LOCUS       sequence-217168-        6854 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally bio-myc-tagged mouse CE/Rngtt 211-597 in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6854)
  AUTHORS   Trotman JB, Agana BA, Giltmier AJ, Wysocki VH, Schoenberg DR
  TITLE     RNA-binding proteins and heat-shock protein 90 are constituents of 
            the cytoplasmic capping enzyme interactome.
  JOURNAL   J Biol Chem. 2018 Oct 26;293(43):16596-16607. doi: 
            10.1074/jbc.RA118.004973. Epub 2018 Aug 30.
  PUBMED    30166341
REFERENCE   2  (bases 1 to 6854)
  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: 
            "10.1074/jbc.RA118.004973"; journalName: "J Biol Chem"; date: 
            "2018-10-26- 26"; volume: "293"; issue: "43"; pages: "16596-16607"
COMMENT     Sequence Label: pcDNA3-bio-myc-mCE 211-597
FEATURES             Location/Qualifiers
     source          1..6854
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     655..674
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        655..673
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2199..2217)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2200..2217)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2237..2254)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2243..2467
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2513..2941
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2600..2619)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2810..2831
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2950..2970)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2955..3284
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3135..3270
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3197..3216
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3351..4145
                     /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(3405..3424)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4015..4034
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4319..4440
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4356..4375)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4410..4429
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4489..4505)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4489..4505)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4502..4524)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4513..4529
                     /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(4537..4543,4544..4561,4562..4567))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4582..4603
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4720..4737)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4891..5479)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4971..4990)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5650..6441,6442..6510))
                     /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     6273..6292
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6511..6615)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6690..6709)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgatgtacgg gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac
      661 gactcactat agggagaccc aagcttggca ccatggagtt aaaaaccgcc gcactagctc
      721 aacatgacga ggctgctgga aaggccggtg aaggtgaaat ccctgcccct cttgctggta
      781 ccgtttctaa gatactggta aaagaaggtg acactgttaa agctggtcaa acagttctgg
      841 tgctggaggc tatgaaaatg gagacagaaa ttaacgctcc tactgacgga aaagttgaaa
      901 aggtgttagt taaggaaaga gatgctgttc aaggtggtca aggtctaatc aagatcggcg
      961 ttggcaccat ggcatcaatg cagaagctga tctcagagga ggacctgctt atggccatgg
     1021 aggccgaacc agggtcaagt gcatccttta gtaaaaggag aaaagagcgg ttaaaactgg
     1081 gagctatttt cttggaaggc ataactgtta aaggtgtaac tcaagtaaca actcaaccca
     1141 agttaggaga ggtgcagcag aaatgtcatc agttctgtgg ctgggaaggg tctgggttcc
     1201 ctggagcaca gcctgtctcc atggacaagc aaaatattag acttttagag cagaagcctt
     1261 ataaagtaag ctggaaagca gacggcactc gttacatgat gttgattgat ggcacaaatg
     1321 aagtttttat gattgacaga gataattccg tttttcatgt ttcaaatctg gaatttccat
     1381 ttcgtaaaga tcttcgcatg catttatcaa atactctttt ggatggggaa atgatcattg
     1441 acaaagtaaa tggacaggcc gtcccaagat atttgatcta tgacataatt aagttcaatg
     1501 cacaaccagt tggagactgt gattttaata ttcgtctaca gtgtattgaa cgtgaaatta
     1561 taagtccacg acatgaaaaa atgaagactg gactcattga caaaacacag gaaccattta
     1621 gtgtcagacc taaacaattt tttgacatca atatttcaag aaagcttctg gaaggaaatt
     1681 ttgccaaaga agtcagccat gaaatggatg gacttatttt tcagcctatt ggaaaataca
     1741 agcctggtcg atgtgatgac attttgaaat ggaaacctcc cagtctaaac tctgtggatt
     1801 ttcgacttaa gataacaaga atgggaggag aagggttgct tccacagaat gttggccttc
     1861 tctatgttgg aggttatgaa agaccctttg cacaaatcaa ggtgacaaaa gaactaaaac
     1921 agtatgacaa caaaattata gaatgcaaat ttgagaacaa tagctgggtc ttcatgagac
     1981 agaggataga caaaagtttc ccaaatgcct acaacacagc catggctgtg tgcaatagca
     2041 tctcgaaccc tgtcaccaag gagatgctgt ttgaattcat tgacagatgt gcagcagccg
     2101 cccagggaca gaagcggaag tatcccctgg accctgacac ggagctcatg ccgcccccac
     2161 cgcccaaaag actgcatcgg ccaacctgag ggccctattc tatagtgtca cctaaatgct
     2221 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2281 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2341 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2401 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2461 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     2521 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2581 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2641 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2701 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2761 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2821 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2881 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2941 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3001 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3061 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3121 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3181 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3241 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3301 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3361 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3421 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3481 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     3541 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3601 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3661 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3721 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3781 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3841 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     3901 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3961 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4021 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4081 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4141 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4201 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4261 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     4321 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     4381 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     4441 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     4501 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4561 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4621 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     4681 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     4741 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     4801 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4861 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4921 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4981 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5041 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5101 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5161 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5221 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5281 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5341 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5401 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5461 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5521 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5581 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5641 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5701 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5761 atctggcccc agtgctgcaa tgataccgcg agacccaccc ttggaggctc cagatttatc
     5821 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5881 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5941 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     6001 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     6061 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     6121 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     6181 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     6241 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6301 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6361 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6421 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6481 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6541 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6601 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga
     6661 tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     6721 cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa
     6781 gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt
     6841 tttgcgctgc ttcg
//
