LOCUS       pcDNA3_N3FLAG_MY        7401 bp ds-DNA     circular     18-DEC-2017
DEFINITION  Mammalian expression vector with the CMV promoter and a
            neomycin-resistance marker.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5446)
  AUTHORS   Invitrogen (Life Technologies)
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Dec 18, 2017 from SnapGene Viewer 3.3.4
            http://www.snapgene.com
COMMENT     The cloned gene must include a Kozak sequence and start codon.
COMMENT     
COMMENT     ApEinfo:methylated:1
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label=CMV enhancer
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label=CMV promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label=T7 promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    916..918
                     /label=3*Flag
                     /ApEinfo_fwdcolor=#cc66ff
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    join(889..912,2926..2949)
                     /note="multiple cloning site"
                     /label=MCS
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    940..1005
                     /label=3*Flag(1)
                     /ApEinfo_label=3*Flag
                     /ApEinfo_fwdcolor=#cc66ff
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        complement(2953..2971)
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /label=SP6 promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    919..939
                     /label=NLS
                     /ApEinfo_fwdcolor=#00ff00
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     polyA_signal    2997..3221
                     /note="bovine growth hormone polyadenylation signal"
                     /label=bGH poly(A) signal
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    1012..1017
                     /label=EcoRI
                     /ApEinfo_fwdcolor=#ff6fcf
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      3267..3695
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label=f1 ori
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     misc_feature    1018..2925
                     /label=MYB-FL
                     /ApEinfo_fwdcolor=#ff6666
                     /ApEinfo_revcolor=green
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        3709..4039
                     /note="SV40 enhancer and early promoter"
                     /label=SV40 promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      3890..4025
                     /note="SV40 origin of replication"
                     /label=SV40 ori
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             4106..4900
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin)"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     P
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     S
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     Q
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     A LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
                     /label=NeoR/KanR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     polyA_signal    5074..5195
                     /note="SV40 polyadenylation signal"
                     /label=SV40 poly(A) signal
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     primer_bind     complement(5244..5260)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label=M13 rev
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     rep_origin      complement(5646..6234)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label=ori
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     CDS             complement(6405..7265)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     I
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     S
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     W
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     A
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     S LIKHW"
                     /label=AmpR
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
     promoter        complement(7266..7370)
                     /gene="bla"
                     /label=AmpR promoter
                     /ApEinfo_fwdcolor=pink
                     /ApEinfo_revcolor=pink
                     /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0}
                     width 5 offset 0
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc
      901 gagctcggat ccGCCATGCC AAAGAAGAAG CGTAAGGTAG ATTACAAAGA CCATGACGGT
      961 GATTATAAAG ATCATGATAT CGATTACAAG GATGACGATG ACAAGGGAGG CgaattcGCC
     1021 CGGAGACCCC GACACAGCAT CTACAGTAGC GATGAAGATG ATGAAGACAT TGAGATGTGT
     1081 GACCATGACT ACGATGGGCT GCTGCCCAAA TCTGGAAAGC GTCACTTGGG GAAAACTAGG
     1141 TGGACAAGGG AAGAGGATGA GAAGCTGAAG AAGCTGGTGG AACAGAACGG AACAGACGAC
     1201 TGGAAAGTCA TTGCCAATTA TCTGCCCAAC CGGACAGATG TGCAGTGCCA ACACCGGTGG
     1261 CAGAAAGTGC TGAACCCTGA ACTCATCAAA GGTCCCTGGA CCAAAGAAGA AGATCAGAGA
     1321 GTCATAGAGC TTGTCCAGAA ATATGGTCCG AAGCGTTGGT CTGTTATTGC CAAGCACTTA
     1381 AAAGGGAGAA TTGGAAAGCA GTGTCGGGAG AGGTGGCACA ACCATTTGAA TCCAGAAGTT
     1441 AAGAAAACCT CCTGGACAGA AGAGGAGGAC AGAATCATTT ACCAGGCACA CAAGCGTCTG
     1501 GGGAACAGAT GGGCAGAGAT CGCAAAGCTG CTGCCCGGAC GGACTGATAA TGCTATCAAG
     1561 AACCACTGGA ATTCCACCAT GCGTCGCAAG GTGGAACAGG AAGGCTACCT GCAGGAGCCT
     1621 TCCAAAGCCA GCCAGACGCC AGTGGCCACG AGCTTCCAGA AGAACAATCA TTTGATGGGG
     1681 TTTGGGCATG CCTCACCTCC ATCTCAGCTC TCTCCAAGTG GCCAGTCCTC CGTCAACAGC
     1741 GAATATCCCT ATTACCACAT CGCCGAAGCA CAAAACATCT CCAGTCACGT TCCCTATCCT
     1801 GTCGCATTGC ATGTTAATAT AGTCAACGTC CCTCAGCCGG CTGCGGCAGC CATCCAGAGA
     1861 CACTATAACG ACGAAGACCC TGAGAAGGAA AAGCGAATAA AGGAGCTGGA GTTGCTCCTG
     1921 ATGTCAACAG AGAACGAGCT GAAGGGACAG CAGGCATTAC CAACACAGAA CCACACTTGC
     1981 AGCTACCCCG GGTGGCACAG CACCTCCATT GTGGACCAGA CCAGACCTCA TGGGGATAGT
     2041 GCACCTGTTT CCTGTTTGGG AGAACACCAT GCCACCCCAT CTCTGCCTGC AGATCCCGGC
     2101 TCCCTACCTG AAGAAAGTGC CTCACCAGCA AGGTGCATGA TCGTCCACCA GGGCACCATT
     2161 CTGGACAATG TTAAGAACCT CTTAGAATTT GCAGAAACAC TCCAGTTTAT AGATTCTTTC
     2221 TTGAACACTT CCAGCAACCA TGAAAGCTCG GGCTTAGATG CACCTACCTT ACCCTCCACT
     2281 CCTCTCATTG GTCACAAACT GACACCATGT CGAGACCAGA CTGTGAAAAC CCAGAAGGAA
     2341 AATTCCATCT TTAGAACTCC AGCTATCAAA AGGTCAATCC TCGAAAGCTC TCCTCGAACT
     2401 CCCACACCAT TCAAACATGC CCTTGCAGCT CAAGAAATTA AATACGGTCC CCTGAAGATG
     2461 CTACCTCAGA CCCCCTCCCA TGCAGTGGAG GACCTACAAG ATGTGATTAA GCAGGAATCG
     2521 GATGAATCTG GAATTGTGGC TGAGTTTCAA GAGAGTGGAC CACCGTTACT GAAAAAAATC
     2581 AAGCAGGAGG TGGAGTCGCC AACTGAGAAA TCGGGAAACT TCTTCTGCTC AAACCACTGG
     2641 GCAGAGAACA GCCTGAGCAC CCAGCTGTTC TCGCAGGCGT CTCCTGTGGC AGATGCCCCA
     2701 AATATTCTTA CAAGCTCTGT TTTAATGACA CCTGTATCAG AAGATGAAGA CAATGTCCTC
     2761 AAAGCCTTTA CCGTACCTAA GAACAGGCCC CTGGTGGGTC CCTTGCAGCC ATGCAGTGGT
     2821 GCCTGGGAGC CAGCATCCTG TGGGAAGACA GAGGACCAGA TGACGGCCTC CGGTCCGGCT
     2881 CGGAAATACG TGAACGCGTT CTCAGCTCGA ACTCTGGTCA TGTGActcga gcatgcatct
     2941 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
     3001 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3061 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3121 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3181 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     3241 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     3301 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     3361 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3421 gggcatccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3481 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3541 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     3601 tatctcggtc tattcttttg atttataagg gattttgggg atttcggcct attggttaaa
     3661 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     3721 gggtgtggaa agtccccagg ctccccaggc aggcagaagt atgcaaagca tgcatctcaa
     3781 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3841 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3901 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3961 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     4021 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     4081 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4141 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4201 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4261 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     4321 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4381 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4441 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4501 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4561 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4621 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4681 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4741 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4801 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4861 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4921 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4981 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     5041 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     5101 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     5161 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     5221 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     5281 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     5341 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5401 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5461 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5521 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5581 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5641 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5701 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5761 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5821 ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5881 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5941 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6001 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6061 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     6121 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6181 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6241 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6301 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6361 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6421 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6481 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6541 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6601 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6661 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6721 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6781 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6841 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6901 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6961 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7021 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7081 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7141 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7201 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7261 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7321 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7381 cgaaaagtgc cacctgacgt c   
//
