LOCUS       sequence-217068-        7406 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression plasmid that can be used with the Flp-In T-REx 
            system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7406)
  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 7406)
  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: pcDNA5/FRT/TO-Intron-CterFlag-MePCE
FEATURES             Location/Qualifiers
     source          1..7406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        93..472
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        473..676
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     627..647
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    678..696
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    699..717
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     727..751
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          870..1002
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     985..1004
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1047..1065
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3147..3170
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(3216..3233)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3222..3446
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3579..3598)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3730..3777
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3785..4807
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3785..4807
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4937..5058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4974..4993)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5028..5047
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5107..5123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5107..5123)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5120..5142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5131..5147
                     /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)."
     promoter        complement(join(5155..5161,5162..5179,5180..5185))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5200..5221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5338..5355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5509..6097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5589..5608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6268..7059,7060..7128))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6891..6910
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7129..7233)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7308..7327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
       61 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
      121 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      181 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      241 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      301 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      361 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      421 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      481 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      541 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      601 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      661 ctatataagc agagctctcc ctatcagtga tagagatctc cctatcagtg atagagatcg
      721 tcgacgagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
      781 cctccataga agacaccggg accgatccag cctccggact ctagcgttta aacttaagct
      841 gcagaagttg gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag
      901 gagaccaata gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac
      961 ctattggtct tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca
     1021 attacagctc ttaaggctag agtacttaat acgactcact ataggctagc atgatcgaga
     1081 tggcggcgga gaaggagccg tttctggtgc cggccccgcc gccgccgctc aaagatgagt
     1141 cgggcggagg gggcggcccc acggtgccac cgcaccaaga ggccgcctct ggggagctcc
     1201 gcggcgggac ggagcgtggt ccgggtcgtt gcgcgccatc tgcggggtcc ccagccgctg
     1261 cggtcggtcg ggaaagcccc ggggccgcgg ccacctcctc cagtggtccc caggcgcagc
     1321 agcaccgagg gggcggcccc caggcgcagt cgcatgggga ggcccgcctg tcggatcccc
     1381 cggggcgagc cgctcccccg gacgtggggg aggagcgccg gggagggggc gggacagagc
     1441 tgggtccccc tgctcctcct cgaccccgca atggctatca gccccaccgg ccacctgggg
     1501 ggggcggggg caagaggaga aatagctgta atgtaggggg aggcggggga ggcttcaaac
     1561 atccggcctt caagaggcgc aggcgggtga attcggactg tgactctgtg ttaccttcca
     1621 acttcctcct ggggggcaat atctttgatc ccctgaacct gaatagcctc ctggatgagg
     1681 aagtgagccg cactctcaac gcggagaccc ctaagtcatc cccccttccg gccaaagggc
     1741 gagatccggt ggagatcctc atccccaaag atattactga cccgctcagt ctcaatactt
     1801 gcactgatga gggccatgta gttcttgctt cgccactcaa gactggtcgg aagcggcata
     1861 gacaccgggg acagcaccac cagcagcagc aggcagccgg agggagtgag agccaccccg
     1921 tgccgcccac agcccctctc acccccttac tccacgggga gggcgcctca cagcagccgc
     1981 ggcacagggg ccagaaccgg gatgcccccc aaccctatga actcaacaca gccatcaact
     2041 gcagggatga agtggtgtct ccccttccat ctgctctgca gggtccctca ggctccctat
     2101 cagcccctcc agctgcctca gttatctctg cacccccatc ttcctcctcc cgacatcgca
     2161 aacgtcgcag gacttccagc aagtcggagg caggggctag gggtggaggc cagggttcca
     2221 aggaaaaggg ccgagggagt tggggaggcc gccaccacca ccaccaccca ctgcctgcag
     2281 caggcttcaa aaagcaacag cgcaagttcc agtatgggaa ttattgcaaa tactatgggt
     2341 accgcaatcc ttcctgtgag gatgggcgcc ttcgggtgtt gaagcctgag tggtttcggg
     2401 gccgggacgt cctagatctg ggctgcaatg tgggccatct gaccctgagc attgcctgca
     2461 agtggggccc gtcccgcatg gtgggcctgg atatcgattc ccggctcatc cattctgccc
     2521 gccaaaacat ccgacactac ctttccgagg agctgcgtct cccaccccag actttggaag
     2581 gggacccggg ggcagagggt gaggaaggga ccaccaccgt tcgaaagagg agctgcttcc
     2641 cagcctcgct gactgccagc cggggtccca tcgctgcccc ccaagtgccc ttggatggag
     2701 cggacacatc agtcttcccc aacaatgttg tcttcgtcac gggtaattat gtgctggatc
     2761 gagatgacct ggtggaggcc caaacacctg agtatgatgt ggtgctctgc ctcagcctca
     2821 ccaagtgggt gcatctgaac tggggagacg agggcctgaa gcgcatgttt cgccggatct
     2881 accggcacct acgccctggg ggcatcctgg tcctagagcc ccaaccctgg tcgtcgtatg
     2941 gcaagagaaa gactcttaca gaaacgatct acaagaacta ctaccgaatc caattgaagc
     3001 cagagcagtt cagttcctac ctgacatccc cagacgtggg cttctccagc tatgagcttg
     3061 tggccacacc ccacaacacc tctaaaggct tccagcgtcc tgtgtacctg ttccacaagg
     3121 cccgatcccc cagccacggc ggccgcgatt acaaggatga cgacgataag tgaggccgct
     3181 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3241 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3301 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3361 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3421 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3481 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3541 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3601 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3661 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3721 acgtacctag aagttcctat tccgaagttc ctattctcta gaaagtatag gaacttcctt
     3781 ggccaaaaag cctgaactca ccgcgacgtc tgtcgagaag tttctgatcg aaaagttcga
     3841 cagcgtctcc gacctgatgc agctctcgga gggcgaagaa tctcgtgctt tcagcttcga
     3901 tgtaggaggg cgtggatatg tcctgcgggt aaatagctgc gccgatggtt tctacaaaga
     3961 tcgttatgtt tatcggcact ttgcatcggc cgcgctcccg attccggaag tgcttgacat
     4021 tggggaattc agcgagagcc tgacctattg catctcccgc cgtgcacagg gtgtcacgtt
     4081 gcaagacctg cctgaaaccg aactgcccgc tgttctgcag ccggtcgcgg aggccatgga
     4141 tgcgatcgct gcggccgatc ttagccagac gagcgggttc ggcccattcg gaccgcaagg
     4201 aatcggtcaa tacactacat ggcgtgattt catatgcgcg attgctgatc cccatgtgta
     4261 tcactggcaa actgtgatgg acgacaccgt cagtgcgtcc gtcgcgcagg ctctcgatga
     4321 gctgatgctt tgggccgagg actgccccga agtccggcac ctcgtgcacg cggatttcgg
     4381 ctccaacaat gtcctgacgg acaatggccg cataacagcg gtcattgact ggagcgaggc
     4441 gatgttcggg gattcccaat acgaggtcgc caacatcttc ttctggaggc cgtggttggc
     4501 ttgtatggag cagcagacgc gctacttcga gcggaggcat ccggagcttg caggatcgcc
     4561 gcggctccgg gcgtatatgc tccgcattgg tcttgaccaa ctctatcaga gcttggttga
     4621 cggcaatttc gatgatgcag cttgggcgca gggtcgatgc gacgcaatcg tccgatccgg
     4681 agccgggact gtcgggcgta cacaaatcgc ccgcagaagc gcggccgtct ggaccgatgg
     4741 ctgtgtagaa gtactcgccg atagtggaaa ccgacgcccc agcactcgtc cgagggcaaa
     4801 ggaatagcac gtactacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     4861 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     4921 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4981 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5041 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5101 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5161 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5221 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5281 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5341 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5401 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5461 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5521 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5581 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5641 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5701 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5761 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5821 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5881 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     5941 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6001 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6061 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6121 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6181 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6241 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6301 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6361 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6421 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6481 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6541 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6601 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6661 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6721 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6781 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6841 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6901 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     6961 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7021 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7081 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7141 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7201 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7261 cgtcgacgga tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct
     7321 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
     7381 tgcgcgagca aaatttaagc tacaac
//
