LOCUS       p20StAD                 8218 bp    DNA     circular UNA 29-DEC-2019
DEFINITION  Gibson Assembly of Frag 1 TDH3 promoter PCR Product - Frag 2 GAL4
            AD MCS - Frag 3 TDH1 terminator - Frag 4 Leu into Digestion of E-V
            1 p20_int - Fragment 2.
ACCESSION   urn.local...5g-f2ut9nx
VERSION     urn.local...5g-f2ut9nx
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     misc_feature    628..633
                     /Recognition_pattern="T^CTAGA"
                     /annotation_group="TCTAGA"
                     /label="XbaI"
                     /note="Geneious type: restriction site"
     origin          647^648
                     /label="origin"
     misc_feature    complement(658..677)
                     /Sequence="TAATACGACTCACTATAGGG"
                     /Hairpin_Tm="None"
                     /Tm="50.3"
                     /Self_Dimer_Tm="None"
                     /%GC="40.0"
                     /created_by="primer3"
                     /label="Universal T7"
                     /note="Geneious type: primer_bind_reverse"
     primer_bind     complement(684..701)
                     /Sequence="TGTAAAACGACGGCCAGT"
                     /Hairpin_Tm="None"
                     /Tm="56.8"
                     /Self_Dimer_Tm="None"
                     /%GC="50.0"
                     /modified_by="User"
                     /label="universal M13-20"
     misc_feature    768..773
                     /Recognition_pattern="CAG^CTG"
                     /annotation_group="CAGCTG"
                     /label="PvuII"
                     /note="Geneious type: restriction site"
     promoter        1571..1676
                     /created_by="User"
                     /label="ampR promoter"
     CDS             1677..2537
                     /created_by="User"
                     /modified_by="User"
                     /label="Amp resistance"
     rep_origin      2707..3296
                     /created_by="User"
                     /modified_by="User"
                     /label="prokaryote ori"
     misc_feature    3530..3535
                     /Recognition_pattern="CAG^CTG"
                     /annotation_group="CAGCTG"
                     /label="PvuII"
                     /note="Geneious type: restriction site"
     primer_bind     3665..3684
                     /Sequence="AGCGGATAACAATTTCACAC"
                     /Hairpin_Tm="None"
                     /Tm="54.3"
                     /Self_Dimer_Tm="None"
                     /%GC="40.0"
                     /modified_by="User"
                     /label="universal PUC REV"
     primer_bind     3689..3708
                     /Sequence="AACAGCTATGACCATGATTA"
                     /Hairpin_Tm="None"
                     /Tm="51.6"
                     /Self_Dimer_Tm="None"
                     /%GC="35.0"
                     /created_by="primer3"
                     /label="Universal M13 R"
     unsure          3749..4417
                     /created_by="User"
                     /modified_by="User"
                     /label="locus 20 homology UP"
     misc_feature    4417
                     /label="OL"
     source          <4418..>5084
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /label="source synthetic DNA construct"
     promoter        4418..5084
                     /gene="S. cerevisiae TDH3"
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase"
                     /label="GAP promoter"
     domain          5091..>5498
                     /note="encodes 5' region of the Gal 4 activation domain
                     polypeptide; stop codon is located downstream within the
                     multiple cloning site"
                     /codon_start=1
                     /transl_table=11
                     /product="Gal 4 activation domain polypeptide"
                     /protein_id="AAB39921.1"
                     /db_xref="GI:915411"
                     /translation="MDKAELIPEPPKKKRKVELGTAANFNQSGNIADSSLSFTFTNSS
                     NGPNLITTQTNSQALSQPIASSNVHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYN
                     AFGITTGMFNTTTMDDVYNYLFDDEDTPPNPKKE"
                     /modified_by="User"
                     /label="Gal 4 activation domain"
     terminator      5696..5892
                     /created_by="User"
                     /modified_by="User"
                     /label="TDH1 term"
     misc_feature    complement(5900..>5910)
                     /label="pMSs BstZ17i"
     misc_feature    complement(<5911..5955)
                     /note="Lox Site"
                     /label="Lox Site"
     misc_feature    complement(5995..6054)
                     /note="Lox Site"
                     /label="Lox Site"
     unsure          6279..6287
                     /created_by="User"
                     /label
     ORF             complement(6754..7848)
                     /codon_start=1
                     /transl_table=11
                     /product="LEU2"
                     /protein_id="AAB39920.1"
                     /db_xref="GI:915410"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENH
                     LIGGAAIDATGVPLPDEALEASKKVDAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQL
                     YANLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSE
                     QYTVPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLK
                     VQHQLIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPD
                     KNTAFGLYEPCHGSAPDLPKNKVDPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLD
                     AGIRTGDLGGSNSTTEVGDAVAEEVKKILA"
                     /modified_by="User"
                     /label="LEU2 ORF"
     unsure          8195..647
                     /created_by="User"
                     /modified_by="User"
                     /label="locus 20 homology DOWN"
ORIGIN      
        1 catatacaag cactaagaag aacaatacag aactctacac ggtattattg tgctacaagc
       61 tcgagtaaaa ccgagtgttt tgacgatact aacgttgtta agaaagtaac ttgttatcaa
      121 actcattacc aacttgtgat taattggtga ataatatgat aattgtcgaa attccattgt
      181 tggtaaagcc tataatatta tgtatacaga ttatactaga aattctctcg agaatataag
      241 aatccccaaa attgaatcgg tatttctaca tactaatatt accattactt ctcctttcgt
      301 tttatatgtt tcattcctat tacattatcg atctttgcat ttcagcttcc attatatttg
      361 atgtctgttt tatgtcccca cgttacaccg catgtgacag tatactagta acatgagtgc
      421 taccgaatag atgacatttt agactttcat tccaacaact tggttgacag aatgttacgt
      481 accctatatc taatctatat gaggcctgaa tctaactgaa aggtggaatt tcagtaattt
      541 atcaagcttt aataagtttg ggtagtttaa ctgtgcaaaa aggtatttac cttacatact
      601 gaatcttgtc tgtttggtag cggctgctct agaactagtg gatcgatccc caattcgccc
      661 tatagtgagt cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
      721 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      781 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
      841 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
      901 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
      961 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     1021 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggt gggccatcgc
     1081 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     1141 tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga
     1201 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     1261 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     1321 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     1381 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     1441 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     1501 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     1561 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     1621 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     1681 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     1741 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     1801 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     1861 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     1921 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     1981 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     2041 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     2101 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     2161 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     2221 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     2281 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     2341 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     2401 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     2461 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     2521 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     2581 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     2641 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     2701 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     2761 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     2821 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     2881 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     2941 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     3001 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     3061 gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc gccacgcttc
     3121 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     3181 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     3241 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     3301 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     3361 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     3421 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     3481 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     3541 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     3601 ctcattaggc accccaggct ttacacttta tgcttccgcg gctcgtatgt tgtgtggaat
     3661 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccactagtcc
     3721 gaggccgcca ggcgccttta tatcatataa ttaagacaca aaaggataaa acaaaggtgt
     3781 taactattct gcatactcac tatcgtaaac tgtcctgcaa atcgtgtaaa tatgtatttc
     3841 attttttttg cagtgaaaaa aggcatgtaa aataccgcat caagtaactc tactccgcct
     3901 gtggtttcaa gactaacggc ttgagacaaa atgggaagaa atgattgcag aaaagccata
     3961 tgtgtaatag caaaaagctg gatagtgctt accagatgtt taccttaatt tcttggtgaa
     4021 ttagagaagt acagaagttt tactattaat cccaccatag aaatttgtat aggaaagtag
     4081 tttattggag ttattggata tactgtgtaa actatttctt gaaattgtaa tcttaagatg
     4141 ctcttcttat tctattaaaa atagaaaatg attttcatat ttatttattt atttatattt
     4201 tggcattact cttcatcatt tttttccctc taagaagctt cctttctttt tataaggata
     4261 acaaaaccaa aaggaatatt gggtcagatg aatggacgcg aatgcaagac agaagtccaa
     4321 atcacgtcaa gacaaagaaa gaaagaaaga aaaactaaca cattaatgta gttttaaaat
     4381 ttcaaatccg aacaacagag catagggttt cgcaaactca ttatcaatac tgccatttca
     4441 aagaatacgt aaataattaa tagtagtgat tttcctaact ttatttagtc aaaaaattag
     4501 ccttttaatt ctgctgtaac ccgtacatgc ccaaaatagg gggcgggtta cacagaatat
     4561 ataacatcgt aggtgtctgg gtgaacagtt tattcctggc atccactaaa tataatggag
     4621 cccgcttttt aagctggcat ccagaaaaaa aaagaatccc agcaccaaaa tattgttttc
     4681 ttcaccaacc atcagttcat aggtccattc tcttagcgca actacagaga acaggggcac
     4741 aaacaggcaa aaaacgggca caacctcaat ggagtgatgc aacctgcctg gagtaaatga
     4801 tgacacaagg caattgaccc acgcatgtat ctatctcatt ttcttacacc ttctattacc
     4861 ttctgctctc tctgatttgg aaaaagctga aaaaaaaggt tgaaaccagt tccctgaaat
     4921 tattccccta cttgactaat aagtatataa agacggtagg tattgattgt aattctgtaa
     4981 atctatttct taaacttctt aaattctact tttatagtta gtcttttttt tagttttaaa
     5041 acaccaagaa cttagtttcg aataaacaca cataaacaaa caaagcaaag atggataaag
     5101 cggaattaat tcccgagcct ccaaaaaaga agagaaaggt cgaattgggt accgccgcca
     5161 attttaatca aagtgggaat attgctgata gctcattgtc cttcactttc actaacagta
     5221 gcaacggtcc gaacctcata acaactcaaa caaattctca agcgctttca caaccaattg
     5281 cctcctctaa cgttcatgat aacttcatga ataatgaaat cacggctagt aaaattgatg
     5341 atggtaataa ttcaaaacca ctgtcacctg gttggacgga ccaaactgcg tataacgcgt
     5401 ttggaatcac tacagggatg tttaatacca ctacaatgga tgatgtatat aactatctat
     5461 tcgatgatga agatacccca ccaaacccaa aaaaagagat ctgtatggct tacccatacg
     5521 atgttccaga ttacgctagc ttgggtggtc atatggccat ggaggccccg gggatccgaa
     5581 ttcgagctcg agagatctat gaatcgtaga tactgaaaaa ccccgcaagt tcacttcaac
     5641 tgtgcatcgt gcaccatctc aatttctttc atttatacat cgttttgcct tctttataaa
     5701 gcaatcttga tgaggataat gatttttttt tgaatataca taaatactac cgtttttctg
     5761 ctagattttg tgaagacgta aataagtaca tattactttt taagccaaga caagattaag
     5821 cattaacttt acccttttct cttctaagtt tcaatactag ttatcactgt ttaaaagtta
     5881 tggcgagaac gtcggcggtt acaaatttta gagggaccta ataacttcgt atagcataca
     5941 ttatacgaag ttatattaag ggttccggat cgcggccgct cgacctgcag ccaagctagc
     6001 ggacgtaaac tcctcttcag acctaataac ttcgtatagc atacattata cgaagttata
     6061 ttaagggtta ttgaatatga tcggaattgg tcgagggacc taataacttc gtatagcata
     6121 cattatacga agttatatta agggttccgg atcgcggccg ctcgacctgc agccaggcta
     6181 gcttggctgg acgtaaactc ctcttcagac ctaataactt cgatagcata cattatacga
     6241 agttatatta agggttattg atatgatcgg aattggtgca ctacgtgctt aaggccgttt
     6301 ctgacagagt aaaattcttg agggaacttt caccattatg ggaaatggtt caagaaggta
     6361 ttgacttaaa ctccatcaaa tggtcaggtc attgagtgtt ttttatttgt tgtatttttt
     6421 tttttttaga gaaaatcctc caatatcaaa ttaggaatcg tagtttcatg attttctgtt
     6481 acacctaact ttttgtgtgg tgccctcctc cttgtcaata ttaatgttaa agtgcaattc
     6541 tttttcctta tcacgttgag ccattagtat caatttgctt acctgtattc ctttactatc
     6601 ctcctttttc tccttcttga taaatgtatg tagattgcgt atatagtttc gtctacccta
     6661 tgaacatatt ccattttgta atttcgtgtc gtttctatta tgaatttcat ttataaagtt
     6721 tatgtacaaa tatcataaaa aaagagaatc tttttaagca aggattttct taacttcttc
     6781 ggcgacagca tcaccgactt cggtggtact gttggaacca cctaaatcac cagttctgat
     6841 acctgcatcc aaaacctttt taactgcatc ttcaatggcc ttaccttctt caggcaagtt
     6901 caatgacaat ttcaacatca ttgcagcaga caagatagtg gcgatagggt caaccttatt
     6961 ctttggcaaa tctggagcag aaccgtggca tggttcgtac aaaccaaatg cggtgttctt
     7021 gtctggcaaa gaggccaagg acgcagatgg caacaaaccc aaggaacctg ggataacgga
     7081 ggcttcatcg gagatgatat caccaaacat gttgctggtg attataatac catttaggtg
     7141 ggttgggttc ttaactagga tcatggcggc agaatcaatc aattgatgtt gaaccttcaa
     7201 tgtaggaaat tcgttcttga tggtttcctc cacagttttt ctccataatc ttgaagaggc
     7261 caaaacatta gctttatcca aggaccaaat aggcaatggt ggctcatgtt gtagggccat
     7321 gaaagcggcc attcttgtga ttctttgcac ttctggaacg gtgtattgtt cactatccca
     7381 agcgacacca tcaccatcgt cttcctttct cttaccaaag taaatacctc ccactaattc
     7441 tctgacaaca acgaagtcag tacctttagc aaattgtggc ttgattggag ataagtctaa
     7501 aagagagtcg gatgcaaagt tacatggtct taagttggcg tacaattgaa gttctttacg
     7561 gatttttagt aaaccttgtt caggtctaac actaccggta ccccatttag gaccacccac
     7621 agcacctaac aaaacggcat caaccttctt ggaggcttcc agcgcctcat ctggaagtgg
     7681 gacacctgta gcatcgatag cagcaccacc aattaaatga ttttcgaaat cgaacttgac
     7741 attggaacga acatcagaaa tagctttaag aaccttaatg gcttcggctg tgatttcttg
     7801 accaacgtgg tcacctggca aaacgacgat cttcttaggg gcagacatag gggcagacat
     7861 tagaatggta tatccttgaa atatatatat atattgctga aatgtaaaag gtaagaaaag
     7921 ttagaaagta agacgattgc taaccaccta ttggaaaaaa caataggtcc ttaaataata
     7981 ttgtcaactt caagtattgt gatgcaagca tttagtcatg aacgcttctc tattctatat
     8041 gaaaagccgg ttccggcctc tcacctttcc tttttctccc aatttttcag ttgaaaaagg
     8101 tatatgcgtc aggcgacctc tgaaattaac aaaaaatttc cagtcatcga atttgattct
     8161 gtgcgatagc gcccctgtgt gttctcgtta tgttgtacca atggaaggtc gggatgag
//
