LOCUS       sequence-217030-        5839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces an N-terminal GST-tagged MePCE protein consisting of amino 
            acids 400 - 689..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5839)
  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 5839)
  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_aa400-689
FEATURES             Location/Qualifiers
     source          1..5839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(157..162,163..178,179..185)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    193..209
                     /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             232..885
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     843..865
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             895..906
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1863..1885)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2030..2048)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2116..2220
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2221..2289,2290..3081)
                     /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(2439..2458)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3252..3840
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3741..3760
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3994..4011
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4084..4161
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4162..5244
                     /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(4181..4200)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(5293..5298,5299..5316,5317..5323)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5331..5347
                     /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     5355..5371
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5355..5371
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5387..5404)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5387..5403)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5396..5418)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac
       61 tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca
      121 tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca tcggctcgta
      181 taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagtatt catgtcccct
      241 atactaggtt attggaaaat taagggcctt gtgcaaccca ctcgacttct tttggaatat
      301 cttgaagaaa aatatgaaga gcatttgtat gagcgcgatg aaggtgataa atggcgaaac
      361 aaaaagtttg aattgggttt ggagtttccc aatcttcctt attatattga tggtgatgtt
      421 aaattaacac agtctatggc catcatacgt tatatagctg acaagcacaa catgttgggt
      481 ggttgtccaa aagagcgtgc agagatttca atgcttgaag gagcggtttt ggatattaga
      541 tacggtgttt cgagaattgc atatagtaaa gactttgaaa ctctcaaagt tgattttctt
      601 agcaagctac ctgaaatgct gaaaatgttc gaagatcgtt tatgtcataa aacatattta
      661 aatggtgatc atgtaaccca tcctgacttc atgttgtatg acgctcttga tgttgtttta
      721 tacatggacc caatgtgcct ggatgcgttc ccaaaattag tttgttttaa aaaacgtatt
      781 gaagctatcc cacaaattga taagtacttg aaatccagca agtatatagc atggcctttg
      841 cagggctggc aagccacgtt tggtggtggc gaccatcctc caaaatcgga tctgatcgaa
      901 ggtcgtggga tccccctgcc tgcagcaggc ttcaaaaagc aacagcgcaa gttccagtat
      961 gggaattatt gcaaatacta tgggtaccgc aatccttcct gtgaggatgg gcgccttcgg
     1021 gtgttgaagc ctgagtggtt tcggggccgg gacgtcctag atctgggctg caatgtgggc
     1081 catctgaccc tgagcattgc ctgcaagtgg ggcccgtccc gcatggtggg cctggatatc
     1141 gattcccggc tcatccattc tgcccgccaa aacatccgac actacctttc cgaggagctg
     1201 cgtctcccac cccagacttt ggaaggggac ccgggggcag agggtgagga agggaccacc
     1261 accgttcgaa agaggagctg cttcccagcc tcgctgactg ccagccgggg tcccatcgct
     1321 gccccccaag tgcccttgga tggagcggac acatcagtct tccccaacaa tgttgtcttc
     1381 gtcacgggta attatgtgct ggatcgagat gacctggtgg aggcccaaac acctgagtat
     1441 gatgtggtgc tctgcctcag cctcaccaag tgggtgcatc tgaactgggg agacgagggc
     1501 ctgaagcgca tgtttcgccg gatctaccgg cacctacgcc ctgggggcat cctggtccta
     1561 gagccccaac cctggtcgtc gtatggcaag agaaagactc ttacagaaac gatctacaag
     1621 aactactacc gaatccaatt gaagccagag cagttcagtt cctacctgac atccccagac
     1681 gtgggcttct ccagctatga gcttgtggcc acaccccaca acacctctaa aggcttccag
     1741 cgtcctgtgt acctgttcca caaggcccga tcccccagcc acgaattccc gggtcgactc
     1801 gagcggccgc atcgtgactg actgacgatc tgcctcgcgc gtttcggtga tgacggtgaa
     1861 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     1921 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     1981 acccagtcac gtagcgatag cggagtgtat aattcttgaa gacgaaaggg cctcgtgata
     2041 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     2101 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2161 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2221 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     2281 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2341 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     2401 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2461 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     2521 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2581 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2641 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2701 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2761 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2821 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2881 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     2941 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     3001 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     3061 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     3121 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3181 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3241 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3301 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3361 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     3421 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3481 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3541 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3601 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     3661 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3721 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3781 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3841 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3901 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     3961 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     4021 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcataaat
     4081 tccgacacca tcgaatggcg caaaaccttt cgcggtatgg catgatagcg cccggaagag
     4141 agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc agagtatgcc
     4201 ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt ttctgcgaaa
     4261 acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa ccgcgtggca
     4321 caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag tctggccctg
     4381 cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact gggtgccagc
     4441 gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc ggtgcacaat
     4501 cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga ccaggatgcc
     4561 attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt ctctgaccag
     4621 acacccatca acagtattat tttctcccat gaagacggta cgcgactggg cgtggagcat
     4681 ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag ttctgtctcg
     4741 gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat tcagccgata
     4801 gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat gcaaatgctg
     4861 aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc gctgggcgca
     4921 atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt agtgggatac
     4981 gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa acaggatttt
     5041 cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg ccaggcggtg
     5101 aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct ggcgcccaat
     5161 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     5221 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc tcactcatta
     5281 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     5341 ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg gccgtcgttt
     5401 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcatt gcagcacatc
     5461 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     5521 tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa gcggtgccgg
     5581 aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc tcaaactggc
     5641 agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt acggtcaatc
     5701 cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt aatgttgatg
     5761 aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga attagcttat
     5821 cgactgcacg gtgcaccaa
//
