LOCUS       sequence-217167-        5130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses N-terminally bio-myc-tagged mouse CE/Rngtt 2-210 in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5130)
  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 5130)
  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 2-210
FEATURES             Location/Qualifiers
     source          1..5130
                     /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(1665..1683)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1666..1683)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1703..1720)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1709..1933
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1979..2407
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2066..2085)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2276..2297
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2416..2436)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2421..2750
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2601..2736
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2663..2682
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2817..3611
                     /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(2871..2890)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3481..3500
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3785..3906
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3822..3841)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3876..3895
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3955..3971)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3955..3971)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3968..3990)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3979..3995
                     /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(4003..4009,4010..4027,4028..4033))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4048..4069
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(4224..4602)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     4549..4568
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4787..4891)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4966..4985)
                     /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 aggccgctta caacaagatc ccgccgcggt ggctgaactg tccgcggcgc ggtcagccgg
     1081 tggcaggaag attcttacct ctgaagacga tgctggggcc aagatatgat agtcaagttg
     1141 ctgaagaaaa ccggttccat cccagtatgc tctcaaatta tctaaagagc ctgaaggtta
     1201 aaatgagctt gttggtggat ctgacaaata cttcaaggtt ctatgacaga aatgacatag
     1261 aaaaagaagg aatcaagtat atcaaacttc agtgtaaagg acatggcgaa tgccccacca
     1321 ctgagaatac tgagacattc attcgcctgt gtgagaggtt taatgaaaga agcccacctg
     1381 aacttatagg tgttcactgc acccatggct tcaaccgcac tggtttcctt atatgtgcct
     1441 ttttggtgga aaaaatggat tggagtattg aagcagcagt tgctacattt gcccaagcca
     1501 gaccaccagg aatctacaag ggtgattatt tgaaggaact ttttcgtcgc tatggagata
     1561 tagaagaagc accaccacca cctgtattgc cagattggtg ttttgaagat gaggatgaag
     1621 aagatgagga tgaagatgga aaaaaggact catgagggcc ctattctata gtgtcaccta
     1681 aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1741 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1801 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1861 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     1921 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     1981 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2041 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2101 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     2161 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     2221 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     2281 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     2341 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2401 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2461 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2521 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2581 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2641 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2701 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2761 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     2821 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2881 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2941 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3001 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     3061 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     3121 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     3181 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     3241 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     3301 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3361 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3421 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3481 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3541 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3601 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3661 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3721 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3781 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3841 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3901 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     3961 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4021 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4081 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4141 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4201 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4261 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4321 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4381 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4441 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4501 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4561 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4621 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4681 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4741 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4801 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4861 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     4921 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     4981 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     5041 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     5101 gcttagggtt aggcgttttg cgctgcttcg
//
