LOCUS       sequence-209173-        7751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  destabilizing domains for yeast (S. cerevisiae) derived from the 
            human estrogen receptor ligand binding domain. This ERdd can be 
            stabilized with 10 uM 4-hydroxytamoxifen (4-OHT, OHTAM)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7751)
  TITLE     pScDD2_ERdd
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7751)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pScDD2_ERdd
FEATURES             Location/Qualifiers
     source          1..7751
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      11..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(98..117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     308..329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     593..615
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     607..624
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     608..624
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     634..653
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        634..652
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1811..2524
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1811..2524
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      3427..3614
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(3482..3506)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     gene            3667..5023
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     promoter        3667..4010
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     CDS             4011..4820
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4078..4097)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      4826..5023
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(5072..5083)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             5549..5560
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(5570..5590)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5570..5588)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5609..5625)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5609..5625)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5622..5644)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5633..5649
                     /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(join(5657..5663,5664..5681,5682..5687))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5702..5723
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5840..5857)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6011..6599)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6091..6110)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6770..7561,7562..7630))
                     /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     7393..7412
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7631..7735)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
       61 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
      121 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
      181 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
      241 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
      301 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
      361 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcca ttcgccattc
      481 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      541 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      601 cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg actcactata gggcgaattg
      661 ggtaccattt tatacactct ggtcctttaa ctggcaaacc ttcgggcgta atgcccaatt
      721 tttcgccttt gtcttttgcc tttttcactt cacgtgcttc tggtacatac ttgcaattta
      781 tacagtgatg accgctgaat ttgtatcttc catagcatct agcacatact cgatttttac
      841 cactccaatc tttataaaaa tacttgattc cctttctggg acaagcaaca cagtgtttta
      901 gattcttttt ttgtgatatt ttaagctgtt ctcccacaca gcagcctcga catgatttca
      961 cttctatttt gttgccaggc aagaaatttt tatggccttc tatcgtaagc ccatatacag
     1021 tactctcacc ctggaaatca tccgtgaagc tgaaatatac gggttccctt tttataattg
     1081 gcggaacttc tcttgttttg tgaccacttc gacaatatga caaaacattc tgtgaagttg
     1141 ttcccccagc aacattacag tcgtatctcg agttggaaat ctaaaagtgg gttacccaat
     1201 actaccaatg gtgcggtcat aattgcttgc tctttctttt gctgttatct ttggttctac
     1261 cctgcacaag ataaactgag atgactacct aattagacat ggcatgccta taagtaaaga
     1321 gaattgggct cgaagaataa ttttcaagcc tgccctcatc acgtacgacg acactgcgac
     1381 tcatccatgt gaaaattatc ggcatctgca aaaaaagttt taacttccac aggtaatatt
     1441 ggcatgatgc gaaattggac gtaagtatct ctgaagtgca gccgattggg cgtgcgactc
     1501 acccactcag gacatgatct cagtagcggg ttcgataagg cgatgacagc gcaaatgccg
     1561 cttactggaa gtacagaacc cgctccctta ggggcaccca ccccagcacg ctggagcgtt
     1621 aaaccggtgt gtcggaatta gtaagcggac atcccttccg ctgggctcgc catcgcagat
     1681 atatatataa gaagatggtt ttgggcaaat gtttagctgt aactatgttg cggaaaaaca
     1741 ggcaagaaag caatcgcgca aacaaataaa acataattaa tttatactgc agcccggggg
     1801 atccaaaaaa atgtctaaag gtgaagaatt attcactggt gttgtcccaa ttttggttga
     1861 attagatggt gatgttaatg gtcacaaatt ttctgtctcc ggtgaaggtg aaggtgatgc
     1921 tacttacggt aaattgacct taaaatttat ttgtactact ggtaaattgc cagttccatg
     1981 gccaacctta gtcactactt taacttatgg tgttcaatgt ttttctagat acccagatca
     2041 tatgaaacaa catgactttt tcaagtctgc catgccagaa ggttatgttc aagaaagaac
     2101 tatttttttc aaagatgacg gtaactacaa gaccagagct gaagtcaagt ttgaaggtga
     2161 taccttagtt aatagaatcg aattaaaagg tattgatttt aaagaagatg gtaacatttt
     2221 aggtcacaaa ttggaataca actataactc tcacaatgtt tacatcatgg ctgacaaaca
     2281 aaagaatggt atcaaagtta acttcaaaat tagacacaac attgaagatg gttctgttca
     2341 attagctgac cattatcaac aaaatactcc aattggtgat ggtccagtct tgttaccaga
     2401 caaccattac ttatccactc aatctgcctt atccaaagat ccaaacgaaa agagagacca
     2461 catggtcttg ttagaatttg ttactgctgc tggtattacc catggtatgg atgaattgta
     2521 caaaactagt ggtagaggag aaggtcaaat ggctctgtcc ttgactgctg accaaatggt
     2581 ttctgctttg ttggacgctg aaccaccaat cttgtactct gaatacgacc caactagacc
     2641 attctctgaa gcttctatga tgggtttgtt gactaacttg gctgacagag aattggttca
     2701 catgatcaac tgggctaaga gagttccagg tttcgttgac ttgactttgc acgaccaagt
     2761 tcacttgttg ggatgtgctt ggttggaaat cttgatgatc ggtttggttt ggagatctat
     2821 ggaacaccca ggtaagttgt tgttcgctcc aaacttgttg ttggacagaa accaaggtaa
     2881 gtgtgttgaa ggtatggttg aaatcttcga catgttgttg gctacttctt ctggattcag
     2941 aatgatgaac ttgcaaggtg aagaattcgt ttgtttgaag tctatcatct tgttgaactc
     3001 tggtgtttac actttcttgt cttctacttt gaagtctttg gaagaaaagg accacatcca
     3061 cagagttttg gacaagatca ctgacacttt gatccacttg atggctaagg ctggtttgac
     3121 tttgcaacaa caacaccaaa gattggctca attgttgttg atcttgtctc acatcagaca
     3181 catgtctaac aagggtatgg aacacttgta ctctatgaag tgtaagagcg ttgttccatt
     3241 gtacgacttg ttgttggaaa tgttggacgc tcacagattg taaccgcgga agctttggac
     3301 ttcttcgcca gaggtttggt caagtctcca atcaaggttg tcggcttgtc taccttgcca
     3361 gaaatttacg aaaagatgga aaagggtcaa atcgttggta gatacgttgt tgacacttct
     3421 aaataagcga atttcttatg atttatgatt tttattatta aataagttgt aaaaaaaata
     3481 agtgtataca aattttaaag tgactcttag gttttaaaac gaaaattctt attcttgagt
     3541 aactctttcc tgtaggtcag gttgctttct caggtatagc atgaggtcgc tcttattgac
     3601 cacacctcta ccggcatgcc ctgcaggatc gatatcgcct cgtccccgcc gggtcacccg
     3661 gccagcgaca tggaggccca gaataccctc cttgacagtc ttgacgtgcg cagctcaggg
     3721 gcatgatgtg actgtcgccc gtacatttag cccatacatc cccatgtata atcatttgca
     3781 tccatacatt ttgatggccg cacggcgcga agcaaaaatt acggctcctc gctgcagacc
     3841 tgcgagcagg gaaacgctcc cctcacagac gcgttgaatt gtccccacgc cgcgcccctg
     3901 tagagaaata taaaaggtta ggatttgcca ctgaggttct tctttcatat acttcctttt
     3961 aaaatcttgc taggatacag ttctcacatc acatccgaac ataaacaacc atgggtaagg
     4021 aaaagactca cgtttcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     4081 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga
     4141 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     4201 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     4261 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggcaaaacag
     4321 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     4381 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     4441 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     4501 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataagcttt
     4561 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     4621 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     4681 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     4741 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     4801 tgctcgatga gtttttctaa tcagtactga caataaaaag attcttgttt tcaagaactt
     4861 gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg ttagcgtgat
     4921 ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg cgcagaaagt
     4981 aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga ttcgatacta
     5041 acggcggccg cgtaaattga cattggactt ttcttccttc aatgatttcc tccctagctg
     5101 acctggtcgt cacagtagct gacataccaa gagatcttga aatatggaca attccgtcca
     5161 taatggatga ataaacagtt tgaattgcta ttttataaga ttcagggcct tcgccctttt
     5221 tcacaacgta cccatctgag tcgatcaagc cggctaagaa tgcttcacga acttctatat
     5281 gctcgccgta cataaattca gggatttgct tctctccatc aagatccctt ttaaacttta
     5341 aaattgtgac agctttccag aatggattat ttttcctcaa attccttgtc ttcctgtttt
     5401 catctggacc atctccataa tgaagcctta catgtttggc acgtagcgga acgtgatcgt
     5461 cacaaaccgt aaggtagaga ccccagattt tcgcattttc tcttaaactc tccattagct
     5521 taggatccaa gctatctact gagatttctg ggagctccag cttttgttcc ctttagtgag
     5581 ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     5641 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     5701 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     5761 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     5821 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     5881 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     5941 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6001 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6061 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6121 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6181 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6241 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6301 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6361 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6421 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     6481 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     6541 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     6601 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     6661 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     6721 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6781 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6841 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6901 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6961 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7021 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7081 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7141 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7201 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7261 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7321 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7381 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7441 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7501 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7561 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7621 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7681 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     7741 ttccccgaaa a
//
