LOCUS       sequence-163407-        7208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of myc epitope and murine mRNA Capping Enzyme in 
            mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7208)
  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 7208)
  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
FEATURES             Location/Qualifiers
     source          1..7208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     139..158
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    291..515
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     504..521
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        541..559
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     541..558
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2438..2457)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2439..2457)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2502..2705)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2531..2551)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2706..3085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     3257..3276
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3351..3455
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3456..3524,3525..4316)
                     /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     complement(3674..3693)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4487..5075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4976..4995
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5229..5246
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5363..5384
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5399..5404,5405..5422,5423..5429)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5437..5453
                     /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)."
     primer_bind     5442..5464
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5461..5477
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5461..5477
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    5526..5647
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5537..5556)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5591..5610
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5821..6615)
                     /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(5932..5951)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     6542..6561
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(6682..7011)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6696..6831
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6750..6769)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6996..7016
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(join(7025..7208,1..245))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7135..7156)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
       61 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
      121 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
      181 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
      241 cgcgtgggga taccccctag agccccagct ggttctttcc gcctcagaag ccatagagcc
      301 caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc
      361 caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt
      421 ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg
      481 gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgatcagc gagctctagc
      541 atttaggtga cactatagaa tagggccctc aggttggccg atgcagtctt ttgggcggtg
      601 ggggcggcat gagctccgtg tcagggtcca ggggatactt ccgcttctgt ccctgggcgg
      661 ctgctgcaca tctgtcaatg aattcaaaca gcatctcctt ggtgacaggg ttcgagatgc
      721 tattgcacac agccatggct gtgttgtagg catttgggaa acttttgtct atcctctgtc
      781 tcatgaagac ccagctattg ttctcaaatt tgcattctat aattttgttg tcatactgtt
      841 ttagttcttt tgtcaccttg atttgtgcaa agggtctttc ataacctcca acatagagaa
      901 ggccaacatt ctgtggaagc aacccttctc ctcccattct tgttatctta agtcgaaaat
      961 ccacagagtt tagactggga ggtttccatt tcaaaatgtc atcacatcga ccaggcttgt
     1021 attttccaat aggctgaaaa ataagtccat ccatttcatg gctgacttct ttggcaaaat
     1081 ttccttccag aagctttctt gaaatattga tgtcaaaaaa ttgtttaggt ctgacactaa
     1141 atggttcctg tgttttgtca atgagtccag tcttcatttt ttcatgtcgt ggacttataa
     1201 tttcacgttc aatacactgt agacgaatat taaaatcaca gtctccaact ggttgtgcat
     1261 tgaacttaat tatgtcatag atcaaatatc ttgggacggc ctgtccattt actttgtcaa
     1321 tgatcatttc cccatccaaa agagtatttg ataaatgcat gcgaagatct ttacgaaatg
     1381 gaaattccag atttgaaaca tgaaaaacgg aattatctct gtcaatcata aaaacttcat
     1441 ttgtgccatc aatcaacatc atgtaacgag tgccgtctgc tttccagctt actttataag
     1501 gcttctgctc taaaagtcta atattttgct tgtccatgga gacaggctgt gctccaggga
     1561 acccagaccc ttcccagcca cagaactgat gacatttctg ctgcacctct cctaacttgg
     1621 gttgagttgt tacttgagtt acacctttaa cagttatgcc ttccaagaaa atagctccca
     1681 gttttaaccg ctcttttctc cttttactaa aggatgcact tgaccctggt tctgagtcct
     1741 tttttccatc ttcatcctca tcttcttcat cctcatcttc aaaacaccaa tctggcaata
     1801 caggtggtgg tggtgcttct tctatatctc catagcgacg aaaaagttcc ttcaaataat
     1861 cacccttgta gattcctggt ggtctggctt gggcaaatgt agcaactgct gcttcaatac
     1921 tccaatccat tttttccacc aaaaaggcac atataaggaa accagtgcgg ttgaagccat
     1981 gggtgcagtg aacacctata agttcaggtg ggcttctttc attaaacctc tcacacaggc
     2041 gaatgaatgt ctcagtattc tcagtggtgg ggcattcgcc atgtccttta cactgaagtt
     2101 tgatatactt gattccttct ttttctatgt catttctgtc atagaacctt gaagtatttg
     2161 tcagatccac caacaagctc attttaacct tcaggctctt tagataattt gagagcatac
     2221 tgggatggaa ccggttttct tcagcaactt gactatcata tcttggcccc agcatcgtct
     2281 tcagaggtaa gaatcttcct gccaccggct gaccgcgccg cggacagttc agccaccgcg
     2341 gcgggatctt gttgtaagcg gcctccatgg ccataagcag gtcctcctct gagatcagct
     2401 tctgcattga tgccatggtg gtaccaagct tgggtctccc tatagtgagt cgtattaatt
     2461 tcgataagcc agtaagcagt gggttctcta gttagccaga gagctctgct tatatagacc
     2521 tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt
     2581 ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga
     2641 cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc
     2701 atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
     2761 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
     2821 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
     2881 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     2941 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     3001 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat
     3061 taataactag tcaataatca atgtcaacgc gtatatctgg cccgtacatc gcgaagcagc
     3121 gcaaaacgcc taaccctaag cagattcttc atgcaattgt cggtcaagcc ttgccttgtt
     3181 gtagcttaaa ttttgctcgc gcactactca gcgacctcca acacacaagc agggagcaga
     3241 tactggctta actatgcggc atcagagcag attgtactga gagtgcacca taggggatcg
     3301 ggagatctcc cgatccgtcg acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3361 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3421 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3481 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3541 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3601 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3661 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3721 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3781 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3841 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3901 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3961 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4021 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4081 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4141 ttgctgataa atctggagcc tccaagggtg ggtctcgcgg tatcattgca gcactggggc
     4201 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4261 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4321 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     4381 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4441 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     4501 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4561 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4621 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4681 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4741 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4801 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4861 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4921 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4981 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5041 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5101 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5161 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5221 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     5281 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     5341 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     5401 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     5461 caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac ggtatacaga
     5521 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     5581 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     5641 acaagttggg gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatccagc
     5701 cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg gtggaatcga
     5761 aatctcgtga tggcaggttg ggcgtcgctt ggtcggtcat ttcgaacccc agagtcccgc
     5821 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
     5881 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
     5941 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
     6001 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
     6061 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
     6121 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
     6181 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
     6241 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
     6301 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
     6361 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
     6421 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
     6481 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
     6541 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
     6601 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc cgaaaatgga
     6661 tatacaagct cccgggagct ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta
     6721 cttctggaat agctcagagg cagaggcggc ctcggcctct gcataaataa aaaaaattag
     6781 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
     6841 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
     6901 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
     6961 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
     7021 aattaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     7081 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     7141 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     7201 tatcaggg
//
