LOCUS       sequence-360055-        6217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mutation Y2404F (TAC/TTC) in GST-TRR-C421 (TRR 2011-2431 aa). 
            Expression in E. coli. by IPTG induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6217)
  AUTHORS   Tie F, Banerjee R, Saiakhova AR, Howard B, Monteith KE, Scacheri PC,
            Cosgrove MS, Harte PJ
  TITLE     Trithorax monomethylates histone H3K4 and interacts directly with 
            CBP to promote H3K27 acetylation and antagonize Polycomb silencing.
  JOURNAL   Development. 2014 Mar;141(5):1129-39. doi: 10.1242/dev.102392.
  PUBMED    24550119
REFERENCE   2  (bases 1 to 6217)
  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"; doi: "10.1242/dev"; 
            journalName: "Development"; date: "2014-03"; volume: "141"; issue: 
            "5"; pages: "1129-39"
COMMENT     Sequence Label: pGEX-2T-TRR-C421-Y2404F
FEATURES             Location/Qualifiers
     source          1..6217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2267..2289)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2434..2452)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2520..2624
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2625..2693,2694..3485)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2843..2862)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3656..4244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4145..4164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4398..4415
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4488..4565
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4566..5648
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4585..4604)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5697..5702,5703..5720,5721..5727)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5735..5751
                     /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     5759..5775
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5759..5775
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5771..6217,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5791..5808)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5791..5807)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5800..5822)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gttccgcgtg gatcccatat gggtgcagga gcaggcgcca
      961 cgactggcgc cggattagga ggctctgtgg cagataatga actgagctcc ctggttgtcc
     1021 accggagagt ttttgtcgat cgggatgaaa accgtctggt ggccaccgtc atgcactatt
     1081 cagagctaag caacctgctc cgtgtgggca acatgacgtt cctgaacgtg ggtcagttac
     1141 tgccgcatca gctggaggcc ttccacacac cccactacat ctaccccatt ggctacaagg
     1201 tgagtcgcta ctactggtgc gtgcgacgac ccaatcgtag gtgtcgatac atttgcagta
     1261 tcgcagaggc cggatgcaag cccgagtttc gcatccaggt gcaggacgcc ggcgacaagg
     1321 agccggagcg cgagtttcgg ggcagttcgc cctcggcggt ctggcagcag attctgcagc
     1381 caatcacgcg actgcgtaag gtgcacaagt ggttgcagct ctttccccag cacatcagtg
     1441 gtgaggacct atttggccta acggaaccgg ccattgtgcg aatcctggaa agcctcccgg
     1501 gcattgagac gctcactgac taccgcttca agtacggtcg caacccgctt cttgagttcc
     1561 cgctagccat caatccgtct ggagcggcgc gtacggaacc caagcaacga cagctgctcg
     1621 tctggaggaa gccgcatacg caacgcacgg ccggcagctg cagcacgcag cggatggcca
     1681 actcggctgc gattgccgga gaagtggcct gcccgtacag caagcagttc gtgcattcga
     1741 agagctccca gtacaaaaag atgaagcagg agtggcgcaa caacgtctac ttggcgagat
     1801 ctaagatcca gggtcttggc ttgtatgcag ctcgggacat tgaaaagcac accatgataa
     1861 tcgagtacat tggcgaggtt atccgcaccg aggtctccga gatacgcgag aagcagtacg
     1921 agtccaagaa ccgtggcatc tacatgttcc ggctggatga agaccgcgtg gtggacgcca
     1981 ctctaagcgg cgggttggcg cgctacatca accattcgtg caatcccaac tgtgttaccg
     2041 agatagtgga ggtcgatcgc gacgtgcgga tcataatatt cgccaagcgg aagatctatc
     2101 gcggcgagga gctctcgttc gactacaagt tcgacattga ggacgagtcg cacaagatac
     2161 cgtgcgcctg tggagcgccc aactgtcgca agtggatgaa catgaatcga attcatcgtg
     2221 actgactgac gatctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
     2281 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
     2341 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
     2401 atagcggagt gtataattct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     2461 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     2521 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2581 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2641 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2701 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2761 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2821 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     2881 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     2941 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3001 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3061 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3121 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     3181 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3241 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3301 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3361 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3421 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3481 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3541 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3601 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3661 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3721 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3781 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3841 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     3901 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     3961 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4021 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     4081 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4141 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4201 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4261 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4321 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4381 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt
     4441 attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac accatcgaat
     4501 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     4561 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     4621 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     4681 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     4741 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     4801 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     4861 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg
     4921 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg
     4981 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta
     5041 ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc
     5101 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg
     5161 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg
     5221 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc
     5281 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg
     5341 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca
     5401 gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa
     5461 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt
     5521 tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc
     5581 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     5641 ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc ccaggcttta
     5701 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     5761 ggaaacagct atgaccatga ttacggattc actggccgtc gttttacaac gtcgtgactg
     5821 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     5881 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     5941 cgaatggcgc tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg
     6001 cgatcttcct gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga
     6061 tgcgcccatc tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac
     6121 ggagaatccg acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga
     6181 aggccagacg cgaattattt ttgatggcgt tggaatt
//
