LOCUS       sequence-162884-        7247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cytoplasmic restricted form of myc tagged mRNA capping 
            enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7247)
  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 7247)
  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 NLS
FEATURES             Location/Qualifiers
     source          1..7247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     570..589
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    874..995
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(911..930)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     965..984
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1044..1060)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1044..1060)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1057..1079)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1068..1084
                     /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(1092..1098,1099..1116,1117..1122))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1137..1158
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1275..1292)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1446..2034)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1526..1545)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2205..2996,2997..3065))
                     /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     2828..2847
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3066..3170)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3245..3264)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3436..3815
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3816..4019
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3970..3990
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4064..4083
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4064..4082
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4168..4194
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     promoter        complement(6001..6019)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6002..6019)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6039..6056)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6045..6269
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6315..6743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6402..6421)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6612..6633
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6752..6772)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6757..7086
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6937..7072
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6999..7018
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             join(7153..7247,1..700)
                     /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(7207..7226)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
       61 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
      121 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
      181 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
      241 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
      301 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
      361 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
      421 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
      481 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
      541 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
      601 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
      661 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
      721 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
      781 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
      841 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
      901 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
      961 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     1021 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1081 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1141 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1201 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1261 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1321 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1381 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1441 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1501 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1561 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1621 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1681 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1741 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1801 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1861 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     1921 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1981 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2041 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2101 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2161 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2221 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2281 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2341 ccgcgagacc cacccttgga ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2401 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2461 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2521 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2581 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2641 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2701 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2761 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2821 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     2881 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     2941 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3001 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3061 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3121 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3181 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     3241 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     3301 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
     3361 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     3421 ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca attacggggt
     3481 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     3541 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     3601 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     3661 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     3721 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     3781 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
     3841 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     3901 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
     3961 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     4021 tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg actcactata
     4081 gggagaccca agcttggtac caccatggca tcaatgcaga agctgatctc agaggaggac
     4141 ctgcttatgg ccatggaggc ccttcagcta ccaccgcttg agagacttac tcttgatatg
     4201 gccatggagg ccgcttacaa caagatcccg ccgcggtggc tgaactgtcc gcggcgcggt
     4261 cagccggtgg caggaagatt cttacctctg aagacgatgc tggggccaag atatgatagt
     4321 caagttgctg aagaaaaccg gttccatccc agtatgctct caaattatct aaagagcctg
     4381 aaggttaaaa tgagcttgtt ggtggatctg acaaatactt caaggttcta tgacagaaat
     4441 gacatagaaa aagaaggaat caagtatatc aaacttcagt gtaaaggaca tggcgaatgc
     4501 cccaccactg agaatactga gacattcatt cgcctgtgtg agaggtttaa tgaaagaagc
     4561 ccacctgaac ttataggtgt tcactgcacc catggcttca accgcactgg tttccttata
     4621 tgtgcctttt tggtggaaaa aatggattgg agtattgaag cagcagttgc tacatttgcc
     4681 caagccagac caccaggaat ctacaagggt gattatttga aggaactttt tcgtcgctat
     4741 ggagatatag aagaagcacc accaccacct gtattgccag attggtgttt tgaagatgag
     4801 gatgaagaag atgaggatga agatggaaaa aaggactcag aaccagggtc aagtgcatcc
     4861 tttagtaaaa ggagaaaaga gcggttaaaa ctgggagcta ttttcttgga aggcataact
     4921 gttaaaggtg taactcaagt aacaactcaa cccaagttag gagaggtgca gcagaaatgt
     4981 catcagttct gtggctggga agggtctggg ttccctggag cacagcctgt ctccatggac
     5041 aagcaaaata ttagactttt agagcagaag ccttataaag taagctggaa agcagacggc
     5101 actcgttaca tgatgttgat tgatggcaca aatgaagttt ttatgattga cagagataat
     5161 tccgtttttc atgtttcaaa tctggaattt ccatttcgta aagatcttcg catgcattta
     5221 tcaaatactc ttttggatgg ggaaatgatc attgacaaag taaatggaca ggccgtccca
     5281 agatatttga tctatgacat aattaagttc aatgcacaac cagttggaga ctgtgatttt
     5341 aatattcgtc tacagtgtat tgaacgtgaa attataagtc cacgacatga aaaaatgaag
     5401 actggactca ttgacaaaac acaggaacca tttagtgtca gacctaaaca attttttgac
     5461 atcaatattt caagaaagct tctggaagga aattttgcca aagaagtcag ccatgaaatg
     5521 gatggactta tttttcagcc tattggaaaa tacaagcctg gtcgatgtga tgacattttg
     5581 aaatggaaac ctcccagtct aaactctgtg gattttcgac ttaagataac aagaatggga
     5641 ggagaagggt tgcttccaca gaatgttggc cttctctatg ttggaggtta tgaaagaccc
     5701 tttgcacaaa tcaaggtgac aaaagaacta aaacagtatg acaacaaaat tatagaatgc
     5761 aaatttgaga acaatagctg ggtcttcatg agacagagga tagacaaaag tttcccaaat
     5821 gcctacaaca cagccatggc tgtgtgcaat agcatctcga accctgtcac caaggagatg
     5881 ctgtttgaat tcattgacag atgtgcagca gccgcccagg gacagcccct ggaccctgac
     5941 acggagctca tgccgccccc accgcccaaa agactgcatc ggccaacctg agggccctat
     6001 tctatagtgt cacctaaatg ctagagctcg ctgatcagcc tcgactgtgc cttctagttg
     6061 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     6121 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     6181 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     6241 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     6301 tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     6361 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     6421 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     6481 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6541 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6601 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     6661 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     6721 ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg gtgtggaaag
     6781 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     6841 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     6901 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6961 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     7021 gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     7081 tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga gacaggatga
     7141 ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg
     7201 gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctc
//
