LOCUS       sequence-163404-        7259 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of myc epitope and murine mRNA Capping Enzyme with 
            Terminal nuclear export signal in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7259)
  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 7259)
  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 NES-mCE
FEATURES             Location/Qualifiers
     source          1..7259
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(40..59)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     650..669
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    954..1075
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(991..1010)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1045..1064
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1124..1140)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1124..1140)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1137..1159)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1148..1164
                     /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(1172..1178,1179..1196,1197..1202))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1217..1238
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1355..1372)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1526..2114)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1606..1625)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2285..3076,3077..3145))
                     /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     2908..2927
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3146..3250)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3325..3344)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3516..3895
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3896..4099
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4050..4070
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4144..4163
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4144..4162
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4248..4274
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     promoter        complement(6093..6111)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6094..6111)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6131..6148)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6137..6361
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6407..6835
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6494..6513)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6704..6725
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6844..6864)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6849..7178
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7029..7164
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7091..7110
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(7245..7259,1..780)
                     /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"
ORIGIN
        1 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
       61 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
      121 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
      181 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
      241 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
      301 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
      361 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
      421 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
      481 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
      541 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
      601 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
      661 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
      721 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
      781 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
      841 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
      901 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
      961 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     1021 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     1081 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     1141 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     1201 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     1261 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     1321 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1381 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1441 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1501 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1561 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1621 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1681 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1741 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1801 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1861 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1921 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     1981 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2041 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2101 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2161 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2221 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2281 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2341 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2401 gccccagtgc tgcaatgata ccgcgagacc cacccttgga ggctccagat ttatcagcaa
     2461 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2521 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2581 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2641 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2701 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2761 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2821 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2881 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2941 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3001 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3061 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3121 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3181 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3241 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     3301 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3361 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     3421 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     3481 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     3541 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     3601 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     3661 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     3721 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     3781 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     3841 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     3901 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     3961 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     4021 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     4081 ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga
     4141 aattaatacg actcactata gggagaccca agcttggtac caccatggca tcaatgcaga
     4201 agctgatctc agaggaggac ctgcttatgg ccatggaggc ccttcagcta ccaccgcttg
     4261 agagacttac tcttgatatg gccatggagg ccgcttacaa caagatcccg ccgcggtggc
     4321 tgaactgtcc gcggcgcggt cagccggtgg caggaagatt cttacctctg aagacgatgc
     4381 tggggccaag atatgatagt caagttgctg aagaaaaccg gttccatccc agtatgctct
     4441 caaattatct aaagagcctg aaggttaaaa tgagcttgtt ggtggatctg acaaatactt
     4501 caaggttcta tgacagaaat gacatagaaa aagaaggaat caagtatatc aaacttcagt
     4561 gtaaaggaca tggcgaatgc cccaccactg agaatactga gacattcatt cgcctgtgtg
     4621 agaggtttaa tgaaagaagc ccacctgaac ttataggtgt tcactgcacc catggcttca
     4681 accgcactgg tttccttata tgtgcctttt tggtggaaaa aatggattgg agtattgaag
     4741 cagcagttgc tacatttgcc caagccagac caccaggaat ctacaagggt gattatttga
     4801 aggaactttt tcgtcgctat ggagatatag aagaagcacc accaccacct gtattgccag
     4861 attggtgttt tgaagatgag gatgaagaag atgaggatga agatggaaaa aaggactcag
     4921 aaccagggtc aagtgcatcc tttagtaaaa ggagaaaaga gcggttaaaa ctgggagcta
     4981 ttttcttgga aggcataact gttaaaggtg taactcaagt aacaactcaa cccaagttag
     5041 gagaggtgca gcagaaatgt catcagttct gtggctggga agggtctggg ttccctggag
     5101 cacagcctgt ctccatggac aagcaaaata ttagactttt agagcagaag ccttataaag
     5161 taagctggaa agcagacggc actcgttaca tgatgttgat tgatggcaca aatgaagttt
     5221 ttatgattga cagagataat tccgtttttc atgtttcaaa tctggaattt ccatttcgta
     5281 aagatcttcg catgcattta tcaaatactc ttttggatgg ggaaatgatc attgacaaag
     5341 taaatggaca ggccgtccca agatatttga tctatgacat aattaagttc aatgcacaac
     5401 cagttggaga ctgtgatttt aatattcgtc tacagtgtat tgaacgtgaa attataagtc
     5461 cacgacatga aaaaatgaag actggactca ttgacaaaac acaggaacca tttagtgtca
     5521 gacctaaaca attttttgac atcaatattt caagaaagct tctggaagga aattttgcca
     5581 aagaagtcag ccatgaaatg gatggactta tttttcagcc tattggaaaa tacaagcctg
     5641 gtcgatgtga tgacattttg aaatggaaac ctcccagtct aaactctgtg gattttcgac
     5701 ttaagataac aagaatggga ggagaagggt tgcttccaca gaatgttggc cttctctatg
     5761 ttggaggtta tgaaagaccc tttgcacaaa tcaaggtgac aaaagaacta aaacagtatg
     5821 acaacaaaat tatagaatgc aaatttgaga acaatagctg ggtcttcatg agacagagga
     5881 tagacaaaag tttcccaaat gcctacaaca cagccatggc tgtgtgcaat agcatctcga
     5941 accctgtcac caaggagatg ctgtttgaat tcattgacag atgtgcagca gccgcccagg
     6001 gacagaagcg gaagtatccc ctggaccctg acacggagct catgccgccc ccaccgccca
     6061 aaagactgca tcggccaacc tgagggccct attctatagt gtcacctaaa tgctagagct
     6121 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     6181 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     6241 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     6301 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     6361 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     6421 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     6481 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     6541 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     6601 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     6661 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     6721 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     6781 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     6841 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     6901 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     6961 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     7021 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     7081 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     7141 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     7201 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagat
//
