LOCUS       sequence-172828-        9173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively expresses estradiol-inducible synthetic transcription
            factor in yeast. It activates transcription of promoters containing 
            Zif268 binding sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9173)
  AUTHORS   Aranda-Diaz A, Mace K, Zuleta I, Harrigan P, El-Samad H
  TITLE     Robust Synthetic Circuits for Two-Dimensional Control of Gene 
            Expression in Yeast.
  JOURNAL   ACS Synth Biol. 2016 Dec 27. doi: 10.1021/acssynbio.6b00251.
  PUBMED    27930885
REFERENCE   2  (bases 1 to 9173)
  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: "ACS Synth 
            Biol. 2016 Dec 27. doi: 10.1021/acssynbio.6b00251."
COMMENT     Sequence Label: pHES795
FEATURES             Location/Qualifiers
     source          1..9173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        569..912
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     terminator      1731..1928
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     protein_bind    2574..2592
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2595..2613
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             2942..3877
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2990..3880
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     primer_bind     complement(5720..5739)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5721..5739)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5746..5763)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5746..5762)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5755..5777)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5903..6358
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5990..6009)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6200..6221
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6714..6733)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6833..6855
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6893..6911)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6996..7100
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7101..7169,7170..7961)
                     /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(7319..7338)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8132..8720
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8621..8640
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8874..8891
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9008..9029
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9044..9049,9050..9067,9068..9074)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9082..9098
                     /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     9087..9109
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9106..9122
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9106..9122
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        9143..9161
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggccggccat ttaaatgttt aaacagaacg aaggaaggag cacagactta gattggtata
       61 tatacgcata tgtggtgttg aagaaacatg aaattgccca gtattcttaa cccaactgca
      121 cagaacaaaa acctgcagga aacgaagata aatcatgtcg aaagctacat ataaggaacg
      181 tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc acgaaaagca
      241 aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg agttagttga
      301 agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga ctgatttttc
      361 catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt ttttactctt
      421 cgaagacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact ctgcgggtgt
      481 atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg tttagcttgc
      541 ctcgtccccg ccgggtcacc cggccagcga catggaggcc cagaataccc tccttgacag
      601 tcttgacgtg cgcagctcag gggcatgatg tgactgtcgc ccgtacattt agcccataca
      661 tccccatgta taatcatttg catccataca ttttgatggc cgcacggcgc gaagcaaaaa
      721 ttacggctcc tcgctgcaga cctgcgagca gggaaacgct cccctcacag acgcgttgaa
      781 ttgtccccac gccgcgcccc tgtagagaaa tataaaaggt taggatttgc cactgaggtt
      841 cttctttcat atacttcctt ttaaaatctt gctaggatac agttctcaca tcacatccga
      901 acataaacaa ccatgacagt caacactaag acctatagtg agagagcaga aactcatgcc
      961 tcaccagtag cacagcgatt atttcgatta atggaactga agaaaaccaa tttatgtgca
     1021 tcaattgacg ttgataccac taaggaattc cttgaattaa ttgataaatt aggtccttat
     1081 gtatgcttaa tcaagactca tattgatata atcaatgatt tttcctatga atccactatt
     1141 gaaccattat tagaactttc acgtaaacat caatttatga tttttgaaga tagaaaattt
     1201 gctgatattg gtaataccgt aaagaaacaa tatattggtg gagtttataa aattagtagt
     1261 tgggcagata ttaccaatgc tcatggtgtc actgggaatg gagtggttga aggattaaaa
     1321 cagggagcta aagaaaccac caccaaccaa gagccaagag ggttattgat gttagctgaa
     1381 ttatcatcag tgggatcatt agcatatgga gaatattctc aaaaaactgt tgaaattgct
     1441 aaatccgata aggaatttgt tattggattt attgcccaac gtgatatggg tggccaagaa
     1501 gaaggatttg attggcttat tatgacacct ggagttggat tagatgataa aggtgatgga
     1561 ttaggacaac aatatagaac tgttgatgaa gttgttagca ctggaactga tattatcatt
     1621 gttggtagag gattgtttgg taaaggaaga gatccagata ttgaaggtaa aaggtataga
     1681 aatgctggtt ggaatgctta tttgaaaaag actggccaat tataatcagt actgacaata
     1741 aaaagattct tgttttcaag aacttgtcat ttgtatagtt tttttatatt gtagttgttc
     1801 tattttaatc aaatgttagc gtgatttata ttttttttcg cctcgacatc atctgcccag
     1861 atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa tcgtatgtga atgctggtcg
     1921 ctatactgct gtcgattcga tactaacgcc gccatccagt gtcgaaaacg acctcgaatt
     1981 catcgagtgg attcggcttt gggtaccggg cccggcaacc aaacccatac atcgggattc
     2041 ctataatacc ttcgttggtc tccctaacat gtaggtggcg gaggggagat atacaataga
     2101 acagatacca gacaagacat aatgggctaa acaagactac accaattaca ctgcctcatt
     2161 gatggtggta cataacgaac taatactgta gccctagact tgatagccat catcatatcg
     2221 aagtttcact accctttttc catttgccat ctattgaagt aataataggc gcatgcaact
     2281 tcttttcttt ttttttcttt tctctctccc ccgttgttgt ctcaccatat ccgcaatgac
     2341 aaaaaaatga tggaagacac taaaggaaaa aattaacgac aaagacagca ccaacagatg
     2401 tcgttgttcc agagctgatg aggggtatct cgaagcacac gaaacttttt ccttccttca
     2461 ttcacgcaca ctactctcta atgagcaacg gtatacggcc ttccttccag ttacttgaat
     2521 ttgaaataaa aaaaagtttg ctgtcttgct atcaagtata aatagacctg caatccctat
     2581 cagtgataga gatctcccta tcagtgatag agattcttcc ttgtttcttt ttctgcaggc
     2641 tagcatgggt acccgcccat atgcttgccc tgtcgagtcc tgcgatcgcc gcttttctcg
     2701 ctcggatgag cttacccgcc atatccgcat ccataccggt cagaagccct tccagtgtcg
     2761 aatctgcatg cgtaacttca gtcgtagtga ccaccttacc acccacatcc gcacccacac
     2821 aggcgagaag ccttttgcct gtgacatttg tgggaggaag tttgccagga gtgatgaacg
     2881 caagaggcat accaaaatcc atacaggtag atccggtgac ggtgctggtt taattaactc
     2941 tgctggagac atgagagctg ccaacctttg gccaagcccg ctcatgatca aacgctctaa
     3001 gaagaacagc ctggccttgt ccctgacggc cgaccagatg gtcagtgcct tgttggatgc
     3061 tgagcccccc atactctatt ccgagtatga tcctaccaga cccttcagtg aagcttcgat
     3121 gatgggctta ctgaccaacc tggcagacag ggagctggtt cacatgatca actgggcgaa
     3181 gagggtgcca ggctttgtgg atttgaccct ccatgatcag gtccaccttc tagaatgtgc
     3241 ctggctagag atcctgatga ttggtctcgt ctggcgctcc atggagcacc cagggaagct
     3301 actgtttgct cctaacttgc tcttggacag gaaccaggga aaatgtgtag agggcatggt
     3361 ggagatcttc gacatgctgc tggctacatc atctcggttc cgcatgatga atctgcaggg
     3421 agaggagttt gtgtgcctca aatctattat tttgcttaat tctggagtgt acacatttct
     3481 gtccagcacc ctgaagtctc tggaagagaa ggaccatatc caccgagtcc tggacaagat
     3541 cacagacact ttgatccacc tgatggccaa ggcaggcctg accctgcagc agcagcacca
     3601 gcggctggcc cagctcctcc tcatcctctc ccacatcagg cacatgagta acaaaggcat
     3661 ggagcatctg tacagcatga agtgcaagaa cgtggtgccc ctctatgacc tgctgctgga
     3721 gatgctggac gcccaccgcc tacatgcgcc cactagccgt ggaggggcat ccgtggagga
     3781 gacggaccaa agccacttgg ccactgcggg ctctacttca tcgcattcct tgcaaaagta
     3841 ttacatcacg ggggaggcag agggtttccc tgccacagtc gcggccgcag gtgacggtgc
     3901 tggtttaatt aacatgacgg tcgaccatga tttcaatagc gaagatattt tattccccat
     3961 agaaagcatg agtagtatac aatacgtgga gaataataac ccaaataata ttaacaacga
     4021 tgttatcccg tattctctag atatcaaaaa cactgtctta gatagtgcgg atctcaatga
     4081 cattcaaaat caagaaactt cactgaattt ggggcttcct ccactatctt tcgactctcc
     4141 actgcccgta acggaaacga taccatccac taccgataac agcttgcatt tgaaagctga
     4201 tagcaacaaa aatcgcgatg caagaactat tgaaaatgat agtgaaatta agagtactaa
     4261 taatgctagt ggctctgggg caaatcaata cacaactctt acttcacctt atcctatgaa
     4321 cgacattttg tacaacatga acaatccgtt acaatcaccg tcaccttcat cggtacctca
     4381 aaatccgact ataaatcctc ccataaatac agcaagtaac gaaactaatt tatcgcctca
     4441 aacttcaaat ggtaatgaaa ctcttatatc tcctcgagcc caacaacata cgtccattaa
     4501 agataatcgt ctgtccttac ctaatggtgc taattcgaat cttttcattg acactaaccc
     4561 aaacaatttg aacgaaaaac taagaaatca attgaactca gatacaaatt catattctaa
     4621 ctccatttct aattcaaact ccaattctac gggtaattta aattccagtt attttaattc
     4681 actgaacata gactccatgc tagatgatta cgtttctagt gatctcttat tgaatgatga
     4741 tgatgatgac actaatttat cacgccgaag atttagcgac gttataacaa accaatttcc
     4801 gtcaatgaca aattcgagga atgagctcta agcaaatagc taaattatat acgaattaat
     4861 attatgatta agtgtttacg tgagtgcgat atttttatta ctatcttata cagttgtata
     4921 tactctataa aatgagttgt ctattaatta acgcgatgaa tgctttctgg gtttacctct
     4981 ccaacaactc tagtttactt ctcaatacat tcaattgtat ttgatttgtc aatacttcat
     5041 cattaatcaa ttctatagtt ttgtttttct cgtttatttc caaatttaat gcatcaattt
     5101 tattattcaa tttgtcgttg attttggtta atgattttat ggtttgatct ctggcattga
     5161 ttgtttgtgt tagtttttca ttattgataa ttaaattatt taagttagtt atcaactcgg
     5221 tgttttcaag tttcaagttt tcaatttctt tagagtttat tagatttgtc aaagtttctg
     5281 aattgcttga ttggtccggc ccaggtattg ttagcggttt gaagcaggcg gcggaagaag
     5341 taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctag
     5401 ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta
     5461 tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta
     5521 tgacacccgg tgtgggttta gatgacaagg gagacgcatt gggtcaacag tatagaaccg
     5581 tggatgatgt ggtctctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa
     5641 agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg ggtttaaacg
     5701 gccacgctgg cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt
     5761 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     5821 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5881 gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5941 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     6001 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     6061 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     6121 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     6181 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     6241 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     6301 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     6361 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata gggtaataac
     6421 tgatataatt aaattgaagc tctaatttgt gagtttagta ttaggatctc tacccttggc
     6481 gaaaagtcct ctgccaacaa tgatgatatc tgatccacca cttacaactt cgtcgacggt
     6541 tctgtactgc tgacccaata tgcgtatata taccaatcta agtctgtgct ccttccttcg
     6601 ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt caaggaaacc gaaatcaaaa
     6661 aaaagaataa aaaaaaaatg atgaattgaa aaggtggtat ggtgcactct cagtacaatc
     6721 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     6781 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     6841 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     6901 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcggcgcgc
     6961 cgaggacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     7021 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     7081 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     7141 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     7201 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     7261 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     7321 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     7381 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     7441 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     7501 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     7561 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     7621 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     7681 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     7741 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     7801 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     7861 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     7921 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     7981 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     8041 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     8101 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     8161 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     8221 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     8281 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     8341 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     8401 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     8461 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     8521 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     8581 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     8641 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     8701 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     8761 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     8821 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     8881 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     8941 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     9001 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     9061 cggctcctat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     9121 accatgatta cgccaagctc ggaattaacc ctcactaaag ggaacaaaag ctg
//
