LOCUS       Exported                8905 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast expression vector for human estrogen receptor alpha.
ACCESSION   .
VERSION     .
KEYWORDS    pG/ER(G)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8905)
  AUTHORS   Liu JW, Picard D
  TITLE     Bioactive steroids as contaminants of the common carbon source 
            galactose.
  JOURNAL   FEMS Microbiol Lett. 1998 Feb 15;159(2):167-71.
  PUBMED    9503609
REFERENCE   2  (bases 1 to 8905)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8905
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        770..1413
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase"
     primer_bind     1306..1329
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             2320..3213
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     terminator      3359..3629
                     /gene="S. cerevisae PGK1"
                     /label=PGK1 terminator
                     /note="transcription terminator for 3-phosphoglycerate 
                     kinase gene"
     primer_bind     complement(3709..3728)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3927..4028
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     CDS             4029..4703
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     primer_bind     complement(4799..4816)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4799..4815)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4808..4830)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5024..5043)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5143..5165
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5203..5221)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5289..5393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5394..6254
                     /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(5612..5631)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6425..7013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6914..6933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7167..7184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7301..7322
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7337..7367
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7375..7391
                     /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     7380..7402
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7399..7415
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7399..7415
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    8099..8146
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
        1 agctcagatt ctttgtttga aaaattagcg ctctcgcgtt gcatttttgt tctacaaaat
       61 gaagcacaga tgcttcgtta acaaagatat gctattgaag tgcaagatgg aaacgcagaa
      121 aatgaaccgg ggatgcgacg tgcaagatta cctatgcaat agatgcaata gtttctccag
      181 gaaccgaaat acatacattg tcttccgtaa agcgctagac tatatattat tatacaggtt
      241 caaatatact atctgtttca gggaaaactc ccaggttcgg atgttcaaaa ttcaatgatg
      301 ggtaacaagt acgatcgtaa atctgtaaaa cagtttgtcg gatattaggc tgtatctcct
      361 caaagcgtat tcgaatatca ttgagaagct gcagctattc atgcaactgc taaatggtct
      421 actgctcaat taaagaaaac acaggcagag caggctgccc gggcaaaagc tgcagcgtca
      481 catcggataa taatgatggc agccattgta gaagtgcctt ttgcatttct agtctctttc
      541 tcggtctagc tagttttact acatcgcgaa gatagaatct tagatcacac tgcctttgct
      601 gagctggatc aatagagtaa caaaagagtg gtaaggcctc gttaaaggac aaggacctga
      661 gcggaagtgt atcgtacagt agacggagta tactagtata gtctatagtc cgtggaattc
      721 tcatgtttga cagcttatca tcgataagct tggctgcagg tcgagtttat cattatcaat
      781 actgccattt caaagaatac gtaaataatt aatagtagtg attttcctaa ctttatttag
      841 tcaaaaaatt agccttttaa ttctgctgta acccgtacat gcccaaaata gggggcgggt
      901 tacacagaat atataacatc gtaggtgtct gggtgaacag tttattcctg gcatccacta
      961 aatataatgg agcccgcttt ttaagctggc atccagaaaa aaaaagaatc ccagcaccaa
     1021 aatattgttt tcttcaccaa ccatcagttc ataggtccat tctcttagcg caactacaga
     1081 gaacaggggc acaaacaggc aaaaaacggg cacaacctca atggagtgat gcaacctgcc
     1141 tggagtaaat gatgacacaa ggcaattgac ccacgcatgt atctatctca ttttcttaca
     1201 ccttctatta ccttctgctc tctctgattt ggaaaaagct gaaaaaaaag gttgaaacca
     1261 gttccctgaa attattcccc tacttgacta ataagtatat aaagacggta ggtattgatt
     1321 gtaattctgt aaatctattt cttaaacttc ttaaattcta cttttatagt tagtcttttt
     1381 tttagtttta aaacaccaag aacttagttt cgacggatcc ggaccatgac catgaccctc
     1441 cacaccaaag catctgggat ggccctactg catcagatcc aagggaacga gctggagccc
     1501 ctgaaccgtc cgcagctcaa gatccccctg gagcggcccc tgggcgaggt gtacctggac
     1561 agcagcaagc ccgccgtgta caactacccc gagggcgccg cctacgagtt caacgccgcg
     1621 gccgccgcca acgcgcaggt ctacggtcag accggcctcc cctacggccc cgggtctgag
     1681 gctgcggcgt tcggctccaa cggcctgggg ggtttccccc cactcaacag cgtgtctccg
     1741 agcccgctga tgctactgca cccgccgccg cagctgtcgc ctttcctgca gccccacggc
     1801 cagcaggtgc cctactacct ggagaacgag cccagcggct acacggtgcg cgaggccggc
     1861 ccgccggcat tctacaggcc aaattcagat aatcgacgcc agggtggcag agaaagattg
     1921 gccagtacca atgacaaggg aagtatggct atggaatctg ccaaggagac tcgctactgt
     1981 gcagtgtgca atgactatgc ttcaggctac cattatggag tctggtcctg tgagggctgc
     2041 aaggccttct tcaagagaag tattcaagga cataacgact atatgtgtcc agccaccaac
     2101 cagtgcacca ttgataaaaa caggaggaag agctgccagg cctgccggct ccgcaaatgc
     2161 tacgaagtgg gaatgatgaa aggtgggata cgaaaagacc gaagaggagg gagaatgttg
     2221 aaacacaagc gccagagaga tgatggggag ggcaggggtg aagtggggtc tgctggagac
     2281 atgagagctg ccaacctttg gccaagcccg ctcatgatca aacgctctaa gaagaacagc
     2341 ctggccttgt ccctgacggc cgaccagatg gtcagtgcct tgttggatgc tgagcccccc
     2401 atactctatt ccgagtatga tcctaccaga cccttcagtg aagcttcgat gatgggctta
     2461 ctgaccaacc tggcagacag ggagctggtt cacatgatca actgggcgaa gagggtgcca
     2521 ggctttgtgg atttgaccct ccatgatcag gtccaccttc tagaatgtgc ctggctagag
     2581 atcctgatga ttggtctcgt ctggcgctcc atggagcacc cagggaagct actgtttgct
     2641 cctaacttgc tcttggacag gaaccaggga aaatgtgtag agggcatggt ggagatcttc
     2701 gacatgctgc tggctacatc atctcggttc cgcatgatga atctgcaggg agaggagttt
     2761 gtgtgcctca aatctattat tttgcttaat tctggagtgt acacatttct gtccagcacc
     2821 ctgaagtctc tggaagagaa ggaccatatc caccgagtcc tggacaagat cacagacact
     2881 ttgatccacc tgatggccaa ggcaggcctg accctgcagc agcagcacca gcggctggcc
     2941 cagctcctcc tcatcctctc ccacatcagg cacatgagta acaaaggcat ggagcatctg
     3001 tacagcatga agtgcaagaa cgtggtgccc ctctatgacc tgctgctgga gatgctggac
     3061 gcccaccgcc tacatgcgcc cactagccgt ggaggggcat ccgtggagga gacggaccaa
     3121 agccacttgg ccactgcggg ctctacttca tcgcattcct tgcaaaagta ttacatcacg
     3181 ggggaggcag agggtttccc tgccacagtc tgagagctcc ctggcggaat tcggatccgt
     3241 cgacctagat aagtaatgat ccgggatctc ccatgtctct actggtggtg gtgcttcttt
     3301 ggaattattg gaaggtaagg aattgccagg tgttgctttc ttatccgaaa agaaataaat
     3361 tgaattgaat tgaaatcgat agatcaattt ttttcttttc tctttcccca tcctttacgc
     3421 taaaataata gtttatttta ttttttgaat attttttatt tatatacgta tatatagact
     3481 attatttatc ttttaatgat tattaagatt tttattaaaa aaaaattcgc tcctctttta
     3541 atgcctttat gcagtttttt tttcccattc gatatttcta tgttcgggtt cagcgtattt
     3601 taagtttaat aactcgaaaa ttctgcgttc gttaaagctc cggatcgatc ctctacgccg
     3661 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
     3721 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
     3781 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac
     3841 cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc
     3901 aggagtcgca taagggagag cgtcgaaatt cggtcgaaaa aagaaaagga gagggccaag
     3961 agggagggca ttggtgacta ttgagcacgt gagtatacgt gattaagcac acaaaggcag
     4021 cttggagtat gtctgttatt aatttcacag gtagttctgg tccattggtg aaagtttgcg
     4081 gcttgcagag cacagaggcc gcagaatgtg ctctagattc cgatgctgac ttgctgggta
     4141 ttatatgtgt gcccaataga aagagaacaa ttgacccggt tattgcaagg aaaatttcaa
     4201 gtcttgtaaa agcatataaa aatagttcag gcactccgaa atacttggtt ggcgtgtttc
     4261 gtaatcaacc taaggaggat gttttggctc tggtcaatga ttacggcatt gatatcgtcc
     4321 aactgcatgg agatgagtcg tggcaagaat accaagagtt cctcggtttg ccagttatta
     4381 aaagactcgt atttccaaaa gactgcaaca tactactcag tgcagcttca cagaaacctc
     4441 attcgtttat tcccttgttt gattcagaag caggtgggac aggtgaactt ttggattgga
     4501 actcgatttc tgactgggtt ggaaggcaag agagccccga aagcttacat tttatgttag
     4561 ctggtggact gacgccagaa aatgttggtg atgcgcttag attaaatggc gttattggtg
     4621 ttgatgtaag cggaggtgtg gagacaaatg gtgtaaaaga ctctaacaaa atagcaaatt
     4681 tcgtcaaaaa tgctaagaaa taggttatta ctgagtagta tttatttaag tattgtttgt
     4741 gcacttgcct gcaggccttt tgaaaagcaa gcataaaaga tctgcatgca agcttggcac
     4801 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     4861 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     4921 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     4981 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     5041 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5101 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5161 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat
     5221 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5281 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5341 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5401 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5461 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5521 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5581 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5641 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5701 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5761 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5821 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5881 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5941 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6001 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6061 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6121 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6181 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6241 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6301 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6361 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6421 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6481 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6541 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6601 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6661 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6721 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6781 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6841 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6901 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6961 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7021 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7081 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     7141 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     7201 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7261 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     7321 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     7381 gcggataaca atttcacaca ggaaacagct atgacatgat tacgaattct gaaccagtcc
     7441 taaaacgagt aaataggacc ggcaattctt caagcaataa acaggaatac caattattaa
     7501 aagataactt agtcagatcg tacaataaag ctttgaagaa aaatgcgcct tattcaatct
     7561 ttgctataaa aaatggccca aaatctcaca ttggaagaca tttgatgacc tcatttcttt
     7621 caatgaaggg cctaacggag ttgactaatg ttgtgggaaa ttggagcgat aagcgtgctt
     7681 ctgccgtggc caggacaacg tatactcatc agataacagc aatacctgat cactacttcg
     7741 cactagtttc tcggtactat gcatatgatc caatatcaaa ggaaatgata gcattgaagg
     7801 atgagactaa tccaattgag gagtggcagc atatagaaca gctaaagggt agtgctgaag
     7861 gaagcatacg ataccccgca tggaatggga taatatcaca ggaggtacta gactaccttt
     7921 catcctacat aaatagacgc atataagtac gcatttaagc ataaacacgc actatgccgt
     7981 tcttctcatg tatatatata tacaggcaac acgcagatat aggtgcgacg tgaacagtga
     8041 gctgtatgtg cgcagctcgc gttgcatttt cggaagcgct cgttttcgga aacgctttga
     8101 agttcctatt ccgaagttcc tattctctag aaagtatagg aacttcagag cgcttttgaa
     8161 aaccaaaagc gctctgaaga cgcactttca aaaaaccaaa aacgcaccgg actgtaacga
     8221 gctactaaaa tattgcgaat accgcttcca caaacattgc tcaaaagtat ctctttgcta
     8281 tatatctctg tgctatatcc ctatataacc tacccatcca cctttcgctc cttgaacttg
     8341 catctaaact cgacctctac attttttatg tttatctcta gtattactct ttagacaaaa
     8401 aaattgtagt aagaactatt catagagtga atcgaaaaca atacgaaaat gtaaacattt
     8461 cctatacgta gtatatagag acaaaataga agaaaccgtt cataattttc tgaccaatga
     8521 agaatcatca acgctatcac tttctgttca caaagtatgc gcaatccaca tcggtataga
     8581 atataatcgg ggatgccttt atcttgaaaa aatgcacccg cagcttcgct agtaatcagt
     8641 aaacgcggga agtggagtca ggcttttttt atggaagaga aaatagacac caaagtagcc
     8701 ttcttctaac cttaacggac ctacagtgca aaaagttatc aagagactgc attatagagc
     8761 gcacaaagga gaaaaaaagt aatctaagat gctttgttag aaaaatagcg ctctcgggat
     8821 gcatttttgt agaacaaaaa agaagtatag attctttgtt ggtaaaatag cgctctcgcg
     8881 ttgcatttct gttctgtaaa aatgc
//
