LOCUS       sequence-360053-        7209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mutation E3616S (GAA/TCA), PCR product coding C-terminal residues 
            2976-3726 of TRX was inserted into modified pGEX-2T by Nde I-Nsi I 
            sites. Expression in E. coli. by IPTG induction.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7209)
  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 7209)
  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-TRX-C751-E3616S
FEATURES             Location/Qualifiers
     source          1..7209
                     /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(3259..3281)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3426..3444)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3512..3616
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3617..3685,3686..4477)
                     /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(3835..3854)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4648..5236
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5137..5156
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5390..5407
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        5480..5557
                     /gene="lacI"
                     /label=lacI promoter
     CDS             5558..6640
                     /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(5577..5596)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(6689..6694,6695..6712,6713..6719)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6727..6743
                     /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     6751..6767
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6751..6767
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(6763..7209,1..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(6783..6800)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6783..6799)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6792..6814)
                     /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 gatccatatt ccccagcagc
      961 aacaacctct gcagcaacaa caggtgcagg tacagccgtc aatgcccatt ataacacttg
     1021 ctgaggcacc agtagtgcaa tcgcaatttg tgatggaacc tcaagctttg gagcagcagg
     1081 agctggctaa ccgcgtgcag catttctcca caagcagcag cagtagcagt agcaactgct
     1141 cactgcccac caatgtggtt aatcctatgc aacaacaagc accatcaacc accagcagct
     1201 ccacaaccag gcctacaaat cgggtgctac cgatgcagca gcgccaggaa cctgctccgt
     1261 tgtccaacga gtgtcccgta gtttcgtcac ccactcctcc gaaacctgtt gaacagccta
     1321 taatccatca aatgactagc gccagtgtgt ccaagtgcta tgcccaaaag tcaacactac
     1381 catcaccagt ttacgaggcg gaactcaaag ttagttcagt tttggagagc attgtgccag
     1441 atgtcacaat ggatgcaatc ttggaggaac agccggtaac ggaatcaatc tatacggagg
     1501 gactctacga gaagaattcg ccgggggagt cgaagactga acagctactt ctccaacaac
     1561 agcagcgaga gcaacttaat caacaattgg tcaacaacgg gtatctgctc gataagcaca
     1621 cctttcaagt ggagccaatg gatacggatg tctacaggga ggaagacctc gaagaggagg
     1681 aggacgagga tgatgatttc agcttgaaaa tggccacatc ggcgtgcaac gatcacgaga
     1741 tgtccgacag tgaagaaccg gcggtcaagg ataagatcag taagattctg gacaacctaa
     1801 ccaacgatga ctgtgcagac tcaatagcca cggcgacgac gatggaagtg gatgccagcg
     1861 ctggttacca gcaaatggtg gaggatgtcc tagccaccac tgcagcacaa tctgctccca
     1921 ccgaagagtt cgaaggcgct ctagaaactg cagctgtgga agccgctgcc acctatatca
     1981 acgagatggc cgatgctcat gtactggacc ttaagcagct gcaaaatgga gtggaactag
     2041 agctcaggag gagaaaagaa gagcaacgaa cggtctcgca agaacaggag caatcaaagg
     2101 ccgccatcgt gcctacagca gctgctcccg aaccaccgca acccattcaa gagcccaaaa
     2161 aaatgactgg tccacatctg ctttatgaaa tacaaagcga agatggcttt acgtacaaat
     2221 cgagttccat aactgagatt tgggagaagg tctttgaggc agtacaggtg gccaggcggg
     2281 cacatggact tactccgctg ccagagggtc ctctagccga tatgggtggc atccagatga
     2341 taggtcttaa aacgaatgcc cttaagtacc taattgaaca gttgccaggc gtggaaaagt
     2401 gctcaaaata tactcccaag tatcacaagc gcaatggtaa cgtgtccaca gcggcaaatg
     2461 gagcacatgg aggcaactta ggcggaagca gtgcatccgc tgccctctca gtgtctggtg
     2521 gagattcaca tggtctgctc gactatggat cggatcagga tgaattggag gaaaatgctt
     2581 acgattgcgc ccgatgtgag ccttattcca accgcagcga gtatgatatg tttagttggt
     2641 tggcctccag acatcgcaag cagcccatcc aggtgttcgt ccagccgtcc gacaatgaac
     2701 tggtacccag gaggggaact ggtagcaatc tacccatggc catgaagtat cggactctta
     2761 aggaaactta caaggattat gttggagtct tcagatccca tatccatgga cgaggcctct
     2821 actgcactaa ggacattgaa gcgggtgaaa tggttatctc atatgccggt gagttgattc
     2881 gctctacact gacggacaaa agggaacgct actacgacag ccgcggcatt gggtgctata
     2941 tgttcaaaat tgacgataac cttgtagttg atgccacgat gcgcggcaat gcggcgcgtt
     3001 tcattaatca ctgctgtgag ccgaattgct attcgaaagt ggtggacatc ttagggcaca
     3061 agcacatcat catctttgcg ctgcgccgaa tcgtgcaggg cgaggagctt acctacgact
     3121 acaaattccc attcgaggat gagaagatac cttgctcatg cggttcgaaa aggtgtcgga
     3181 agtacttaaa ttaaatgcat gaattcatcg tgactgactg acgatctgcc tcgcgcgttt
     3241 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3301 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     3361 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtataatt cttgaagacg
     3421 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     3481 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3541 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3601 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     3661 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     3721 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     3781 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     3841 tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     3901 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     3961 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     4021 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     4081 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     4141 gcgtgacacc acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga
     4201 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     4261 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     4321 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     4381 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     4441 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     4501 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     4561 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     4621 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     4681 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     4741 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4801 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4861 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4921 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4981 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     5041 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     5101 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     5161 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     5221 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     5281 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     5341 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     5401 gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     5461 ttcacaccgc ataaattccg acaccatcga atggcgcaaa acctttcgcg gtatggcatg
     5521 atagcgcccg gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga
     5581 tgtcgcagag tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag
     5641 ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat
     5701 tcccaaccgc gtggcacaac aactggcggg caaacagtcg ttgctgattg gcgttgccac
     5761 ctccagtctg gccctgcacg cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga
     5821 tcaactgggt gccagcgtgg tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa
     5881 agcggcggtg cacaatcttc tcgcgcaacg cgtcagtggg ctgatcatta actatccgct
     5941 ggatgaccag gatgccattg ctgtggaagc tgcctgcact aatgttccgg cgttatttct
     6001 tgatgtctct gaccagacac ccatcaacag tattattttc tcccatgaag acggtacgcg
     6061 actgggcgtg gagcatctgg tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc
     6121 attaagttct gtctcggcgc gtctgcgtct ggctggctgg cataaatatc tcactcgcaa
     6181 tcaaattcag ccgatagcgg aacgggaagg cgactggagt gccatgtccg gttttcaaca
     6241 aaccatgcaa atgctgaatg agggcatcgt tcccactgcg atgctggttg ccaacgatca
     6301 gatggcgctg ggcgcaatgc gcgccattac cgagtccggg ctgcgcgttg gtgcggatat
     6361 ctcggtagtg ggatacgacg ataccgaaga cagctcatgt tatatcccgc cgttaaccac
     6421 catcaaacag gattttcgcc tgctggggca aaccagcgtg gaccgcttgc tgcaactctc
     6481 tcagggccag gcggtgaagg gcaatcagct gttgcccgtc tcactggtga aaagaaaaac
     6541 caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6601 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtaa
     6661 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     6721 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat
     6781 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     6841 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     6901 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gctttgcctg gtttccggca
     6961 ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga tactgtcgtc
     7021 gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa cgtgacctat
     7081 cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg ttactcgctc
     7141 acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat ttttgatggc
     7201 gttggaatt
//
