LOCUS       sequence-217048-        5629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Produces an N-terminal GST-tagged MePCE protein consisting of amino 
            acids 470 - 689..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5629)
  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 5629)
  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_aa470-689
FEATURES             Location/Qualifiers
     source          1..5629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(9..14,15..30,31..37)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    45..61
                     /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             84..737
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     695..717
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             747..758
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(1505..1527)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1672..1690)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1758..1862
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1863..1931,1932..2723)
                     /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(2081..2100)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2894..3482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3383..3402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3636..3653
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        3726..3803
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3804..4886
                     /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(3823..3842)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(4935..4940,4941..4958,4959..4965)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4973..4989
                     /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     4997..5013
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4997..5013
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             5009..5536
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5029..5046)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5029..5045)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5038..5060)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 atgagctgtt gacaattaat catcggctcg tataatgtgt ggaattgtga gcggataaca
       61 atttcacaca ggaaacagta ttcatgtccc ctatactagg ttattggaaa attaagggcc
      121 ttgtgcaacc cactcgactt cttttggaat atcttgaaga aaaatatgaa gagcatttgt
      181 atgagcgcga tgaaggtgat aaatggcgaa acaaaaagtt tgaattgggt ttggagtttc
      241 ccaatcttcc ttattatatt gatggtgatg ttaaattaac acagtctatg gccatcatac
      301 gttatatagc tgacaagcac aacatgttgg gtggttgtcc aaaagagcgt gcagagattt
      361 caatgcttga aggagcggtt ttggatatta gatacggtgt ttcgagaatt gcatatagta
      421 aagactttga aactctcaaa gttgattttc ttagcaagct acctgaaatg ctgaaaatgt
      481 tcgaagatcg tttatgtcat aaaacatatt taaatggtga tcatgtaacc catcctgact
      541 tcatgttgta tgacgctctt gatgttgttt tatacatgga cccaatgtgc ctggatgcgt
      601 tcccaaaatt agtttgtttt aaaaaacgta ttgaagctat cccacaaatt gataagtact
      661 tgaaatccag caagtatata gcatggcctt tgcagggctg gcaagccacg tttggtggtg
      721 gcgaccatcc tccaaaatcg gatctgatcg aaggtcgtgg gatccccgtg ggcctggata
      781 tcgattcccg gctcatccat tctgcccgcc aaaacatccg acactacctt tccgaggagc
      841 tgcgtctccc accccagact ttggaagggg acccgggggc agagggtgag gaagggacca
      901 ccaccgttcg aaagaggagc tgcttcccag cctcgctgac tgccagccgg ggtcccatcg
      961 ctgcccccca agtgcccttg gatggagcgg acacatcagt cttccccaac aatgttgtct
     1021 tcgtcacggg taattatgtg ctggatcgag atgacctggt ggaggcccaa acacctgagt
     1081 atgatgtggt gctctgcctc agcctcacca agtgggtgca tctgaactgg ggagacgagg
     1141 gcctgaagcg catgtttcgc cggatctacc ggcacctacg ccctgggggc atcctggtcc
     1201 tagagcccca accctggtcg tcgtatggca agagaaagac tcttacagaa acgatctaca
     1261 agaactacta ccgaatccaa ttgaagccag agcagttcag ttcctacctg acatccccag
     1321 acgtgggctt ctccagctat gagcttgtgg ccacacccca caacacctct aaaggcttcc
     1381 agcgtcctgt gtacctgttc cacaaggccc gatcccccag ccacgaattc ccgggtcgac
     1441 tcgagcggcc gcatcgtgac tgactgacga tctgcctcgc gcgtttcggt gatgacggtg
     1501 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     1561 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca
     1621 tgacccagtc acgtagcgat agcggagtgt ataattcttg aagacgaaag ggcctcgtga
     1681 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     1741 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1801 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1861 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1921 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1981 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     2041 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     2101 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2161 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2221 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2281 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2341 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2401 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2461 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2521 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2581 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2641 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2701 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2761 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2821 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2881 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2941 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     3001 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     3061 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3121 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3181 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3241 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3301 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3361 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3421 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3481 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3541 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     3601 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     3661 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa
     3721 attccgacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
     3781 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
     3841 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
     3901 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
     3961 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
     4021 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
     4081 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
     4141 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
     4201 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
     4261 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
     4321 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
     4381 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
     4441 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     4501 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     4561 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     4621 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     4681 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     4741 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     4801 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     4861 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
     4921 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     4981 ggataacaat ttcacacagg aaacagctat gaccatgatt acggattcac tggccgtcgt
     5041 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     5101 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     5161 gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc
     5221 ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg
     5281 gcagatgcac ggttacgatg cgcccatcta caccaacgtg acctatccca ttacggtcaa
     5341 tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga
     5401 tgaaagctgg ctacaggaag gccagacgcg aattattttt gatggcgttg gaattagctt
     5461 atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg
     5521 gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     5581 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaa
//
