LOCUS       pGADT7-Rec.        8058 bp DNA     circular SYN 01-JAN-1980
DEFINITION  High-copy yeast vector for fusing a cDNA to the GAL4 activation 
            domain by in vivo recombination.
ACCESSION   .
VERSION     .
KEYWORDS    pGADT7-Rec.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8058)
  AUTHORS   Clontech
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8058)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Linearize with SmaI.
FEATURES             Location/Qualifiers
     source          1..8058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        771..1475
                     /label=ADH1 promoter
                     /note="promoter for alcohol dehydrogenase 1"
     CDS             1491..1493
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             1521..1541
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             1557..1898
                     /label=GAL4 activation domain
                     /note="activation domain of the GAL4 transcriptional
                     activator"
     promoter        1904..1922
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1935..1937
                     /codon_start=1
                     /product="in vitro start codon"
                     /label=in vitro start codon
                     /note="ATG"
                     /translation="M"
     CDS             1941..1967
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
     misc_feature    2000..2035
                     /label=SMART III Oligonucleotide Sequence
                     /note="SMART III"
                     /note="matches the SMART III Oligonucleotide for cDNA
                     synthesis"
     misc_feature    2046..2070
                     /label=CDS III Sequence
                     /note="CDS III"
                     /note="matches the CDS III Primer for cDNA synthesis"
     terminator      2486..2673
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae
                     alcohol dehydrogenase 1 (ADH1) gene"
     CDS             complement(2793..3884)
                     /label=LEU2
                     /note="3-isopropylmalate dehydrogenase, required for
                     leucine biosynthesis"
     promoter        complement(3885..4290)
                     /label=LEU2 promoter
     primer_bind     complement(4332..4348)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4356..4372)
                     /label=lac operator
                     /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(4380..4410)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4425..4446)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     protein_bind    4501..4534
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     rep_origin      complement(4885..5473)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5647..6504)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6505..6609)
                     /label=AmpR promoter
     rep_origin      6891..8055
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
ORIGIN
        1 tgcatgcctg caggtcgaga tccgggatcg aagaaatgat ggtaaatgaa ataggaaatc
       61 aaggagcatg aaggcaaaag acaaatataa 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 aggccagtga attccaccca agcagtggta tcaacgcaga gtggccatta tggcccggga
     2041 aaaaacatgt cggccgcctc ggcctctaga gggtgggcat cgatacggga tccatcgagc
     2101 tcgagctgca gatgaatcgt agatactgaa aaaccccgca agttcacttc aactgtgcat
     2161 cgtgcaccat ctcaatttct ttcatttata catcgttttg ccttctttta tgtaactata
     2221 ctcctctaag tttcaatctt ggccatgtaa cctctgatct atagaatttt ttaaatgact
     2281 agaattaatg cccatctttt ttttggacct aaattcttca tgaaaatata ttacgagggc
     2341 ttattcagaa gctttggact tcttcgccag aggtttggtc aagtctccaa tcaaggttgt
     2401 cggcttgtct accttgccag aaatttacga aaagatggaa aagggtcaaa tcgttggtag
     2461 atacgttgtt gacacttcta aataagcgaa tttcttatga tttatgattt ttattattaa
     2521 ataagttata aaaaaaataa gtgtatacaa attttaaagt gactcttagg ttttaaaacg
     2581 aaaattctta ttcttgagta actctttcct gtaggtcagg ttgctttctc aggtatagca
     2641 tgaggtcgct cttattgacc acacctctac cggccggtcg aaattcccct accctatgaa
     2701 catattccat tttgtaattt cgtgtcgttt ctattatgaa tttcatttat aaagtttatg
     2761 tacaaatatc ataaaaaaag agaatctttt taagcaagga ttttcttaac ttcttcggcg
     2821 acagcatcac cgacttcggt ggtactgttg gaaccaccta aatcaccagt tctgatacct
     2881 gcatccaaaa cctttttaac tgcatcttca atggccttac cttcttcagg caagttcaat
     2941 gacaatttca acatcattgc agcagacaag atagtggcga tagggttgac cttattcttt
     3001 ggcaaatctg gagcagaacc gtggcatggt tcgtacaaac caaatgcggt gttcttgtct
     3061 ggcaaagagg ccaaggacgc agatggcaac aaacccaagg aacctgggat aacggaggct
     3121 tcatcggaga tgatatcacc aaacatgttg ctggtgatta taataccatt taggtgggtt
     3181 gggttcttaa ctaggatcat ggcggcagaa tcaatcaatt gatgttgaac cttcaatgta
     3241 ggaaattcgt tcttgatggt ttcctccaca gtttttctcc ataatcttga agaggccaaa
     3301 acattagctt tatccaagga ccaaataggc aatggtggct catgttgtag ggccatgaaa
     3361 gcggccattc ttgtgattct ttgcacttct ggaacggtgt attgttcact atcccaagcg
     3421 acaccatcac catcgtcttc ctttctctta ccaaagtaaa tacctcccac taattctctg
     3481 acaacaacga agtcagtacc tttagcaaat tgtggcttga ttggagataa gtctaaaaga
     3541 gagtcggatg caaagttaca tggtcttaag ttggcgtaca attgaagttc tttacggatt
     3601 tttagtaaac cttgttcagg tctaacacta cctgtacccc atttaggacc acccacagca
     3661 cctaacaaaa cggcatcagc cttcttggag gcttccagcg cctcatctgg aagtgggaca
     3721 cctgtagctt cgatagcagc accaccaatt aaatgatttt cgaaatcgaa cttgacattg
     3781 gaacgaacat cagaaatagc tttaagaacc ttaatggctt cggctgtgat ttcttgacca
     3841 acgtggtcac ctggcaaaac gacgatcttc ttaggggcag acattagaat ggtatatcct
     3901 tgaaatatat atatatattg ctgaaatgta aaaggtaaga aaagttagaa agtaagacga
     3961 ttgctaacca cctattggaa aaaacaatag gtccttaaat aatattgtca acttcaagta
     4021 ttgtgatgca agcatttagt catgaacgct tctctattct atatgaaaag ccggttccgg
     4081 cgctctcacc tttccttttt ctcccaattt ttcagttgaa aaaggtatat gcgtcaggcg
     4141 acctctgaaa ttaacaaaaa atttccagtc atcgaatttg attctgtgcg atagcgcccc
     4201 tgtgtgttct cgttatgttg aggaaaaaaa taatggttgc taagagattc gaactcttgc
     4261 atcttacgat acctgagtat tcccacagtt gggggatctc gactctagct agaggatcaa
     4321 ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     4381 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     4441 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4501 ataacttcgt ataatgtatg ctatacgaag ttattaggtc tgaagaggag tttacgtcca
     4561 gccaagctag cttggctgca ggtcgagcgg ccgcgatccg gaacccttaa tataacttcg
     4621 tataatgtat gctatacgaa gttatcagct gcattaatga atcggccaac gcgcggggag
     4681 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4741 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4801 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4861 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4921 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4981 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5041 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5101 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5161 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5221 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5281 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5341 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5401 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5461 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5521 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5581 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5641 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5701 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5761 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5821 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5881 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5941 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6001 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6061 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6121 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6181 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6241 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6301 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6361 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6421 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6481 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6541 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6601 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
     6661 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
     6721 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     6781 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     6841 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata acgcatttaa
     6901 gcataaacac gcactatgcc gttcttctca tgtatatata tatacaggca acacgcagat
     6961 ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc gcgttgcatt ttcggaagcg
     7021 ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt cctattctct agctagaaag
     7081 tataggaact tcagagcgct tttgaaaacc aaaagcgctc tgaagacgca ctttcaaaaa
     7141 accaaaaacg caccggactg taacgagcta ctaaaatatt gcgaataccg cttccacaaa
     7201 cattgctcaa aagtatctct ttgctatata tctctgtgct atatccctat ataacctacc
     7261 catccacctt tcgctccttg aacttgcatc taaactcgac ctctacattt tttatgttta
     7321 tctctagtat tactctttag acaaaaaaat tgtagtaaga actattcata gagtgaatcg
     7381 aaaacaatac gaaaatgtaa acatttccta tacgtagtat atagagacaa aatagaagaa
     7441 accgttcata attttctgac caatgaagaa tcatcaacgc tatcactttc tgttcacaaa
     7501 gtatgcgcaa tccacatcgg tatagaatat aatcggggat gcctttatct tgaaaaaatg
     7561 cacccgcagc ttcgctagta atcagtaaac gcgggaagtg gagtcaggct ttttttatgg
     7621 aagagaaaat agacaccaaa gtagccttct tctaacctta acggacctac agtgcaaaaa
     7681 gttatcaaga gactgcatta tagagcgcac aaaggagaaa aaaagtaatc taagatgctt
     7741 tgttagaaaa atagcgctct cgggatgcat ttttgtagaa caaaaaagaa gtatagattc
     7801 tttgttggta aaatagcgct ctcgcgttgc atttctgttc tgtaaaaatg cagctcagat
     7861 tctttgtttg aaaaattagc gctctcgcgt tgcatttttg ttttacaaaa atgaagcaca
     7921 gattcttcgt tggtaaaata gcgctttcgc gttgcatttc tgttctgtaa aaatgcagct
     7981 cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgttcta caaaatgaag
     8041 cacagatgct tcgttgct
//
