LOCUS       sequence-482361-        8526 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Over-express Sc dpb11-FLAG in Yeast (integrated).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8526)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 8526)
  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 dpb11-FLAG-pRS405/GAL
FEATURES             Location/Qualifiers
     source          1..8526
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(83..151,152..943)
                     /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(301..320)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1114..1702
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1603..1622
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1856..1873
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1990..2011
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2026..2031,2032..2049,2050..2056)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2064..2080
                     /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     2069..2091
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2088..2104
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2088..2104
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2123..2143
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2125..2143
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(2192..2257)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     promoter        4568..5232
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     primer_bind     complement(4581..4604)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    4899..5016
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(5260..5276)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5261..5277)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(5301..5320)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5302..5320)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5330..5347)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5330..5346)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5339..5361)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5487..5942
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5574..5593)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5784..5805
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6243..6650
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6651..7745
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(8239..8258)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8358..8380
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8418..8436)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(8504..8526,1..82)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
       61 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
      121 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
      181 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
      241 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
      301 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
      361 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
      421 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
      481 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
      541 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
      601 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
      661 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
      721 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
      781 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
      841 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
      901 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
      961 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     1021 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     1081 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1141 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1201 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1261 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1321 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1381 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1441 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1501 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1561 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1621 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1681 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     1741 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     1801 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     1861 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     1921 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     1981 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     2041 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     2101 tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa agctggagct
     2161 ccaccgcggt ggcggccgct cagcagatct acttgtcatc gtcatccttg tagtcgatgt
     2221 catgatcttt ataatcaccg tcatggtctt tgtagtcaga atctaattcc tttgtctgat
     2281 ttcttgtctg tcgtctcata ggtttttcta aagaggcagt acgtttctta tcttgaatgg
     2341 agccataggt aacttgagta tgtgaaactt cctcagttct tgggatctct tctggattag
     2401 aaattgcagt cggctgctct gtaggcacat tttctgacat tattgtgccc caactcgtgg
     2461 ttcgtttaat ttgtttagga acgaaatttt ttggcagtga ctccaacttg atccttttcg
     2521 tatccggcat ttgtctctct gaagaaacat ccatgatact cgatgagaca ctacgctttt
     2581 ttcttccaga atcagtttct ctaattttct gcaataattc gtttttggta tctttcatag
     2641 agtgctccga taatgaggaa ttgtttcttt tcaaaagaat agttttgtcg atttttggat
     2701 cattgttttg gtactttttt tctgaactaa tgctggtata gtatggtttt tttatcttac
     2761 atttaaaatc gtccttgtgt cctcttgggc agataataca ccacttgata ttgtttaaaa
     2821 agatttcatt attttctcca gatgcagcgg acaataattt aaaaataaat gctggagtaa
     2881 ctaccggtat ggagtggaat ttcttgatat attcaatttt ccttttcatc gaatttctca
     2941 agattgaatt attttgaaaa tcttttctgt tattatcacc aggatcatca ttattgttat
     3001 taatgcaaaa ctgagtaaaa agatcgctaa attcattcga ccataacgga tgggttttcg
     3061 ggatactggg taaagctgct aagttgatta gcagaatgtc agtggattta ttgaggtatt
     3121 ctacataatt gatgcccatt ggttttagaa gatttaatac tttagtcaga tgtaacagct
     3181 ctacgccaga gaatccagtg atagtaacat tcagaaaagg ggaagaagaa aattcatgat
     3241 gcaataactt tgaagaagac gaaacttgaa actctattgt gctgaggaag ggttttgacc
     3301 ataaatctat tgggtgtagt aatttttgat aatacaagca gcgttcaatg aaaaattctg
     3361 taactatacc atcattatca tctataatct caggtaaatc attgaaggag tcaagagctt
     3421 tattacttgg aatgatgtaa taagaatgat cgtaaatgcc gctcaagtaa gacgtctcaa
     3481 ttttaccccc attttgaaca acaattttgg ttagtattga acggtgattg ccaggaaaaa
     3541 tatgatgaat gataaacgca caatttttga aaatcaaagt actattatca cttaaatctt
     3601 cttgtttgtt atttatcgat aatggtgact gccctagtac tgaaaaattt gtcttactat
     3661 gctgctgtag agacattgcc ttgtcccata ttttttcacc ctttggttta aatttattca
     3721 agagggatga tgttttcggg agcgaaggag tcagggtact ggaggattgc tgccgctgca
     3781 gagaggattg ggcatcgata ttggttggaa aggtggtgtt tattttatcc cagcaatcgc
     3841 aagagtttga tccgattgaa tcatagggca ggtctttaat atttggtaaa agataatagg
     3901 gatcgtacgg taacagtgcg cttctttttt ggcaatcgag aatccatttg aaatgtacaa
     3961 taggtatatt ttgttcaata gcagcgttaa ctcttgctcc tagtaaagta tccgtaacaa
     4021 aaatggatat ctgcccattc tgaccttgat catcgtcatc attgtgtgaa gaggtatctt
     4081 tgattacaaa gttttgatag ttataactag aacaccctaa ctttttaata ctcctaacca
     4141 atgaatcgat ggaagttatg ttagtgtcgg ttattctgcc aatgaagata ttgaagttat
     4201 gcaaatattt aaacgaaaat cgccggtaaa gtatcttcag catctcatag gtggatcctg
     4261 tcattgttgc cgtattagag tccggcaata tattttctcc agacagccag agctgataaa
     4321 tgtcatcaat ggcttgaatg tcaataaata tgatatcgaa tctatgcttc accgcaaatt
     4381 taaatttgtt tgtattggtg gtggacccaa caacaaggac attaacttgt ctggtcaaat
     4441 ctttagaaaa tatacctcct aacttaataa ttttctttga tattttttta gctaagattt
     4501 cattatttat tgcagtcgga caaaatgtta ttccttgaaa gggcttcata tgactagtgg
     4561 atccggggtt ttttctcctt gacgttaaag tatagaggta tattaacaat tttttgttga
     4621 tacttttatt acatttgaat aagaagtaat acaaaccgaa aatgttgaaa gtattagtta
     4681 aagtggttat gcagtttttg catttatata tctgttaata gatcaaaaat catcgcttcg
     4741 ctgattaatt accccagaaa taaggctaaa aaactaatcg cattatcatc ctatggttgt
     4801 taatttgatt cgttcatttg aaggtttgtg gggccaggtt actgccaatt tttcctcttc
     4861 ataaccataa aagctagtat tgtagaatct ttattgttcg gagcagtgcg gcgcgaggca
     4921 catctgcgtt tcaggaacgc gaccggtgaa gacgaggacg cacggaggag agtcttcctt
     4981 cggagggctg tcacccgctc ggcggcttct aatccgtact tcaatatagc aatgagcagt
     5041 taagcgtatt actgaaagtt ccaaagagaa ggttttttta ggctaagata atggggctct
     5101 ttacatttcc acaacatata agtaagatta gatatggata tgtatatgga tatgtatatg
     5161 gtggtaatgc catgtaatat gattattaaa cttctttgcg tccatccaaa aaaaaagtaa
     5221 gaatttttga aaattcaagg aattcgatat caagcttatc gataccgtcg acctcgaggg
     5281 ggggcccggt acccaattcg ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg
     5341 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     5401 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     5461 agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa gcgcggcggg
     5521 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     5581 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5641 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5701 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5761 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     5821 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     5881 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     5941 ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatcgacc
     6001 ggtcgaggag aacttctagt atatctacat acctaatatt attgccttat taaaaatgga
     6061 atcccaacaa ttacatcaaa atccacattc tcttcaaaat caattgtcct gtacttcctt
     6121 gttcatgtgt gttcaaaaac gttatattta taggataatt atactctatt tctcaacaag
     6181 taattggttg tttggccgag cggtctaagg cgcctgattc aagaaatatc ttgaccgcag
     6241 ttaactgtgg gaatactcag gtatcgtaag atgcaagagt tcgaatctct tagcaaccat
     6301 tatttttttc ctcaacataa cgagaacaca caggggcgct atcgcacaga atcaaattcg
     6361 atgactggaa attttttgtt aatttcagag gtcgcctgac gcatatacct ttttcaactg
     6421 aaaaattggg agaaaaagga aaggtgagag cgccggaacc ggcttttcat atagaataga
     6481 gaagcgttca tgactaaatg cttgcatcac aatacttgaa gttgacaata ttatttaagg
     6541 acctattgtt ttttccaata ggtggttagc aatcgtctta ctttctaact tttcttacct
     6601 tttacatttc agcaatatat atatatatat ttcaaggata taccattcta atgtctgccc
     6661 ctaagaagat cgtcgttttg ccaggtgacc acgttggtca agaaatcaca gccgaagcca
     6721 ttaaggttct taaagctatt tctgatgttc gttccaatgt caagttcgat ttcgaaaatc
     6781 atttaattgg tggtgctgct atcgatgcta caggtgttcc acttccagat gaggcgctgg
     6841 aagcctccaa gaaggctgat gccgttttgt taggtgctgt gggtggtcct aaatggggta
     6901 ccggtagtgt tagacctgaa caaggtttac taaaaatccg taaagaactt caattgtacg
     6961 ccaacttaag accatgtaac tttgcatccg actctctttt agacttatct ccaatcaagc
     7021 cacaatttgc taaaggtact gacttcgttg ttgtcagaga attagtggga ggtatttact
     7081 ttggtaagag aaaggaagac gatggtgatg gtgtcgcttg ggatagtgaa caatacaccg
     7141 ttccagaagt gcaaagaatc acaagaatgg ccgctttcat ggccctacaa catgagccac
     7201 cattgcctat ttggtccttg gataaagcta atgttttggc ctcttcaaga ttatggagaa
     7261 aaactgtgga ggaaaccatc aagaacgaat tccctacatt gaaggttcaa catcaattga
     7321 ttgattctgc cgccatgatc ctagttaaga acccaaccca cctaaatggt attataatca
     7381 ccagcaacat gtttggtgat atcatctccg atgaagcctc cgttatccca ggttccttgg
     7441 gtttgttgcc atctgcgtcc ttggcctctt tgccagacaa gaacaccgca tttggtttgt
     7501 acgaaccatg ccacggttct gctccagatt tgccaaagaa taaggtcaac cctatcgcca
     7561 ctatcttgtc tgctgcaatg atgttgaaat tgtcattgaa cttgcctgaa gaaggtaagg
     7621 ccattgaaga tgcagttaaa aaggttttgg atgcaggtat cagaactggt gatttaggtg
     7681 gttccaacag taccaccgaa gtcggtgatg ctgtcgccga agaagttaag aaaatccttg
     7741 cttaaaaaga ttctcttttt ttatgatatt tgtacataaa ctttataaat gaaattcata
     7801 atagaaacga cacgaaatta caaaatggaa tatgttcata gggtagacga aactatatac
     7861 gcaatctaca tacatttatc aagaaggaga aaaaggagga tgtaaaggaa tacaggtaag
     7921 caaattgata ctaatggctc aacgtgataa ggaaaaagaa ttgcacttta acattaatat
     7981 tgacaaggag gagggcacca cacaaaaagt taggtgtaac agaaaatcat gaaactatga
     8041 ttcctaattt atatattgga ggattttctc taaaaaaaaa aaaatacaac aaataaaaaa
     8101 cactcaatga cctgaccatt tgatggagtt taagtcaata ccttcttgaa ccatttccca
     8161 taatggtgaa agttccctca agaattttac tctgtcagaa acggccttaa cgacgtagtc
     8221 gatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     8281 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     8341 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     8401 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     8461 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     8521 tttatt
//
