LOCUS       sequence-217029-        6173 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces an N-terminal GST-tagged MePCE protein consisting of amino 
            acids 1 - 400..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6173)
  AUTHORS   Shelton SB, Shah NM, Abell NS, Devanathan SK, Mercado M, Xhemalce B
  TITLE     Crosstalk between the RNA Methylation and Histone-Binding Activities
            of MePCE Regulates P-TEFb Activation on Chromatin.
  JOURNAL   Cell Rep. 2018 Feb 6;22(6):1374-1383. doi: 
            10.1016/j.celrep.2018.01.028.
  PUBMED    29425494
REFERENCE   2  (bases 1 to 6173)
  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: 
            "10.1016/j.celrep.2018.01"; journalName: "Cell Rep"; date: 
            "2018-02-6- 6"; volume: "22"; issue: "6"; pages: "1374-1383"
COMMENT     Sequence Label: pGEX-5X-1_MePCE_aa1-400
FEATURES             Location/Qualifiers
     source          1..6173
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(105..110,111..126,127..133)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    141..157
                     /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             180..833
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     791..813
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             843..854
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2145..2167)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2312..2330)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2398..2502
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2503..2571,2572..3363)
                     /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(2721..2740)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3534..4122
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4023..4042
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4276..4293
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4366..4443
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4444..5526
                     /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(4463..4482)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5575..5580,5581..5598,5599..5605)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5613..5629
                     /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     5637..5653
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5637..5653
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5649..6173,1..3)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5669..5686)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5669..5685)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5678..5700)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
       61 cgccgacatc ataacggttc tggcaaatat tctgaaatga gctgttgaca attaatcatc
      121 ggctcgtata atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagtattca
      181 tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact cgacttcttt
      241 tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa ggtgataaat
      301 ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat tatattgatg
      361 gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac aagcacaaca
      421 tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga gcggttttgg
      481 atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact ctcaaagttg
      541 attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta tgtcataaaa
      601 catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac gctcttgatg
      661 ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt tgttttaaaa
      721 aacgtattga agctatccca caaattgata agtacttgaa atccagcaag tatatagcat
      781 ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca aaatcggatc
      841 tgatcgaagg tcgtgggatc cccgaattcc cgggtcgact catgatcgag atggcggcgg
      901 agaaggagcc gtttctggtg ccggccccgc cgccgccgct caaagatgag tcgggcggag
      961 ggggcggccc cacggtgcca ccgcaccaag aggccgcctc tggggagctc cgcggcggga
     1021 cggagcgtgg tccgggtcgt tgcgcgccat ctgcggggtc cccagccgct gcggtcggtc
     1081 gggaaagccc cggggccgcg gccacctcct ccagtggtcc ccaggcgcag cagcaccgag
     1141 ggggcggccc ccaggcgcag tcgcatgggg aggcccgcct gtcggatccc ccggggcgag
     1201 ccgctccccc ggacgtgggg gaggagcgcc ggggaggggg cgggacagag ctgggtcccc
     1261 ctgctcctcc tcgaccccgc aatggctatc agccccaccg gccacctggg gggggcgggg
     1321 gcaagaggag aaatagctgt aatgtagggg gaggcggggg aggcttcaaa catccggcct
     1381 tcaagaggcg caggcgggtg aattcggact gtgactctgt gttaccttcc aacttcctcc
     1441 tggggggcaa tatctttgat cccctgaacc tgaatagcct cctggatgag gaagtgagcc
     1501 gcactctcaa cgcggagacc cctaagtcat ccccccttcc ggccaaaggg cgagatccgg
     1561 tggagatcct catccccaaa gatattactg acccgctcag tctcaatact tgcactgatg
     1621 agggccatgt agttcttgct tcgccactca agactggtcg gaagcggcat agacaccggg
     1681 gacagcacca ccagcagcag caggcagccg gagggagtga gagccacccc gtgccgccca
     1741 cagcccctct caccccctta ctccacgggg agggcgcctc acagcagccg cggcacaggg
     1801 gccagaaccg ggatgccccc caaccctatg aactcaacac agccatcaac tgcagggatg
     1861 aagtggtgtc tccccttcca tctgctctgc agggtccctc aggctcccta tcagcccctc
     1921 cagctgcctc agttatctct gcacccccat cttcctcctc ccgacatcgc aaacgtcgca
     1981 ggacttccag caagtcggag gcaggggcta ggggtggagg ccagggttcc aaggaaaagg
     2041 gccgagggag ttggggaggc cgccaccacc accaccaccc atgagcggcc gcatcgtgac
     2101 tgactgacga tctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
     2161 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
     2221 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
     2281 agcggagtgt ataattcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt
     2341 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2401 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2461 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     2521 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     2581 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2641 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2701 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     2761 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2821 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2881 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2941 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3001 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     3061 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3121 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3181 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3241 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3301 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3361 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3421 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3481 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3541 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3601 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3661 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     3721 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3781 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3841 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3901 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3961 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4021 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4081 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4141 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4201 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4261 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4321 tttctcctta cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg
     4381 cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg
     4441 aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc
     4501 gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa
     4561 gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa
     4621 cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt
     4681 gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta
     4741 gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgcgtc
     4801 agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc
     4861 tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt
     4921 attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac
     4981 cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct
     5041 ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac
     5101 tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc
     5161 actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag
     5221 tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc
     5281 tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc
     5341 agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg
     5401 cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc
     5461 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     5521 cagtgagcgc aacgcaatta atgtaagtta gctcactcat taggcacccc aggctttaca
     5581 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     5641 aaacagctat gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg
     5701 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     5761 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     5821 aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg
     5881 atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg
     5941 cgcccatcta caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg
     6001 agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag
     6061 gccagacgcg aattattttt gatggcgttg gaattagctt atcgactgca cggtgcacca
     6121 atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg gctgtgcagg tcg
//
