LOCUS       sequence-163405-        7196 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses myc tag and mRNA capping enzyme without Nuclear 
            Localization Sequence (NLS) from C terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7196)
  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 7196)
  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-mCE NLS
FEATURES             Location/Qualifiers
     source          1..7196
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..56
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             191..985
                     /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(245..264)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     855..874
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1159..1280
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1196..1215)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1250..1269
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1329..1345)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1329..1345)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1342..1364)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1353..1369
                     /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(1377..1383,1384..1401,1402..1407))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1422..1443
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1560..1577)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1731..2319)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1811..1830)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2490..3281,3282..3350))
                     /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     3113..3132
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3351..3455)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3530..3549)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3721..4100
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4101..4304
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4255..4275
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4349..4368
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4349..4367
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(6235..6253)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6236..6253)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6273..6290)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6279..6503
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6549..6977
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6636..6655)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6846..6867
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6986..7006)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        join(6991..7196,1..124)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7171..7196,1..110)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
       61 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      121 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
      181 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
      241 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
      301 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
      361 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
      421 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
      481 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
      541 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
      601 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
      661 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
      721 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
      781 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
      841 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
      901 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
      961 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     1021 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     1081 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     1141 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1201 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1261 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     1321 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     1381 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     1441 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1501 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1561 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1621 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1681 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1741 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1801 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1861 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1921 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1981 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2041 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2101 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2161 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2221 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2281 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2341 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2401 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2461 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2521 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2581 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccaccc
     2641 ttggaggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2701 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2761 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2821 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2881 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2941 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3001 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3061 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3121 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3181 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3241 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3301 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3361 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3421 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3481 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     3541 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     3601 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     3661 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     3721 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     3781 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     3841 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     3901 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     3961 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4021 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4081 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
     4141 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     4201 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
     4261 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
     4321 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
     4381 ggtaccacca tggcatcaat gcagaagctg atctcagagg aggacctgct tatggccatg
     4441 gaggccgctt acaacaagat cccgccgcgg tggctgaact gtccgcggcg cggtcagccg
     4501 gtggcaggaa gattcttacc tctgaagacg atgctggggc caagatatga tagtcaagtt
     4561 gctgaagaaa accggttcca tcccagtatg ctctcaaatt atctaaagag cctgaaggtt
     4621 aaaatgagct tgttggtgga tctgacaaat acttcaaggt tctatgacag aaatgacata
     4681 gaaaaagaag gaatcaagta tatcaaactt cagtgtaaag gacatggcga atgccccacc
     4741 actgagaata ctgagacatt cattcgcctg tgtgagaggt ttaatgaaag aagcccacct
     4801 gaacttatag gtgttcactg cacccatggc ttcaaccgca ctggtttcct tatatgtgcc
     4861 tttttggtgg aaaaaatgga ttggagtatt gaagcagcag ttgctacatt tgcccaagcc
     4921 agaccaccag gaatctacaa gggtgattat ttgaaggaac tttttcgtcg ctatggagat
     4981 atagaagaag caccaccacc acctgtattg ccagattggt gttttgaaga tgaggatgaa
     5041 gaagatgagg atgaagatgg aaaaaaggac tcagaaccag ggtcaagtgc atcctttagt
     5101 aaaaggagaa aagagcggtt aaaactggga gctattttct tggaaggcat aactgttaaa
     5161 ggtgtaactc aagtaacaac tcaacccaag ttaggagagg tgcagcagaa atgtcatcag
     5221 ttctgtggct gggaagggtc tgggttccct ggagcacagc ctgtctccat ggacaagcaa
     5281 aatattagac ttttagagca gaagccttat aaagtaagct ggaaagcaga cggcactcgt
     5341 tacatgatgt tgattgatgg cacaaatgaa gtttttatga ttgacagaga taattccgtt
     5401 tttcatgttt caaatctgga atttccattt cgtaaagatc ttcgcatgca tttatcaaat
     5461 actcttttgg atggggaaat gatcattgac aaagtaaatg gacaggccgt cccaagatat
     5521 ttgatctatg acataattaa gttcaatgca caaccagttg gagactgtga ttttaatatt
     5581 cgtctacagt gtattgaacg tgaaattata agtccacgac atgaaaaaat gaagactgga
     5641 ctcattgaca aaacacagga accatttagt gtcagaccta aacaattttt tgacatcaat
     5701 atttcaagaa agcttctgga aggaaatttt gccaaagaag tcagccatga aatggatgga
     5761 cttatttttc agcctattgg aaaatacaag cctggtcgat gtgatgacat tttgaaatgg
     5821 aaacctccca gtctaaactc tgtggatttt cgacttaaga taacaagaat gggaggagaa
     5881 gggttgcttc cacagaatgt tggccttctc tatgttggag gttatgaaag accctttgca
     5941 caaatcaagg tgacaaaaga actaaaacag tatgacaaca aaattataga atgcaaattt
     6001 gagaacaata gctgggtctt catgagacag aggatagaca aaagtttccc aaatgcctac
     6061 aacacagcca tggctgtgtg caatagcatc tcgaaccctg tcaccaagga gatgctgttt
     6121 gaattcattg acagatgtgc agcagccgcc cagggacagc ccctggaccc tgacacggag
     6181 ctcatgccgc ccccaccgcc caaaagactg catcggccaa cctgagggcc ctattctata
     6241 gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     6301 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     6361 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     6421 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     6481 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     6541 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     6601 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     6661 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     6721 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     6781 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     6841 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     6901 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     6961 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     7021 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     7081 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     7141 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttc
//
