pGAD-T7-A-bkBE

LOCUS     pGAD-T7-A-bkBE              8001 bp DNA     circular SYN 19-MAR-2026
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8001)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Oct 1, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        771..1475
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1521..1541
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1557..1898
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     promoter        1904..1922
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1929..1938
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1941..1967
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1989..2036
                     /label=AD-BK
     terminator      2429..2616
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(2733..3827)
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIEATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     promoter        complement(3828..4233)
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     primer_bind     complement(4276..4291)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4299..4315
                     /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(4323..4353)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4368..4389
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     protein_bind    complement(4444..4477)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(4555..4588)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     rep_origin      complement(4828..5416)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5587..6447)
                     /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"
     promoter        complement(6448..6552)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6834..7998
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tgcatgcctg caggtcgaga tccgggatcg aagaaatgat ggtaaatgaa ataggaaatc
       61 aaggagcatg aaggcaaaag acgaatataa gggtcgaacg aaaaataaag tgaaaagtgt
      121 tgatatgatg tatttggctt tgcggcgccg aaaaaacgag tttacgcaat tgcacaatca
      181 tgctgactct gtggcggacc cgcgctcttg ccggcccggc gataacgctg ggcgtgaggc
      241 tgtgcccggc ggagtttttt gcgcctgcat tttccaaggt ttaccctgcg ctaaggggcg
      301 agattggaga agcaataaga atgccggttg gggttgcgat gatgacgacc acgacaactg
      361 gtgtcattat ttaagttgcc gaaagaacct gagtgcattt gcaacatgag tatactagaa
      421 gaatgagcca agacttgcga gacgcgagtt tgccggtggt gcgaacaata gagcgaccat
      481 gaccttgaag gtgagacgcg cataaccgct agagtacttt gaagaggaaa cagcaatagg
      541 gttgctacca gtataaatag acaggtacat acaacactgg aaatggttgt ctgtttgagt
      601 acgctttcaa ttcatttggg tgtgcacttt attatgttac aatatggaag ggaactttac
      661 acttctccta tgcacatata ttaattaaag tccaatgcta gtagagaagg ggggtaacac
      721 ccctccgcgc tcttttccga tttttttcta aaccgtggaa tatttcggat atccttttgt
      781 tgtttccggg tgtacaatat ggacttcctc ttttctggca accaaaccca tacatcggga
      841 ttcctataat accttcgttg gtctccctaa catgtaggtg gcggagggga gatatacaat
      901 agaacagata ccagacaaga cataatgggc taaacaagac tacaccaatt acactgcctc
      961 attgatggtg gtacataacg aactaatact gtagccctag acttgatagc catcatcata
     1021 tcgaagtttc actacccttt ttccatttgc catctattga agtaataata ggcgcatgca
     1081 acttcttttc tttttttttc ttttctctct cccccgttgt tgtctcacca tatccgcaat
     1141 gacaaaaaaa tgatggaaga cactaaagga aaaaattaac gacaaagaca gcaccaacag
     1201 atgtcgttgt tccagagctg atgaggggta tctcgaagca cacgaaactt tttccttcct
     1261 tcattcacgc acactactct ctaatgagca acggtatacg gccttccttc cagttacttg
     1321 aatttgaaat aaaaaaaagt ttgctgtctt gctatcaagt ataaatagac ctgcaattat
     1381 taatcttttg tttcctcgtc attgttctcg ttccctttct tccttgtttc tttttctgca
     1441 caatatttca agctatacca agcatacaat caactccaag ctttgcaaag atggataaag
     1501 cggaattaat tcccgagcct ccaaaaaaga agagaaaggt cgaattgggt accgccgcca
     1561 attttaatca aagtgggaat attgctgata gctcattgtc cttcactttc actaacagta
     1621 gcaacggtcc gaacctcata acaactcaaa caaattctca agcgctttca caaccaattg
     1681 cctcctctaa cgttcatgat aacttcatga ataatgaaat cacggctagt aaaattgatg
     1741 atggtaataa ttcaaaacca ctgtcacctg gttggacgga ccaaactgcg tataacgcgt
     1801 ttggaatcac tacagggatg tttaatacca ctacaatgga tgatgtatat aactatctat
     1861 tcgatgatga agatacccca ccaaacccaa aaaaagagat ctttaatacg actcactata
     1921 gggcgagcgc cgccatggag tacccatacg acgtaccaga ttacgctcat atggccatgg
     1981 aggccagtca ggtcgactct cgaggatccg aattcggtac ccggtatcga tttaaaatcg
     2041 agctcgagct gcagatgaat cgtagatact gaaaaacccc gcaagttcac ttcaactgtg
     2101 catcgtgcac catctcaatt tctttcattt atacatcgtt ttgccttctt ttatgtaact
     2161 atactcctct aagtttcaat cttggccatg taacctctga tctatagaat tttttaaatg
     2221 actagaatta atgcccatct tttttttgga cctaaattct tcatgaaaat atattacgag
     2281 ggcttattca gaagctttgg acttcttcgc cagaggtttg gtcaagtctc caatcaaggt
     2341 tgtcggcttg tctaccttgc cagaaattta cgaaaagatg gaaaagggtc aaatcgttgg
     2401 tagatacgtt gttgacactt ctaaataagc gaatttctta tgatttatga tttttattat
     2461 taaataagtt ataaaaaaaa taagtgtata caaattttaa agtgactctt aggttttaaa
     2521 acgaaaattc ttattcttga gtaactcttt cctgtaggtc aggttgcttt ctcaggtata
     2581 gcatgaggtc gctcttattg accacacctc taccggccgg tcgaaattcc cctaccctat
     2641 gaacatattc cattttgtaa tttcgtgtcg tttctattat gaatttcatt tataaagttt
     2701 atgtacaaat atcataaaaa aagagaatct ttttaagcaa ggattttctt aacttcttcg
     2761 gcgacagcat caccgacttc ggtggtactg ttggaaccac ctaaatcacc agttctgata
     2821 cctgcatcca aaaccttttt aactgcatct tcaatggcct taccttcttc aggcaagttc
     2881 aatgacaatt tcaacatcat tgcagcagac aagatagtgg cgatagggtt gaccttattc
     2941 tttggcaaat ctggagcaga accgtggcat ggttcgtaca aaccaaatgc ggtgttcttg
     3001 tctggcaaag aggccaagga cgcagatggc aacaaaccca aggaacctgg gataacggag
     3061 gcttcatcgg agatgatatc accaaacatg ttgctggtga ttataatacc atttaggtgg
     3121 gttgggttct taactaggat catggcggca gaatcaatca attgatgttg aaccttcaat
     3181 gtaggaaatt cgttcttgat ggtttcctcc acagtttttc tccataatct tgaagaggcc
     3241 aaaacattag ctttatccaa ggaccaaata ggcaatggtg gctcatgttg tagggccatg
     3301 aaagcggcca ttcttgtgat tctttgcact tctggaacgg tgtattgttc actatcccaa
     3361 gcgacaccat caccatcgtc ttcctttctc ttaccaaagt aaatacctcc cactaattct
     3421 ctgacaacaa cgaagtcagt acctttagca aattgtggct tgattggaga taagtctaaa
     3481 agagagtcgg atgcaaagtt acatggtctt aagttggcgt acaattgaag ttctttacgg
     3541 atttttagta aaccttgttc aggtctaaca ctacctgtac cccatttagg accacccaca
     3601 gcacctaaca aaacggcatc agccttcttg gaggcttcca gcgcctcatc tggaagtggg
     3661 acacctgtag cttcgatagc agcaccacca attaaatgat tttcgaaatc gaacttgaca
     3721 ttggaacgaa catcagaaat agctttaaga accttaatgg cttcggctgt gatttcttga
     3781 ccaacgtggt cacctggcaa aacgacgatc ttcttagggg cagacattag aatggtatat
     3841 ccttgaaata tatatatata ttgctgaaat gtaaaaggta agaaaagtta gaaagtaaga
     3901 cgattgctaa ccacctattg gaaaaaacaa taggtcctta aataatattg tcaacttcaa
     3961 gtattgtgat gcaagcattt agtcatgaac gcttctctat tctatatgaa aagccggttc
     4021 cggcgctctc acctttcctt tttctcccaa tttttcagtt gaaaaaggta tatgcgtcag
     4081 gcgacctctg aaattaacaa aaaatttcca gtcatcgaat ttgattctgt gcgatagcgc
     4141 ccctgtgtgt tctcgttatg ttgaggaaaa aaataatggt tgctaagaga ttcgaactct
     4201 tgcatcttac gatacctgag tattcccaca gttgggggat ctcgactcta gctagaggat
     4261 caattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4321 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4381 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4441 ctgataactt cgtataatgt atgctatacg aagttattag gtctgaagag gagtttacgt
     4501 ccagccaagc tagcttggct gcaggtcgag cggccgcgat ccggaaccct taatataact
     4561 tcgtataatg tatgctatac gaagttatca gctgcattaa tgaatcggcc aacgcgcggg
     4621 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4681 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4741 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4801 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4861 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4921 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4981 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     5041 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5101 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5161 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5221 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     5281 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5341 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5401 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5461 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5521 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5581 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5641 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5701 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     5761 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5821 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5881 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5941 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6001 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6061 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6121 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6181 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6241 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6301 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6361 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6421 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6481 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6541 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     6601 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     6661 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     6721 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     6781 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc ataacgcatt
     6841 taagcataaa cacgcactat gccgttcttc tcatgtatat atatatacag gcaacacgca
     6901 gatataggtg cgacgtgaac agtgagctgt atgtgcgcag ctcgcgttgc attttcggaa
     6961 gcgctcgttt tcggaaacgc tttgaagttc ctattccgaa gttcctattc tctagctaga
     7021 aagtatagga acttcagagc gcttttgaaa accaaaagcg ctctgaagac gcactttcaa
     7081 aaaaccaaaa acgcaccgga ctgtaacgag ctactaaaat attgcgaata ccgcttccac
     7141 aaacattgct caaaagtatc tctttgctat atatctctgt gctatatccc tatataacct
     7201 acccatccac ctttcgctcc ttgaacttgc atctaaactc gacctctaca ttttttatgt
     7261 ttatctctag tattactctt tagacaaaaa aattgtagta agaactattc atagagtgaa
     7321 tcgaaaacaa tacgaaaatg taaacatttc ctatacgtag tatatagaga caaaatagaa
     7381 gaaaccgttc ataattttct gaccaatgaa gaatcatcaa cgctatcact ttctgttcac
     7441 aaagtatgcg caatccacat cggtatagaa tataatcggg gatgccttta tcttgaaaaa
     7501 atgcacccgc agcttcgcta gtaatcagta aacgcgggaa gtggagtcag gcttttttta
     7561 tggaagagaa aatagacacc aaagtagcct tcttctaacc ttaacggacc tacagtgcaa
     7621 aaagttatca agagactgca ttatagagcg cacaaaggag aaaaaaagta atctaagatg
     7681 ctttgttaga aaaatagcgc tctcgggatg catttttgta gaacaaaaaa gaagtataga
     7741 ttctttgttg gtaaaatagc gctctcgcgt tgcatttctg ttctgtaaaa atgcagctca
     7801 gattctttgt ttgaaaaatt agcgctctcg cgttgcattt ttgttttaca aaaatgaagc
     7861 acagattctt cgttggtaaa atagcgcttt cgcgttgcat ttctgttctg taaaaatgca
     7921 gctcagattc tttgtttgaa aaattagcgc tctcgcgttg catttttgtt ctacaaaatg
     7981 aagcacagat gcttcgttgc t
//