgaa
     4621 caaagaaata ttgatgcatc caaggttgaa aagaattaag tttaatccaa ttaacagcac
     4681 tttattaaaa aagcacttga aatttatttg tgtacagaat atgaaaatgt tgaaggagaa
     4741 aaataaatgg aataaaagac aggaagtcat agattatatt gcgcaagaga ctggtgatat
     4801 taggtcggcc attacgaccc ttcaattttg ggcgacatca agtggaagtt tgccgatttc
     4861 aacccgagaa tccaccatat catactttca tgccattggg aaggtgatac atggttccca
     4921 tagcacgaat aacgataacg aaatgattaa taacctcttc gaaaattcga acaatttgtt
     4981 atcgaaagag gatttcaaat taggaatatt agagaactat aacacattta ataaaggcga
     5041 attcagcatt tctgatgcat catcaattgt ggattgcctg agcgagtgtg ataatatgaa
     5101 tggtctacca gaatccaatg agtatggttt acgagaagtg cgcaaaacct ttcgtaacat
     5161 cagtaaacaa ggccataatc atggaacggt ttattttcca agagaatgga aagtaagaaa
     5221 attacaaaat tcatttaaag ttcaagctga agattggtta aatgttagtc tttataagta
     5281 caacgcggta cattctttca ggaatataac tctagaattt ggctactacg cacctctaat
     5341 tagaaagtgt cagagttata aaaaaaaata cattctctat tatttgaaga atcttccgag
     5401 tggctcctcg gggcccaaac aaaccatgga caaatttagt gatataatga aagttgagaa
     5461 cggaatcgac gttgtggatc ggataggcgg gcctatcgaa gcactatctg tggaggatgg
     5521 actagcacca ttgatggata atgatagcaa taattgtgac catttagagg atcaaaaaaa
     5581 ggaaagggac agaaggcttc gcatgttgat tgaccaatat gaaagaaatg tgatgatggc
     5641 taacgacgat cttgaagacg aagaaacttc ttttaatgat gaccctattg tcgatagcga
     5701 tagcgataac agcaataata ttggcaatga aacatttggt agaagcgacg aagacgagtc
     5761 tctatgtgaa attctgtccc agagacagcc gcgtaaagcg ccagttatca gtgagtccct
     5821 ttcagattca gatctggaaa tactctaact ttttactctt taaatttgac gagaaaaccc
     5881 caggaaatat ccacacaaat ctatgatccg gctgctaaca aagcccgaaa ggaagctgag
     5941 ttggctgctg ccaccgctga gcggccgcca ccgcggtgga gctccagctt ttgttccctt
     6001 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6061 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6121 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6181 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6241 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6301 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6361 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6421 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6481 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     6541 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     6601 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     6661 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     6721 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     6781 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     6841 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     6901 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     6961 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7021 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7081 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7141 taaaaatg
//
