LOCUS       sequence-482082-        8239 bp    DNA     circular SYN 05-AUG-2025
DEFINITION  Over-express Sc FLAG-rad30 (Sc Pol eta) in yeast (integrated).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8239)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 8239)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 5, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Sc FLAG-rad30-pRS403/GAL
FEATURES             Location/Qualifiers
     source          1..8239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1396..1461)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        1474..2138
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(1487..1510)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    1805..1922
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(2166..2182)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2167..2183)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2207..2226)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2208..2226)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2236..2253)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2236..2252)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2245..2267)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2393..2848
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2480..2499)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2690..2711
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3108..3770)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(3771..3958)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(4105..4124)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4224..4246
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4284..4302)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4370..4474
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4475..4543,4544..5335)
                     /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     complement(4693..4712)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5506..6094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5995..6014
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6248..6265
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6382..6403
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6418..6423,6424..6441,6442..6448)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6456..6472
                     /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)."
     primer_bind     6461..6483
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6480..6496
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6480..6496
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     6515..6535
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        6517..6535
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 tggacctgat tttcgatata cttcgtagga ttttgttttt aaagatatag atactgttct
       61 tggtattacg atcttgttat actcctgttc cagatcctga atcctggacg taagctcagc
      121 gcagaagacc tcaagccaag aaatacagtc cacgatggaa ttgcacgatt ttcctctaag
      181 atttttgttt gacatcatac tttttacaac agggcgagaa cttaagggca gaccgtaccg
      241 gcctctactt aatttaaaca acttttctgc taaatctgct gttttcaaag ggtcgatatt
      301 gctggttgag cgatcataat cagattgctt aactttagca tctaaaaatt ccttaagctg
      361 gccagcatta tcaggccacg tctctctgat gtgtttaata ctgttttcgt ggggaagatc
      421 caatacgtca attagttctt tgccaagaac gccgcctaag gtccagaatg atgtaatttc
      481 aaactttccg caatctaaaa aatctaataa acaatcattt ttcacaattg tttgggcatc
      541 aggttttttg taattagaag ccaacttgca aacatttttg gtgctagaca aaccacacga
      601 agtagtataa ccgagaatat cttttatgct atctctgata cccttgcata cctgagatcc
      661 tagtgcaagt attacatcgt cccaatctgt gatcagatct ctgccctctg gattgaaaac
      721 atcgccttca aacttcagag actttatctt ttcgggtata agaggtagat gggaatttat
      781 atcatagttg ccccctataa aagcctcacg aatattgctt aatgcatctt tcagtttcaa
      841 gtcgcctgta agctcgtact cattatcaaa cattagcata ttgaaacaaa tccggcccaa
      901 atcaagaaat acttcatcaa tactcgccct ttcgactaga tcacatgcgc ttttgaaaat
      961 cttcagcgcc ttgcgacttt ccctcctata tggttccagt gatactttat gatcttccac
     1021 acttatctgc tttgcaggat cctgcaccca agatccacat ccatcatgat attgccaaaa
     1081 atcttctccc tttttaaaaa cggctgtatg gatcggtatg aggttactac atttttttag
     1141 tgcttcctga atcgtatcca tccttgatat accatacttt cttgctgcat aagagactgc
     1201 aataatggaa ttccactgaa cgcagactac cggatcttct ttgcttaagc cacatcgcat
     1261 ctgctcaacc tgtgcaaaaa aggcattcat atctatgtgg gcgatacatg ctaaggagga
     1321 ctcgtatgct ttactggggg aaccaagctg aatcaactcc ttccaagtaa attttgacat
     1381 atgactagtg gatcgcttgt catcgtcatc cttgtagtcg atgtcatgat ctttataatc
     1441 accgtcatgg tctttgtagt ccatggatcc ggggtttttt ctccttgacg ttaaagtata
     1501 gaggtatatt aacaattttt tgttgatact tttattacat ttgaataaga agtaatacaa
     1561 accgaaaatg ttgaaagtat tagttaaagt ggttatgcag tttttgcatt tatatatctg
     1621 ttaatagatc aaaaatcatc gcttcgctga ttaattaccc cagaaataag gctaaaaaac
     1681 taatcgcatt atcatcctat ggttgttaat ttgattcgtt catttgaagg tttgtggggc
     1741 caggttactg ccaatttttc ctcttcataa ccataaaagc tagtattgta gaatctttat
     1801 tgttcggagc agtgcggcgc gaggcacatc tgcgtttcag gaacgcgacc ggtgaagacg
     1861 aggacgcacg gaggagagtc ttccttcgga gggctgtcac ccgctcggcg gcttctaatc
     1921 cgtacttcaa tatagcaatg agcagttaag cgtattactg aaagttccaa agagaaggtt
     1981 tttttaggct aagataatgg ggctctttac atttccacaa catataagta agattagata
     2041 tggatatgta tatggatatg tatatggtgg taatgccatg taatatgatt attaaacttc
     2101 tttgcgtcca tccaaaaaaa aagtaagaat ttttgaaaat tcaaggaatt cgatatcaag
     2161 cttatcgata ccgtcgacct cgaggggggg cccggtaccc aattcgccct atagtgagtc
     2221 gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
     2281 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
     2341 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc
     2401 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2461 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2521 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2581 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2641 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2701 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2761 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2821 tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt
     2881 gcggtatttc acaccgcata gatccgtcga gttcaagaga aaaaaaaaga aaaagcaaaa
     2941 agaaaaaagg aaagcgcgcc tcgttcagaa tgacacgtat agaatgatgc attaccttgt
     3001 catcttcagt atcatactgt tcgtatacat acttactgac attcataggt atacatatat
     3061 acacatgtat atatatcgta tgctgcagct ttaaataatc ggtgtcacta cataagaaca
     3121 cctttggtgg agggaacatc gttggtacca ttgggcgagg tggcttctct tatggcaacc
     3181 gcaagagcct tgaacgcact ctcactacgg tgatgatcat tcttgcctcg cagacaatca
     3241 acgtggaggg taattctgct agcctctgca aagctttcaa gaaaatgcgg gatcatctcg
     3301 caagagagat ctcctacttt ctccctttgc aaaccaagtt cgacaactgc gtacggcctg
     3361 ttcgaaagat ctaccaccgc tctggaaagt gcctcatcca aaggcgcaaa tcctgatcca
     3421 aaccttttta ctccacgcac ggcccctagg gcctctttaa aagcttgacc gagagcaatc
     3481 ccgcagtctt cagtggtgtg atggtcgtct atgtgtaagt caccaatgca ctcaacgatt
     3541 agcgaccagc cggaatgctt ggccagagca tgtatcatat ggtccagaaa ccctatacct
     3601 gtgtggacgt taatcacttg cgattgtgtg gcctgttctg ctactgcttc tgcctctttt
     3661 tctgggaaga tcgagtgctc tatcgctagg ggaccaccct ttaaagagat cgcaatctga
     3721 atcttggttt catttgtaat acgctttact agggctttct gctctgtcat ctttgccttc
     3781 gtttatcttg cctgctcatt ttttagtata ttcttcgaag aaatcacatt actttatata
     3841 atgtataatt cattatgtga taatgccaat cgctaagaaa aaaaaagagt catccgctag
     3901 gtggaaaaaa aaaaatgaaa atcattaccg aggcataaaa aaatatagag tgtactagag
     3961 gaggccaaga gtaatagaaa aagaaaattg cgggaaagga ctgtgttatg acttccctga
     4021 ctaatgccgt gttcaaacga tacctggcag tgactcctag cgctcaccaa gctcttaaaa
     4081 cgggaattta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4141 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4201 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4261 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4321 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4381 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4441 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4501 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4561 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4621 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4681 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4741 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4801 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4861 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4921 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4981 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5041 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5101 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5161 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5221 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5281 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5341 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5401 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5461 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5521 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5581 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5641 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5701 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5761 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5821 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5881 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5941 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6001 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6061 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6121 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6181 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     6241 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     6301 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     6361 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     6421 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     6481 aggaaacagc tatgaccatg attacgccaa gcgcgcaatt aaccctcact aaagggaaca
     6541 aaagctggag ctccaccgcg gtggcggccg ctcagcagat ccttataaca atccactcaa
     6601 ggtcagcttc tccaggaatg gtgaaagatc ttcacgacgt atttcaatta tggaactttc
     6661 aaaattatct attctcactt tagcatccgt ttctgcagtt atttgttgaa acgccatgct
     6721 gaaattattc aaaataggtt tttttgtgtg tatcgtgtct agtccgcgag gatacctagg
     6781 tgttaaccca gcaaccaagt aagtgtccgt attttcatct atgctggcaa gcaccatggg
     6841 caacaattgc ttgtcttcag attccgcaca gcattctatg agccaatttc ctaatcttaa
     6901 taacgtcaat gggtttctgt acaagtctaa atcgggtccg tcttgtaaga cgcataatct
     6961 ataaattctt aaatgcttga ttaatttctt ctccaatata gcaaccccgg tattaaaaat
     7021 tgctctctgt aagtcttgtg ctagttgaat gccacggttt aatagttcca cctttctgtc
     7081 atctagagca tcccaactga gccaaaaatt cgaaacccat ctttttctca aattcgtcaa
     7141 tttttgagcg ctgttatctt cttcttcctc tccatctgta tcgtcattat tgtcattatt
     7201 tattttcaca ctatccttat cagttgaatt acctacttcc aaaagagctg ttagtgcttc
     7261 gacaaattca ctggcactta tggacccacg atatccaaga gttctaacaa acccatctct
     7321 aattatatct tgcaacccat aacgatccaa atttttgtca aatattatcc caagttctct
     7381 cttaatagaa tgatccatgt ataaccatgt ttcttgtgca gtacttaatg gtatacccat
     7441 tctagcaaac atcttatgca atctcttttt cccattttca ttccataggg atagtttggc
     7501 attgacatag ttagaataat aaaaactgtc gtacaatgaa gaatgcctca gtaagaaaag
     7561 gtagtaatcc ggttggatgt ttaacgtcaa tgtatcaggg gtctttactg agtttctact
     7621 gcttggtgtg agacgcttca cttcatcttg caataagggg tataatcgat tgtaaacttg
     7681 agcatatgct gaagccatat aattgtctat ttggaatagg tcattttttt cttgtaaaaa
     7741 atgataagat gtttttggaa gatgtaactt gtttcttctg aggtgtggca gtatgttgtg
     7801 agtttggtct tttccgtgaa aacaacaatc ttttttcccc gaaagaaagg tttttggatg
     7861 attcttcagc accatttaag ccttccgaca gtttcaatgc taaatgatag tctgcgtgct
     7921 cttgaagagc cttctggtct gtaaaagtca cctggtattt acaacattcc aacttcggag
     7981 ttttctcgtc agctttcgag ctggtagttt tttcctcatc ttccttgcct gctgagcttt
     8041 tgaaagtatg gacctgattg ccaaacatat ctacaacagt tttttgtaaa tctataatat
     8101 cgaaattagt aattgtcata cttagttttg ttaaagggta ataacttttg ttctttcctt
     8161 taatatcaag gtccgtgaca aattttatgc ctacttttag gagttcgtga ctctggaaat
     8221 tgatgccctt gtacgcaac
//
