LOCUS       sequence-163408-        7208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of myc epitope and catalytic inactive form of murine mRNA
            Capping Enzyme (K294A) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7208)
  AUTHORS   Otsuka Y, Kedersha NL, Schoenberg DR
  TITLE     Identification of a cytoplasmic complex that adds a cap onto 
            5'-monophosphate RNA.
  JOURNAL   Mol Cell Biol. 2009 Apr;29(8):2155-67. doi: 10.1128/MCB.01325-08. 
            Epub 2009 Feb 17.
  PUBMED    19223470
REFERENCE   2  (bases 1 to 7208)
  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: 
            "10.1128/MCB.01325-08"; journalName: "Mol Cell Biol"; date: 
            "2009-04"; volume: "29"; issue: "8"; pages: "2155-67"
COMMENT     Sequence Label: pcDNA3-myc-mCE(K294A)
FEATURES             Location/Qualifiers
     source          1..7208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     32..51
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             186..980
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(240..259)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     850..869
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1154..1275
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1191..1210)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1245..1264
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1324..1340)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1324..1340)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1337..1359)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1348..1364
                     /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(1372..1378,1379..1396,1397..1402))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1417..1438
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1555..1572)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1726..2314)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1806..1825)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2485..3276,3277..3345))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGASKGGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3108..3127
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3346..3450)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3525..3544)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3716..4095
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4096..4299
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4250..4270
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4344..4363
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4344..4362
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6242..6260)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6243..6260)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6280..6297)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6286..6510
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6556..6984
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6643..6662)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6853..6874
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6993..7013)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6998..7208,1..119)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7178..7208,1..105)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
       61 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      121 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
      181 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
      241 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
      301 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
      361 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
      421 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
      481 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
      541 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
      601 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
      661 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
      721 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
      781 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
      841 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
      901 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
      961 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     1021 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     1081 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     1141 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1201 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1261 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     1321 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     1381 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     1441 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1501 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1561 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1621 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1681 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     1741 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     1801 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     1861 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     1921 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     1981 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2041 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2101 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2161 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2221 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2281 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2341 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2401 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2461 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2521 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2581 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacccttgga
     2641 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     2701 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     2761 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     2821 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     2881 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     2941 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3001 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3061 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3121 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3181 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3241 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3301 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3361 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3421 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     3481 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     3541 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     3601 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     3661 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
     3721 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     3781 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     3841 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     3901 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     3961 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     4021 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     4081 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     4141 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     4201 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     4261 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
     4321 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agcttggtac
     4381 caccatggca tcaatgcaga agctgatctc agaggaggac ctgcttatgg ccatggaggc
     4441 cgcttacaac aagatcccgc cgcggtggct gaactgtccg cggcgcggtc agccggtggc
     4501 aggaagattc ttacctctga agacgatgct ggggccaaga tatgatagtc aagttgctga
     4561 agaaaaccgg ttccatccca gtatgctctc aaattatcta aagagcctga aggttaaaat
     4621 gagcttgttg gtggatctga caaatacttc aaggttctat gacagaaatg acatagaaaa
     4681 agaaggaatc aagtatatca aacttcagtg taaaggacat ggcgaatgcc ccaccactga
     4741 gaatactgag acattcattc gcctgtgtga gaggtttaat gaaagaagcc cacctgaact
     4801 tataggtgtt cactgcaccc atggcttcaa ccgtactggt ttccttatat gtgccttttt
     4861 ggtggaaaaa atggattgga gtattgaagc agcagttgct acatttgccc aagccagacc
     4921 accaggaatc tacaagggtg attatttgaa ggaacttttt cgtcgctatg gagatataga
     4981 agaagcacca ccaccacctg tattgccaga ttggtgtttt gaagatgagg atgaagaaga
     5041 tgaggatgaa gatggaaaaa aggactcaga accagggtca agtgcatcct ttagtaaaag
     5101 gagaaaagag cggttaaaac tgggagctat tttcttggaa ggcataactg ttaaaggtgt
     5161 aactcaagta acaactcaac ccaagttagg agaggtgcag cagaaatgtc atcagttctg
     5221 tggctgggaa gggtctgggt tccctggagc acagcctgtc tccatggaca agcaaaatat
     5281 tagactttta gagcagaagc cttataaagt aagctgggca gcagacggca ctcgttacat
     5341 gatgttgatt gatggcacaa atgaagtttt tatgattgac agagataatt ccgtttttca
     5401 tgtttcaaat ctggaatttc catttcgtaa agatcttcgc atgcatttat caaatactct
     5461 tttggatggg gaaatgatca ttgacaaagt aaatggacag gccgtcccaa gatatttgat
     5521 ctatgacata attaagttca atgcacaacc agttggagac tgtgatttta atattcgtct
     5581 acagtgtatt gaacgtgaaa ttataagtcc acgacatgaa aaaatgaaga ctggactcat
     5641 tgacaaaaca caggaaccat ttagtgtcag acctaaacaa ttttttgaca tcaatatttc
     5701 aagaaagctt ctggaaggaa attttgccaa agaagtcagc catgaaatgg atggacttat
     5761 ttttcagcct attggaaaat acaagcctgg tcgatgtgat gacattttga aatggaaacc
     5821 tcccagtcta aactctgtgg attttcgact taagataaca agaatgggag gagaagggtt
     5881 gcttccacag aatgttggcc ttctctatgt tggaggttat gaaagaccct ttgcacaaat
     5941 caaggtgaca aaagaactaa aacagtatga caacaaaatt atagaatgca aatttgagaa
     6001 caatagctgg gtcttcatga gacagaggat agacaaaagt ttcccaaatg cctacaacac
     6061 agccatggct gtgtgcaata gcatctcgaa ccctgtcacc aaggagatgc tgtttgaatt
     6121 cattgacaga tgtgcagcag ccgcccaggg acagaagcgg aagtatcccc tggaccctga
     6181 cacggagctc atgccgcccc caccgcccaa aagactgcat cggccaacct gagggcccta
     6241 ttctatagtg tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt
     6301 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     6361 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     6421 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
     6481 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
     6541 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     6601 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     6661 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     6721 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     6781 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     6841 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     6901 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6961 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     7021 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     7081 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     7141 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     7201 ttccgccc
//
