LOCUS       pcDNA3                  5789 bp    DNA     circular     03-MAY-2019
Description Mammalian expression vector with the CMV promoter. The MCS is in the forward (+) orientation.
Comments The cloned gene must include a Kozak sequence and start codon.
FEATURES             Location/Qualifiers
     promoter        615..818
                     /dnas_title="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        235..614
                     /dnas_title="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     CDS             complement(join(4793..5584,5585..5653))
                     /dnas_title="AmpR"
                     /product="%CE%B2-lactamase"
                     /transl_table=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /codon_start=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     CDS             2497..3291
                     /dnas_title="NeoR/KanR"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and
                     G418 (Geneticin)"
                     /codon_start=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     rep_origin      complement(4037..4622)
                     /dnas_title="ori"
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     rep_origin      1659..2087
                     /dnas_title="f1 ori"
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        2101..2430
                     /dnas_title="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     polyA_signal    1389..1613
                     /dnas_title="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2281..2416
                     /dnas_title="SV40 ori"
                     /note="SV40 origin of replication"
     polyA_signal    3465..3586
                     /dnas_title="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(5654..5758)
                     /dnas_title="AmpR promoter"
                     /gene="bla"
     promoter        863..881
                     /dnas_title="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             923..1345
                     /note="Granulin4+linker5"
                     /translation="MWTLVSWVALTAGLVAGSAWSHPQFEKGGGSGGGSGGSAWSHPQ
                     FEKGASDYKDDDDKDVKCDMEVSCPDGYTCCRLQSGAWGCCPFTQAVCCEDHIHCCPA
                     GFTCDTQKGTCEQGPHQVPWMEKAPAHLSLPDPQALKR"
     misc_feature    311..614
                     /dnas_title="CMV enhancer"
                     /note="Human cytomegalovirus (CMV) immediate early
                     enhancer"
                     /note="Human cytomegalovirus (CMV) immediate early
                     enhancer"
     regulatory      616..818
                     /dnas_title="CMV promoter"
                     /regulatory_class="promoter"
                     /note="Human cytomegalovirus (CMV) immediate early
                     promoter"
                     /note="Human cytomegalovirus (CMV) immediate early
                     promoter"
     regulatory      863..880
                     /dnas_title="T7 promoter"
                     /regulatory_class="promoter"
                     /note="bacteriophage T7 RNA polymerase promoter"
                     /note="bacteriophage T7 RNA polymerase promoter"
     misc_feature    1082..1096
                     /dnas_title="EK"
                     /note="several sequences are used to encode this tag"
                     /note="several sequences are used to encode this tag"
     regulatory      1389..1596
                     /dnas_title="BGH polyA signal"
                     /regulatory_class="polyA_signal_sequence"
                     /note="BGH polyadenylation sequence"
     rep_origin      complement(1659..2039)
                     /dnas_title="f1 ori"
                     /note="both + and - origins are located in the intergenic
                     region (IG)"
                     /note="both + and - origins are located in the intergenic
                     region (IG)"
     regulatory      3521..3595
                     /dnas_title="SV40 early polyA signal"
                     /regulatory_class="polyA_signal_sequence"
                     /note="the early polyadenylation signal (EPA) and the
                     late polyadenylation signal (LPA) are on opposite strands
                     of the same region"
                     /note="the early polyadenylation signal (EPA) and the
                     late polyadenylation signal (LPA) are on opposite strands
                     of the same region"
     CDS             complement(3625..3639)
                     /dnas_title="lacZa part A"
                     /gene="lacZ alpha"
                     /codon_start=1
                     /transl_table=11
                     /product="Beta-galactosidase alpha peptide"
                     /translation="MTMIT"
     regulatory      complement(3657..3677)
                     /dnas_title="lac operator"
                     /regulatory_class="transcriptional_cis_regulatory_region"
                     /note="Lac repressor binding site"
                     /note="Lac repressor binding site"
     regulatory      complement(3683..3713)
                     /dnas_title="lac promoter"
                     /regulatory_class="promoter"
                     /note="lac promoter"
     regulatory      complement(3724..3748)
                     /dnas_title="CAP binding site"
                     /regulatory_class="transcriptional_cis_regulatory_region"
                     /note="CAP binding site"
     regulatory      complement(5688..5723)
                     /dnas_title="AmpR promoter"
                     /regulatory_class="promoter"
                     /note="includes -35, -10, and transcription start site"
                     /note="includes -35, -10, and transcription start site"
     CDS             complement(4793..5587)
                     /dnas_title="bla(M)"
                     /gene="bla"
                     /gene_synonym="amp"
                     /codon_start=1
                     /transl_table=11
                     /product="Beta-lactamase TEM"
                     /EC_number="3.5.2.6"
                     /note="lacks the N-terminal 23 amino acid periplasmic
                     secretory signal sequence"
                     /translation="MDPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPM
                     MSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAA
                     ITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPV
                     AMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGS
                     RGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     CDS             complement(4793..5653)
                     /dnas_title="AmpR"
                     /gene="bla"
                     /gene_synonym="amp"
                     /codon_start=1
                     /transl_table=11
                     /product="Beta-lactamase TEM"
                     /EC_number="3.5.2.6"
                     /db_xref="RefSeq:WP_000027057.1"
                     /note="Penicillinase; ampicillin resistance gene"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     regulatory      2114..2421
                     /dnas_title="SV40 promoter"
                     /regulatory_class="promoter"
                     /note="overlaps the SV40 origin of replication"
                     /note="Simian virus 40 early promoter"
                     /note="includes 72-bp direct repeat (transcriptional
                     enhancer), 21-bp repeats containing six GC boxes, and the
                     TATA box"
                     /note="overlaps the SV40 origin of replication"
     rep_origin      complement(4027..4648)
                     /dnas_title="pMB1 ori"
                     /note="C171T is a pUC mutation; none of the other
                     observed variants are described in the literature"
                     /note="C171T is a pUC mutation; none of the other
                     observed variants are described in the literature"
     rep_origin      2333..2396
                     /dnas_title="SV40 ori"
                     /note="this core origin is both necessary and sufficient
                     for replication"
                     /note="this core origin is both necessary and sufficient
                     for replication"
     misc_feature    974..1003
                     /note="Strep-tag II"
     misc_feature    1034..1063
                     /note="Strep-tag II"
     misc_feature    1074..1096
                     /note="FLAG tag"
     source          1..5789
                     /dnas_title="pcDNA3.1(+)_GRN4-linker5"
ORIGIN      
        1 GACGGATCGG GAGATCTCCC GATCCCCTAT GGTGCACTCT CAGTACAATC TGCTCTGATG
       61 CCGCATAGTT AAGCCAGTAT CTGCTCCCTG CTTGTGTGTT GGAGGTCGCT GAGTAGTGCG
      121 CGAGCAAAAT TTAAGCTACA ACAAGGCAAG GCTTGACCGA CAATTGCATG AAGAATCTGC
      181 TTAGGGTTAG GCGTTTTGCG CTGCTTCGCG ATGTACGGGC CAGATATACG CGTTGACATT
      241 GATTATTGAC TAGTTATTAA TAGTAATCAA TTACGGGGTC ATTAGTTCAT AGCCCATATA
      301 TGGAGTTCCG CGTTACATAA CTTACGGTAA ATGGCCCGCC TGGCTGACCG CCCAACGACC
      361 CCCGCCCATT GACGTCAATA ATGACGTATG TTCCCATAGT AACGCCAATA GGGACTTTCC
      421 ATTGACGTCA ATGGGTGGAG TATTTACGGT AAACTGCCCA CTTGGCAGTA CATCAAGTGT
      481 ATCATATGCC AAGTACGCCC CCTATTGACG TCAATGACGG TAAATGGCCC GCCTGGCATT
      541 ATGCCCAGTA CATGACCTTA TGGGACTTTC CTACTTGGCA GTACATCTAC GTATTAGTCA
      601 TCGCTATTAC CATGGTGATG CGGTTTTGGC AGTACATCAA TGGGCGTGGA TAGCGGTTTG
      661 ACTCACGGGG ATTTCCAAGT CTCCACCCCA TTGACGTCAA TGGGAGTTTG TTTTGGCACC
      721 AAAATCAACG GGACTTTCCA AAATGTCGTA ACAACTCCGC CCCATTGACG CAAATGGGCG
      781 GTAGGCGTGT ACGGTGGGAG GTCTATATAA GCAGAGCTCT CTGGCTAACT AGAGAACCCA
      841 CTGCTTACTG GCTTATCGAA ATTAATACGA CTCACTATAG GGAGACCCAA GCTGGCTAGC
      901 GTTTAAACTT AAGCTTGCCA CCATGTGGAC TCTGGTGTCC TGGGTCGCAC TGACCGCAGG
      961 ACTGGTGGCT GGAAGCGCAT GGTCCCACCC ACAGTTCGAG AAGGGAGGAG GATCTGGAGG
     1021 AGGATCTGGA GGGAGTGCAT GGTCACATCC TCAGTTTGAG AAGGGAGCCA GTGACTACAA
     1081 AGACGATGAC GATAAGGACG TGAAATGCGA TATGGAAGTC AGCTGTCCAG ATGGGTATAC
     1141 ATGCTGTCGA CTGCAGTCCG GAGCATGGGG ATGCTGTCCT TTCACTCAGG CCGTGTGCTG
     1201 TGAAGACCAC ATCCATTGCT GTCCAGCTGG CTTTACCTGC GATACACAGA AAGGCACCTG
     1261 TGAGCAGGGG CCACACCAGG TCCCCTGGAT GGAAAAGGCA CCAGCTCATC TGAGCCTGCC
     1321 AGACCCTCAG GCCCTGAAAA GATGACTCGA GTCTAGAGGG CCCGTTTAAA CCCGCTGATC
     1381 AGCCTCGACT GTGCCTTCTA GTTGCCAGCC ATCTGTTGTT TGCCCCTCCC CCGTGCCTTC
     1441 CTTGACCCTG GAAGGTGCCA CTCCCACTGT CCTTTCCTAA TAAAATGAGG AAATTGCATC
     1501 GCATTGTCTG AGTAGGTGTC ATTCTATTCT GGGGGGTGGG GTGGGGCAGG ACAGCAAGGG
     1561 GGAGGATTGG GAAGACAATA GCAGGCATGC TGGGGATGCG GTGGGCTCTA TGGCTTCTGA
     1621 GGCGGAAAGA ACCAGCTGGG GCTCTAGGGG GTATCCCCAC GCGCCCTGTA GCGGCGCATT
     1681 AAGCGCGGCG GGTGTGGTGG TTACGCGCAG CGTGACCGCT ACACTTGCCA GCGCCCTAGC
     1741 GCCCGCTCCT TTCGCTTTCT TCCCTTCCTT TCTCGCCACG TTCGCCGGCT TTCCCCGTCA
     1801 AGCTCTAAAT CGGGGGCTCC CTTTAGGGTT CCGATTTAGT GCTTTACGGC ACCTCGACCC
     1861 CAAAAAACTT GATTAGGGTG ATGGTTCACG TAGTGGGCCA TCGCCCTGAT AGACGGTTTT
     1921 TCGCCCTTTG ACGTTGGAGT CCACGTTCTT TAATAGTGGA CTCTTGTTCC AAACTGGAAC
     1981 AACACTCAAC CCTATCTCGG TCTATTCTTT TGATTTATAA GGGATTTTGC CGATTTCGGC
     2041 CTATTGGTTA AAAAATGAGC TGATTTAACA AAAATTTAAC GCGAATTAAT TCTGTGGAAT
     2101 GTGTGTCAGT TAGGGTGTGG AAAGTCCCCA GGCTCCCCAG CAGGCAGAAG TATGCAAAGC
     2161 ATGCATCTCA ATTAGTCAGC AACCAGGTGT GGAAAGTCCC CAGGCTCCCC AGCAGGCAGA
     2221 AGTATGCAAA GCATGCATCT CAATTAGTCA GCAACCATAG TCCCGCCCCT AACTCCGCCC
     2281 ATCCCGCCCC TAACTCCGCC CAGTTCCGCC CATTCTCCGC CCCATGGCTG ACTAATTTTT
     2341 TTTATTTATG CAGAGGCCGA GGCCGCCTCT GCCTCTGAGC TATTCCAGAA GTAGTGAGGA
     2401 GGCTTTTTTG GAGGCCTAGG CTTTTGCAAA AAGCTCCCGG GAGCTTGTAT ATCCATTTTC
     2461 GGATCTGATC AAGAGACAGG ATGAGGATCG TTTCGCATGA TTGAACAAGA TGGATTGCAC
     2521 GCAGGTTCTC CGGCCGCTTG GGTGGAGAGG CTATTCGGCT ATGACTGGGC ACAACAGACA
     2581 ATCGGCTGCT CTGATGCCGC CGTGTTCCGG CTGTCAGCGC AGGGGCGCCC GGTTCTTTTT
     2641 GTCAAGACCG ACCTGTCCGG TGCCCTGAAT GAACTGCAGG ACGAGGCAGC GCGGCTATCG
     2701 TGGCTGGCCA CGACGGGCGT TCCTTGCGCA GCTGTGCTCG ACGTTGTCAC TGAAGCGGGA
     2761 AGGGACTGGC TGCTATTGGG CGAAGTGCCG GGGCAGGATC TCCTGTCATC TCACCTTGCT
     2821 CCTGCCGAGA AAGTATCCAT CATGGCTGAT GCAATGCGGC GGCTGCATAC GCTTGATCCG
     2881 GCTACCTGCC CATTCGACCA CCAAGCGAAA CATCGCATCG AGCGAGCACG TACTCGGATG
     2941 GAAGCCGGTC TTGTCGATCA GGATGATCTG GACGAAGAGC ATCAGGGGCT CGCGCCAGCC
     3001 GAACTGTTCG CCAGGCTCAA GGCGCGCATG CCCGACGGCG AGGATCTCGT CGTGACCCAT
     3061 GGCGATGCCT GCTTGCCGAA TATCATGGTG GAAAATGGCC GCTTTTCTGG ATTCATCGAC
     3121 TGTGGCCGGC TGGGTGTGGC GGACCGCTAT CAGGACATAG CGTTGGCTAC CCGTGATATT
     3181 GCTGAAGAGC TTGGCGGCGA ATGGGCTGAC CGCTTCCTCG TGCTTTACGG TATCGCCGCT
     3241 CCCGATTCGC AGCGCATCGC CTTCTATCGC CTTCTTGACG AGTTCTTCTG AGCGGGACTC
     3301 TGGGGTTCGA AATGACCGAC CAAGCGACGC CCAACCTGCC ATCACGAGAT TTCGATTCCA
     3361 CCGCCGCCTT CTATGAAAGG TTGGGCTTCG GAATCGTTTT CCGGGACGCC GGCTGGATGA
     3421 TCCTCCAGCG CGGGGATCTC ATGCTGGAGT TCTTCGCCCA CCCCAACTTG TTTATTGCAG
     3481 CTTATAATGG TTACAAATAA AGCAATAGCA TCACAAATTT CACAAATAAA GCATTTTTTT
     3541 CACTGCATTC TAGTTGTGGT TTGTCCAAAC TCATCAATGT ATCTTATCAT GTCTGTATAC
     3601 CGTCGACCTC TAGCTAGAGC TTGGCGTAAT CATGGTCATA GCTGTTTCCT GTGTGAAATT
     3661 GTTATCCGCT CACAATTCCA CACAACATAC GAGCCGGAAG CATAAAGTGT AAAGCCTGGG
     3721 GTGCCTAATG AGTGAGCTAA CTCACATTAA TTGCGTTGCG CTCACTGCCC GCTTTCCAGT
     3781 CGGGAAACCT GTCGTGCCAG CTGCATTAAT GAATCGGCCA ACGCGCGGGG AGAGGCGGTT
     3841 TGCGTATTGG GCGCTCTTCC GCTTCCTCGC TCACTGACTC GCTGCGCTCG GTCGTTCGGC
     3901 TGCGGCGAGC GGTATCAGCT CACTCAAAGG CGGTAATACG GTTATCCACA GAATCAGGGG
     3961 ATAACGCAGG AAAGAACATG TGAGCAAAAG GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG
     4021 CCGCGTTGCT GGCGTTTTTC CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC
     4081 GCTCAAGTCA GAGGTGGCGA AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG
     4141 GAAGCTCCCT CGTGCGCTCT CCTGTTCCGA CCCTGCCGCT TACCGGATAC CTGTCCGCCT
     4201 TTCTCCCTTC GGGAAGCGTG GCGCTTTCTC ATAGCTCACG CTGTAGGTAT CTCAGTTCGG
     4261 TGTAGGTCGT TCGCTCCAAG CTGGGCTGTG TGCACGAACC CCCCGTTCAG CCCGACCGCT
     4321 GCGCCTTATC CGGTAACTAT CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC
     4381 TGGCAGCAGC CACTGGTAAC AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT
     4441 TCTTGAAGTG GTGGCCTAAC TACGGCTACA CTAGAAGAAC AGTATTTGGT ATCTGCGCTC
     4501 TGCTGAAGCC AGTTACCTTC GGAAAAAGAG TTGGTAGCTC TTGATCCGGC AAACAAACCA
     4561 CCGCTGGTAG CGGTTTTTTT GTTTGCAAGC AGCAGATTAC GCGCAGAAAA AAAGGATCTC
     4621 AAGAAGATCC TTTGATCTTT TCTACGGGGT CTGACGCTCA GTGGAACGAA AACTCACGTT
     4681 AAGGGATTTT GGTCATGAGA TTATCAAAAA GGATCTTCAC CTAGATCCTT TTAAATTAAA
     4741 AATGAAGTTT TAAATCAATC TAAAGTATAT ATGAGTAAAC TTGGTCTGAC AGTTACCAAT
     4801 GCTTAATCAG TGAGGCACCT ATCTCAGCGA TCTGTCTATT TCGTTCATCC ATAGTTGCCT
     4861 GACTCCCCGT CGTGTAGATA ACTACGATAC GGGAGGGCTT ACCATCTGGC CCCAGTGCTG
     4921 CAATGATACC GCGAGACCCA CGCTCACCGG CTCCAGATTT ATCAGCAATA AACCAGCCAG
     4981 CCGGAAGGGC CGAGCGCAGA AGTGGTCCTG CAACTTTATC CGCCTCCATC CAGTCTATTA
     5041 ATTGTTGCCG GGAAGCTAGA GTAAGTAGTT CGCCAGTTAA TAGTTTGCGC AACGTTGTTG
     5101 CCATTGCTAC AGGCATCGTG GTGTCACGCT CGTCGTTTGG TATGGCTTCA TTCAGCTCCG
     5161 GTTCCCAACG ATCAAGGCGA GTTACATGAT CCCCCATGTT GTGCAAAAAA GCGGTTAGCT
     5221 CCTTCGGTCC TCCGATCGTT GTCAGAAGTA AGTTGGCCGC AGTGTTATCA CTCATGGTTA
     5281 TGGCAGCACT GCATAATTCT CTTACTGTCA TGCCATCCGT AAGATGCTTT TCTGTGACTG
     5341 GTGAGTACTC AACCAAGTCA TTCTGAGAAT AGTGTATGCG GCGACCGAGT TGCTCTTGCC
     5401 CGGCGTCAAT ACGGGATAAT ACCGCGCCAC ATAGCAGAAC TTTAAAAGTG CTCATCATTG
     5461 GAAAACGTTC TTCGGGGCGA AAACTCTCAA GGATCTTACC GCTGTTGAGA TCCAGTTCGA
     5521 TGTAACCCAC TCGTGCACCC AACTGATCTT CAGCATCTTT TACTTTCACC AGCGTTTCTG
     5581 GGTGAGCAAA AACAGGAAGG CAAAATGCCG CAAAAAAGGG AATAAGGGCG ACACGGAAAT
     5641 GTTGAATACT CATACTCTTC CTTTTTCAAT ATTATTGAAG CATTTATCAG GGTTATTGTC
     5701 TCATGAGCGG ATACATATTT GAATGTATTT AGAAAAATAA ACAAATAGGG GTTCCGCGCA
     5761 CATTTCCCCG AAAAGTGCCA CCTGACGTC
//

