LOCUS       sequence-163073-        6921 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cytoplasmic restricted form of N-terminal myc tagged and 
            C-terminal flag tagged mRNA capping enzyme, inactive form.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6921)
  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 6921)
  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: pcDNA4/TO-myc  NES-mCE(K294A) NLS-Flag
FEATURES             Location/Qualifiers
     source          1..6921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1056..1082
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2877..2900
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2929..2946)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2935..3159
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3205..3633
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3292..3311)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3502..3523
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3642..3662)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3647..3976
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3827..3962
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3889..3908
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4024..4071
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4090..4464
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4594..4715
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4631..4650)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4685..4704
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4764..4780)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4764..4780)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4777..4799)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4788..4804
                     /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(4812..4818,4819..4836,4837..4842))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4857..4878
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4995..5012)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5166..5754)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5246..5265)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5925..6716,6717..6785))
                     /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     6548..6567
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6786..6890)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttggtacca ccatggcatc aatgcagaag ctgatctcag
     1021 aggaggacct gcttatggcc atggaggccc ttcagctacc accgcttgag agacttactc
     1081 ttgatatggc catggaggcc gcttacaaca agatcccgcc gcggtggctg aactgtccgc
     1141 ggcgcggtca gccggtggca ggaagattct tacctctgaa gacgatgctg gggccaagat
     1201 atgatagtca agttgctgaa gaaaaccggt tccatcccag tatgctctca aattatctaa
     1261 agagcctgaa ggttaaaatg agcttgttgg tggatctgac aaatacttca aggttctatg
     1321 acagaaatga catagaaaaa gaaggaatca agtatatcaa acttcagtgt aaaggacatg
     1381 gcgaatgccc caccactgag aatactgaga cattcattcg cctgtgtgag aggtttaatg
     1441 aaagaagccc acctgaactt ataggtgttc actgcaccca tggcttcaac cgtactggtt
     1501 tccttatatg tgcctttttg gtggaaaaaa tggattggag tattgaagca gcagttgcta
     1561 catttgccca agccagacca ccaggaatct acaagggtga ttatttgaag gaactttttc
     1621 gtcgctatgg agatatagaa gaagcaccac caccacctgt attgccagat tggtgttttg
     1681 aagatgagga tgaagaagat gaggatgaag atggaaaaaa ggactcagaa ccagggtcaa
     1741 gtgcatcctt tagtaaaagg agaaaagagc ggttaaaact gggagctatt ttcttggaag
     1801 gcataactgt taaaggtgta actcaagtaa caactcaacc caagttagga gaggtgcagc
     1861 agaaatgtca tcagttctgt ggctgggaag ggtctgggtt ccctggagca cagcctgtct
     1921 ccatggacaa gcaaaatatt agacttttag agcagaagcc ttataaagta agctgggcag
     1981 cagacggcac tcgttacatg atgttgattg atggcacaaa tgaagttttt atgattgaca
     2041 gagataattc cgtttttcat gtttcaaatc tggaatttcc atttcgtaaa gatcttcgca
     2101 tgcatttatc aaatactctt ttggatgggg aaatgatcat tgacaaagta aatggacagg
     2161 ccgtcccaag atatttgatc tatgacataa ttaagttcaa tgcacaacca gttggagact
     2221 gtgattttaa tattcgtcta cagtgtattg aacgtgaaat tataagtcca cgacatgaaa
     2281 aaatgaagac tggactcatt gacaaaacac aggaaccatt tagtgtcaga cctaaacaat
     2341 tttttgacat caatatttca agaaagcttc tggaaggaaa ttttgccaaa gaagtcagcc
     2401 atgaaatgga tggacttatt tttcagccta ttggaaaata caagcctggt cgatgtgatg
     2461 acattttgaa atggaaacct cccagtctaa actctgtgga ttttcgactt aagataacaa
     2521 gaatgggagg agaagggttg cttccacaga atgttggcct tctctatgtt ggaggttatg
     2581 aaagaccctt tgcacaaatc aaggtgacaa aagaactaaa acagtatgac aacaaaatta
     2641 tagaatgcaa atttgagaac aatagctggg tcttcatgag acagaggata gacaaaagtt
     2701 tcccaaatgc ctacaacaca gccatggctg tgtgcaatag catctcgaac cctgtcacca
     2761 aggagatgct gtttgaattc attgacagat gtgcagcagc cgcccaggga cagcccctgg
     2821 accctgacac ggagctcatg ccgcccccac cgcccaaaag actgcatcgg ccaaccgatt
     2881 acaaggatga cgacgataag taggggcccg tttaaacccg ctgatcagcc tcgactgtgc
     2941 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     3001 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     3061 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     3121 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     3181 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     3241 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     3301 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3361 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3421 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3481 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3541 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3601 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3661 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3721 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3781 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3841 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3901 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3961 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca
     4021 cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     4081 aactaaacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcgcc
     4141 ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt ggaggacgac
     4201 ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca ggaccaggtg
     4261 gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct gtacgccgag
     4321 tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat gaccgagatc
     4381 ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa ctgcgtgcac
     4441 ttcgtggccg aggagcagga ctgacacgtg ctacgagatt tcgattccac cgccgccttc
     4501 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4561 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4621 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4681 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4741 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4801 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4861 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     4921 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     4981 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5041 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5101 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5161 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5221 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5281 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5341 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5401 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5461 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5521 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5581 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5641 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5701 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5761 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5821 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     5881 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     5941 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6001 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6061 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6121 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6181 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6241 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6301 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6361 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6421 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6481 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6541 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6601 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6661 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6721 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6781 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6841 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6901 cgaaaagtgc cacctgacgt c
//
